Biological technologies (biotechnologies) provide controlled production healthy products for various spheres of human activity, based on the use of the catalytic potential of biological agents and systems of varying degrees of organization and complexity - microorganisms, viruses, plant and animal cells and tissues, as well as extracellular substances and cell components.

The development and transformation of biotechnology is driven by profound changes that have occurred in biology over the past 25-30 years. These events were based on new ideas in the field of molecular biology and molecular genetics. At the same time, it should be noted that the development and achievements of biotechnology are closely related to the body of knowledge not only of biological sciences, but also of many others.

The expansion of the practical sphere of biotechnology is also due to the socio-economic needs of society. Such urgent problems facing humanity on the threshold of the 21st century, such as shortage clean water and nutrients (especially protein), environmental pollution, lack of raw materials and energy resources, the need to obtain new, environmentally friendly materials, the development of new diagnostic and treatment tools, cannot be solved by traditional methods. Therefore, to ensure human life support, improve the quality of life and its duration, it is becoming increasingly necessary to master fundamentally new methods and technologies.

The development of scientific and technological progress, accompanied by an increase in the rate of material and energy resources, unfortunately, leads to an imbalance in biosphere processes. The water and air basins of cities are polluted, the reproductive function of the biosphere is reduced, and due to the accumulation of dead-end products of the technosphere, global circulation cycles of the biosphere are disrupted.

The rapid pace of modern scientific and technological progress of mankind was figuratively described by the Swiss engineer and philosopher Eichelberg: “It is believed that the age of mankind is 600,000 years. Let’s imagine the movement of humanity in the form of a 60 km marathon, which, starting somewhere, goes towards the center of one of our cities, as if towards the finish... Most of the distance runs along a very difficult path - through virgin forests, and we We don’t know anything about this, because only at the very end, at 58-59 km of running, we find, along with primitive tools, cave drawings as the first signs of culture, and only at the last kilometer do signs of agriculture appear.

200 m before the finish line, a road covered with stone slabs leads past Roman fortifications. 100 meters away, the runners are surrounded by medieval city buildings. There are 50 meters left before the finish line, where a man stands, watching the runners with intelligent and understanding eyes - this is Leonardo da Vinci. There are 10 m left. They begin in the light of torches and the poor lighting of oil lamps. But when throwing in the last 5 meters, a stunning miracle occurs: the light floods the night road, carts without draft animals rush past, cars rustle in the air, and the amazed runner is blinded by the light of the spotlights of photo and television cameras...”, i.e. in 1 m, the human genius makes a stunning leap in the field of scientific and technological progress. Continuing this image, we can add that as the runner approaches the finish line, thermonuclear fusion is tamed, spaceships are launched, and the genetic code is deciphered.

Biotechnology is the basis of scientific and technological progress and improving the quality of human life

Biotechnology as a field of knowledge and a dynamically developing industrial sector is designed to solve many key problems of our time, while ensuring the preservation of balance in the system of relationships “man - nature - society”, because biological technologies (biotechnologies), based on the use of the potential of living things, are by definition aimed at friendliness and harmony of a person with the world around him. Currently, biotechnology is divided into several most significant segments: these are “white”, “green”, “red”, “gray” and “blue” biotechnology.

“White” biotechnology includes industrial biotechnology, focused on the production of products previously produced by the chemical industry - alcohol, vitamins, amino acids, etc. (taking into account the requirements of resource conservation and environmental protection).

Green biotechnology covers an area of ​​relevance to agriculture. This is research and technology aimed at creating biotechnological methods and drugs to combat pests and pathogens. cultivated plants and domestic animals, creating biofertilizers, increasing plant productivity, including using genetic engineering methods.

Red (medical) biotechnology is the most significant area of ​​modern biotechnology. This is the production of diagnosticums using biotechnological methods and medicines using cellular and genetic engineering technologies (green vaccines, gene diagnostics, monoclonal antibodies, tissue engineering constructs and products, etc.).

Gray biotechnology develops technologies and drugs to protect the environment; these are soil reclamation, wastewater and gaseous emissions treatment, industrial waste disposal and toxicant degradation using biological agents and biological processes.

Blue biotechnology is mainly focused on the efficient use of ocean resources. First of all, this is the use of marine biota to obtain food, technical, biologically active and medicinal substances.

Modern biotechnology is one of the priority areas of the national economy of all developed countries. The way to increase the competitiveness of biotechnological products in sales markets is one of the main ones in the overall strategy for the development of biotechnology in industrialized countries. A stimulating factor is specially adopted government programs for the accelerated development of new areas of biotechnology.

State programs provide for the issuance of gratuitous loans to investors, long-term loans, and tax exemptions. As basic and targeted research becomes increasingly costly, many countries are seeking to move significant research beyond national borders.

As is known, the probability of success of R&D projects in general does not exceed 12-20%, about 60% of projects reach the stage of technical completion, 30% - commercial development, and only 12% are profitable.

Features of the development of research and commercialization of biological technologies in the USA, Japan, EU countries and Russia

USA. The leading position in biotechnology in terms of industrial production of biotechnological products, sales volumes, foreign trade turnover, allocations and scale of R&D is occupied by the United States, where great attention is paid to the development of this area. By 2003, over 198,300 people were employed in this sector.

Allocations to this sector of science and economics in the United States are significant and amount to over $20 billion. USA annually. Revenues of the US biotechnology industry increased from $8 billion. in 1992 to 39 billion dollars. in 2003

This industry is under close government attention. Thus, during the period of formation of the latest biotechnology and the emergence of its directions related to the manipulation of genetic material, in the mid-70s. last century, the US Congress paid great attention to the safety of genetic research. In 1977 alone, 25 special hearings were held and 16 bills were passed.

In the early 90s. The focus has shifted to developing measures to encourage the practical use of biotechnology for the production of new products. The development of biotechnology in the United States is associated with the solution of many key problems: energy, raw materials, food and environmental issues.

Among biotechnological areas close to practical implementation or at the stage of industrial development, the following:
- bioconversion of solar energy;
- the use of microorganisms to increase oil yield and leaching of non-ferrous and rare metals;
- designing strains that can replace expensive inorganic catalysts and change synthesis conditions to obtain fundamentally new compounds;
- the use of bacterial plant growth stimulants, changing the genotype of cereals and their adaptation to ripening in extreme conditions (without plowing, watering and fertilizers);
- directed biosynthesis for the effective production of target products (amino acids, enzymes, vitamins, antibiotics, food additives, pharmacological drugs;
- obtaining new diagnostic and therapeutic drugs based on cellular and genetic engineering methods.

The role of the US leader is due to the high allocations of government and private capital for basic and applied research. The National Science Foundation (NSF), the Departments of Health and Human Services, Agriculture, Energy, Chemicals and Food Industry, Defense, National Aeronautics and Space Administration (NASA), Interior. Allocations are allocated on a program-target basis, i.e. Research projects are subsidized and contracted.

At the same time, large industrial companies establish business relationships with universities and research centers. This contributes to the formation of complexes in one area or another, ranging from basic research before serial production of the product and delivery to the market. This “participation system” provides for the formation of specialized funds with appropriate expert councils and the attraction of the most qualified personnel.

When selecting projects with high commercial impact, it has become advantageous to use the so-called “constraint analysis.” This allows you to significantly reduce the project implementation time (on average from 7-10 to 2-4 years) and increase the probability of success to 80%. The concept of “specified limitations” includes the potential for successful sale of the product and making a profit, increasing annual production, competitiveness of the product, potential risk from a sales perspective, the possibility of restructuring production taking into account new achievements, etc.

Annual total US government spending on genetic engineering and biotechnology research amounts to billions of dollars. Investments from private companies significantly exceed these figures. Several billion dollars are allocated annually for the creation of diagnostic and anticancer drugs alone. These are mainly the following areas: methods of DNA recombination, production of hybrids, production and use of monoclonal antibodies, tissue and cell culture.

In the United States, it has become common for companies not previously associated with biotechnology to begin acquiring stakes in existing companies and building their own biotechnology enterprises (Table 1.1). This, for example, is the practice of such chemical giants as Philips Petrolium, Monsanto, Dow Chemical. About 250 chemical companies currently have interests in biotechnology. Thus, the giant of the US chemical industry, the De Pont company, has several biotechnological complexes worth 85-150 thousand dollars. with a staff of 700-1,000 people.

Similar complexes have been created within the Monsanto structure; moreover, currently up to 75% of the budget (over $750 million) is allocated to the field of biotechnology. The focus of these companies is the production of genetically engineered growth hormone, as well as a number of genetically engineered drugs for veterinary medicine and pharmacology. In addition, firms, together with university research centers, sign contracts for joint R&D.

Table 1.1. The largest US concerns and pharmaceutical companies producing medical biotechnological drugs

There is an opinion that all the necessary conditions for the formation and development of biotechnology in the United States have been prepared by the venture business. For large firms and companies, venture business is a well-established technique that allows them to obtain new developments in a shorter period of time, attracting small firms and small teams for this, rather than doing it on their own.

For example, in the 80s. General Electric, with the help of small firms, began to master the production of biologically active compounds; in 1981 alone, its risk allocations in biotechnology amounted to $3 million. Small firm risk-taking provides large companies and corporations with a mechanism for selecting economically viable innovations with strong commercial prospects.

ON THE. Voinov, T.G. Volova

Biotechnology as a science and sphere of production. Subject, goals and objectives of biotechnology, connection with fundamental disciplines.

Biotechnology is technological processes using biotechnological systems - living organisms and components of a living cell. Systems can be different - from microbes and bacteria to enzymes and genes. Biotechnology is a production based on the achievements of modern science: genetic engineering, physico-chemistry of enzymes, molecular diagnostics and molecular biology, selection genetics, microbiology, biochemistry, antibiotic chemistry.

In the field of production medicines Biotechnology is displacing traditional technologies and opening up fundamentally new opportunities. Biotechnological methods produce genetically engineered proteins (interferons, interleukins, insulin, vaccines against hepatitis, etc.), enzymes, diagnostic tools (test systems for drugs, medicinal substances, hormones, etc.), vitamins, antibiotics, biodegradable plastics, biocompatible materials.

Immune biotechnology, with the help of which single cells are recognized and isolated from mixtures, can be used not only directly in medicine for diagnosis and treatment, but also in scientific research, in the pharmacological, food and other industries, and can also be used to obtain drugs synthesized by cells the body's defense system.

Currently, biotechnology achievements are promising in the following industries:

In industry (food, pharmaceutical, chemical, oil and gas) - the use of biosynthesis and biotransformation of new substances based on strains of bacteria and yeast constructed by genetic engineering methods with specified properties based on microbiological synthesis;

In ecology - increasing the efficiency of eco-friendly plant protection, developing environmentally friendly cleaning technologies Wastewater, recycling of waste from the agro-industrial complex, design of ecosystems;

In the energy sector - the use of new sources of bioenergy obtained on the basis of microbiological synthesis and simulated photosynthetic processes, bioconversion of biomass into biogas;

In agriculture - development in the field of crop production of transgenic crops, biological plant protection products, bacterial fertilizers, microbiological methods, soil reclamation; in the field of animal husbandry - the creation of effective feed preparations from plant, microbial biomass and agricultural waste, animal reproduction based on embryogenetic methods;

In medicine - the development of medical biological products, monoclonal antibodies, diagnostics, vaccines, the development of immunobiotechnology in the direction of increasing the sensitivity and specificity of immunoassay for diseases of infectious and non-infectious nature.

Compared to chemical technology, biotechnology has the following main advantages:

The possibility of obtaining specific and unique natural substances, some of which (for example, proteins, DNA) cannot yet be obtained by chemical synthesis;

Carrying out biotechnological processes at relatively low temperatures and pressures;

Microorganisms have significantly higher rates of growth and accumulation of cell mass than other organisms. For example, with the help of microorganisms in a fermenter with a volume of 300 m 3, 1 t of protein can be produced per day (365 t/year). To produce the same amount of protein per year using cattle, you need to have a herd of 30,000 heads. If you use legumes, such as peas, to obtain such a rate of protein production, you will need to have a pea field with an area of ​​5400 hectares;

Cheap agricultural and industrial waste can be used as raw material in biotechnology processes;

Biotechnological processes, compared to chemical ones, are usually more environmentally friendly, have less harmful waste, and are close to natural processes occurring in nature;

As a rule, the technology and equipment in biotechnological production are simpler and cheaper.

The primary task facing biotechnology is the creation and development of production of drugs for medicine: interferons, insulins, hormones, antibiotics, vaccines, monoclonal antibodies and others, allowing for early diagnosis and treatment of cardiovascular, malignant, hereditary, infectious diseases, including viral diseases.

The concept of “biotechnology” is collective and covers such areas as fermentation technology, the use of biofactors using immobilized microorganisms or enzymes, genetic engineering, immune and protein technologies, technology using cell cultures of both animal and plant origin.

Biotechnology is a set of technological methods, including genetic engineering, using living organisms and biological processes for the production of medicines, or the science of the development and application of living systems, as well as non-living systems of biological origin within the framework of technological processes and industrial production.

Modern biotechnology is chemistry, where the change and transformation of substances occurs through biological processes. In intense competition, two chemistries are successfully developing: synthetic and biological.

1. Biological objects as a means of producing therapeutic, rehabilitation, preventive and diagnostic agents. Classification and general characteristics of biological objects.

Objects of biotechnology are viruses, bacteria, fungi - micromycetes and macromycetes, protozoal organisms, cells (tissues) of plants, animals and humans, some biogenic and functionally similar substances (for example, enzymes, prostaglandins, pectins, nucleic acids, etc.). Consequently, biotechnology objects can be represented by organized particles (viruses), cells (tissues) or their metabolites (primary, secondary). Even when a biomolecule is used as an object of biotechnology, its initial biosynthesis is carried out in most cases by the corresponding cells. In this regard, we can say that biotechnology objects relate either to microbes or to plant and animal organisms. In turn, the body can be figuratively characterized as a system of economical, complex, compact, self-regulating and, therefore, targeted biochemical production, steadily and actively proceeding with optimal maintenance of all necessary parameters. From this definition it follows that viruses are not organisms, but in terms of the content of molecules of heredity, adaptability, variability and some other properties they belong to representatives of living nature.

As can be seen from the diagram below, the objects of biotechnology are extremely diverse, their range extends from organized particles (viruses) to humans.

Currently, the majority of biotechnology objects are microbes belonging to three superkingdoms (non-nuclear, prenuclear, nuclear) and five kingdoms (viruses, bacteria, fungi, plants and animals). Moreover, the first two superkingdoms consist exclusively of microbes.

Microbes among plants are microscopic algae (Algae), and among animals - microscopic protozoa (Protozoa). Among eukaryotes, microbes include fungi and, with certain reservations, lichens, which are natural symbiotic associations of microscopic fungi and microalgae or fungi and cyanobacteria.

Acaryota - non-nuclear, Procaruota - prenuclear and Eucaruota - nuclear (from the Greek a - no, pro - to, eu - good, completely, caryota - core). The first includes organized particles - viruses and viroids, the second - bacteria, the third - all other organisms (fungi, algae, plants, animals).

Microorganisms form a huge number of secondary metabolites, many of which are also used, for example, antibiotics and other homeostasis correctors in mammalian cells.

Probiotics - preparations based on the biomass of certain types of microorganisms are used for dysbacteriosis to normalize the microflora of the gastrointestinal tract. Microorganisms are also needed in the production of vaccines. Finally, microbial cells can be converted using genetic engineering methods into producers of species-specific protein hormones for humans, protein factors of nonspecific immunity, etc.

Higher plants are the traditional and to date still the most extensive source of medicines. When using plants as biological objects, the main attention is focused on the issues of cultivating plant tissues on artificial media (callus and suspension cultures) and the new prospects that this opens up.

2. Macrobiological objects of animal origin. Man as a donor and object of immunization. Mammals, birds, reptiles, etc.

In recent years, due to the development of recombinant DNA technology, the importance of such a biological object as a person is rapidly increasing, although at first glance this seems paradoxical.

However, from the standpoint of biotechnology (using bioreactors), a person became a biological object only after realizing the possibility of cloning his DNA (more precisely, its exons) in microbial cells. Thanks to this approach, the shortage of raw materials for obtaining species-specific human proteins was eliminated.

Important in biotechnology are macro objects, which include various animals and birds. In the case of production of immune plasma, a person also acts as an object of immunization.

To obtain various vaccines, organs and tissues, including embryonic ones, of various animals and birds are used as objects for the propagation of viruses: It should be noted that the term "donor" V in this case denotes a biological object that supplies material for the production process of a medicinal product without harming its own life activity, and the term "donor"- a biological object from which the collection of material for the production of a medicinal product turns out to be incompatible with the continuation of life activity.

Of the embryonic tissues, chicken embryonic tissue is the most widely used. Of particular benefit are chicken embryos (according to availability) of ten to twelve days of age, used primarily for the reproduction of viruses and the subsequent production of viral vaccines. Chicken embryos were introduced into virological practice in 1931 by G. M. Woodruff and E. W. Goodpasture. Such embryos are also recommended for the detection, identification and determination of the infectious dose of viruses, for the production of antigenic preparations used in serological reactions.

Chicken eggs incubated at 38°C are ovoscoped (candled), “transparent” unfertilized specimens are rejected and fertilized ones are retained, in which the filled blood vessels of the chorioallantoic membrane and the movements of the embryos are clearly visible.

Infection of embryos can be carried out manually or automatically. The latter method is used in large-scale production, for example, of influenza vaccines. Material containing viruses is injected using a syringe (battery of syringes) into various parts of the embryo(s).

All stages of working with chicken embryos after ovoscopy are carried out under aseptic conditions. The material for infection can be a suspension of crushed brain tissue (in relation to the rabies virus), liver, spleen, kidneys (in relation to psittacosis chlamydia), etc. In order to decontaminate the viral material from bacteria or to prevent its bacterial contamination, appropriate antibiotics can be used , for example, penicillin with any aminoglycoside, about 150 IU of each per 1 ml of suspension of virus-containing material. To combat fungal infection of embryos, it is advisable to use some polyene antibiotics (nystatin, amphotericin B) or certain benzimidazole derivatives (for example, daktarin, etc.).

Most often, a suspension of viral material is injected into the allantoic cavity or, less commonly, onto the chorioallantoic membrane in an amount of 0.05-0.1 ml, piercing the disinfected shell (for example, with iodinated ethanol) to the calculated depth. After this, the hole is closed with molten paraffin and the embryos are placed in a thermostat, in which optimal temperature for virus reproduction, for example 36-37.5°C. The duration of incubation depends on the type and activity of the virus. Usually, after 2-4 days, a change in the membranes can be observed, followed by the death of the embryos. Infected embryos are monitored 1-2 times daily (ovoscoped, turned the other way). The dead embryos are then transferred to the viral material collection department. There they are disinfected, the allantoic fluid with the virus is sucked out and transferred to sterile containers. Inactivation of viruses at a certain temperature is usually carried out using formaldehyde, phenol or other substances. Using high-speed centrifugation or affinity chromatography (see), it is possible to obtain highly purified viral particles.

The collected viral material, which has passed appropriate control, is freeze-dried. The following indicators are subject to control: sterility, harmlessness and specific activity. With regard to sterility, they mean the absence of: a live homologous virus in a killed vaccine, bacteria and fungi. Safety and specific activity are assessed on animals and only after this the vaccine is allowed to be tested on volunteers or volunteers; after successful implementation After clinical testing, the vaccine is allowed to be used in widespread medical practice.

On chicken embryos, for example, live influenza vaccine. It is intended for intranasal administration (persons over 16 years old and children from 3 to 15 years old). The vaccine is a dried allantoic fluid taken from chicken embryos infected with the virus. The type of virus is selected according to the epidemiological situation and forecasts. Therefore, drugs can be produced in the form of a monovaccine or divaccine (for example, including viruses A2 and B) in ampoules with 20 and 8 vaccination doses for the relevant population groups. The dried mass in ampoules usually has a light yellow color, which remains even after the contents of the ampoule are dissolved in boiled, cooled water.

Live influenza vaccines for adults and children are also prepared for oral administration. Such vaccines are special vaccine strains, the reproduction of which occurred within 5-15 passages (no less and no more) on a culture of kidney tissue of chicken embryos. They are produced in dry form in bottles. When dissolved in water, the color changes from light yellow to reddish.

Other viral vaccines produced on chicken embryos include anti-mumps and yellow fever.

Other embryonic tissues include embryos of mice or other mammals, as well as aborted human fetuses.

Embryonic transplantable tissues are available after treatment with trypsin, since a large amount of intercellular substances (including non-protein nature) have not yet formed in such tissues. The cells are separated and, after the necessary treatments, they are cultured in special media in a monolayer or in a suspended state.

Tissues isolated from animals after birth are classified as mature. The older they are, the more difficult it is to cultivate them. However, once successfully grown, they then "flatten out" and are not much different from embryonic cells.

In addition to polio, specific prophylaxis with live vaccines is carried out for measles. Measles live dry vaccine are made from a vaccine strain, the reproduction of which was carried out on cell cultures of guinea pig kidneys or Japanese quail fibroblasts.

3. Biological objects of plant origin. Wild plants and plant cell cultures.

Plants are characterized by: the ability to photosynthesize, the presence of cellulose, and starch biosynthesis.

Algae are an important source of various polysaccharides and other biologically active substances. They reproduce vegetatively, asexually and sexually. As biological objects, they are not used enough, although, for example, kelp called seaweed is produced by industry in various countries. Agar-agar and alginates obtained from algae are well known.

Cells of higher plants. Higher plants (about 300,000 species) are differentiated multicellular, mainly terrestrial organisms. Of all the tissues, only meristematic ones are capable of division and at their expense all other tissues are formed. This is important for obtaining cells that must then be included in the biotechnological process.

Meristem cells that linger at the embryonic stage of development throughout the life of the plant are called initial; others gradually differentiate and turn into cells of various permanent tissues - terminal cells.

Depending on the topology in the plant, meristems are divided into apical, or apical (lat. arex - apex), lateral, or lateral (from lat. lateralis - lateral) and intermediate, or intercalary (from lat. Intercalaris - intermediate, inserted.

Totipotency- this is the property of plant somatic cells to fully realize their development potential up to the formation of a whole plant.

Any type of plant can, under appropriate conditions, produce an unorganized mass of dividing cells - callus (lat. callus - callus), especially under the inducing influence of plant hormones. Mass production of calli with further shoot regeneration is suitable for large-scale plant production. In general, callus is the main type of plant cell cultured on a nutrient medium. Callus tissue from any plant can be recultivated for a long time. In this case, the initial plants (including meristematic ones) are differentiated and despecialized, but are induced to divide, forming a primary callus.

In addition to growing calli, it is possible to cultivate cells of some plants in suspension cultures. Protoplasts of plant cells also appear to be important biological objects. The methods for obtaining them are fundamentally similar to the methods for obtaining bacterial and fungal protoplasts. Subsequent cell-based experiments with them are tempting due to the possible valuable results.

4. Biological objects - microorganisms. Main groups of obtained biologically active substances.

Objects of biotechnology are viruses, bacteria, fungi - micromycetes and macromycetes, protozoal organisms, cells (tissues) of plants, animals and humans, some biogenic and functionally similar substances (for example, enzymes, prostaglandins, lectins, nucleic acids, etc.). Consequently, biotechnology objects can be represented by organized particles (viruses), cells (tissues) or their metabolites (primary, secondary). Even when a biomolecule is used as an object of biotechnology, its initial biosynthesis is carried out in most cases by the corresponding cells. In this regard, we can say that biotechnology objects relate either to microbes or to plant and animal organisms. In turn, the body can be figuratively characterized as a system of economical, complex, compact, self-regulating and, therefore, targeted biochemical production, steadily and actively proceeding with optimal maintenance of all necessary parameters. From this definition it follows that viruses are not organisms, but in terms of the content of molecules of heredity, adaptability, variability and some other properties they belong to representatives of living nature.

Currently, the majority of biotechnology objects are microbes belonging to three superkingdoms (non-nuclear, prenuclear, nuclear) and five kingdoms (viruses, bacteria, fungi, plants and animals). Moreover, the first two superkingdoms consist exclusively of microbes.

The cells of fungi, algae, plants and animals have a real nucleus, delimited from the cytoplasm, and therefore they are classified as eukaryotes.

5. Biological objects - macromolecules with enzymatic activity. Use in biotechnological processes.

IN Lately a group of enzyme preparations has received a new direction of application - this is engineering enzymology, which is a branch of biotechnology where the biological object is an enzyme.

Organotherapy, i.e. treatment with organs and preparations from organs, tissues and secretions of animals, for a long time rested on deep empiricism and contradictory ideas, occupying a prominent place in medicine of all times and peoples. Only in the second half of the 19th century, as a result of the successes achieved by biological and organic chemistry and the development of experimental physiology, did organotherapy become scientifically based. This is associated with the name of the French physiologist Brown-Séquard. Particular attention was drawn to the work of Brown-Séquard associated with the introduction into the human body of extracts from the testes of a bull, which had a positive influence on performance and well-being.

The first official drugs (GF VII) were adrenaline, insulin, pituitrin, pepsin and pancreatin. Subsequently, as a result of extensive research conducted by Soviet endocrinologists and pharmacologists, it turned out to be possible to consistently expand the range of official and unofficial organ preparations.

However, some amino acids are obtained by chemical synthesis, for example, glycine, as well as D-, L-methionine, the D-isomer of which is low-toxic, therefore a medical preparation based on methionine contains D- and L-forms, although the drug is used in medicine abroad containing only the L-form of methionine. There, the racemic mixture of methionine is separated by bioconversion of the D-form into the L-form under the influence of special enzymes of living cells of microorganisms.

Immobilized enzyme preparations have a number of significant advantages when used for applied purposes compared to native precursors. Firstly, the heterogeneous catalyst is easy to separate from the reaction medium, which makes it possible to: a) stop the reaction at the right time; b) reuse the catalyst; c) obtain a product not contaminated with the enzyme. The latter is especially important in a number of food and pharmaceutical industries.

Secondly, the use of heterogeneous catalysts allows the enzymatic process to be carried out continuously, for example in flow columns, and the rate of the catalyzed reaction, as well as the product yield, to be controlled by changing the flow rate.

Thirdly, immobilization or modification of the enzyme contributes to a targeted change in the properties of the catalyst, including its specificity (especially in relation to macromolecular substrates), the dependence of catalytic activity on pH, ionic composition and other environmental parameters and, very importantly, its stability with respect to to various types of denaturing influences. Note that a major contribution to the development general principles stabilization of enzymes was done by Soviet researchers.

Fourthly, the immobilization of enzymes makes it possible to regulate their catalytic activity by changing the properties of the carrier under the influence of certain physical factors, such as light or sound. On this basis, mechanical and sound-sensitive sensors, amplifiers of weak signals and silver-free photographic processes are created.

As a result of the introduction of a new class of bioorganic catalysts - immobilized enzymes, new, previously inaccessible development paths have opened up for applied enzymology. Just listing the areas in which immobilized enzymes are used could take up a lot of space.

6. Directions for improving biological objects by methods of selection and mutagenesis. Mutagens. Classification. Characteristic. The mechanism of their action.

That mutations are the primary source of variability in organisms, creating the basis for evolution. However, in the second half of the 19th century. Another source of variability was discovered for microorganisms - the transfer of foreign genes - a kind of “genetic engineering of nature”.

For a long time, the concept of mutation was attributed only to chromosomes in prokaryotes and chromosomes (nucleus) in eukaryotes. Currently, in addition to chromosomal mutations, the concept of cytoplasmic mutations has also appeared (plasmid - in prokaryotes, mitochondrial and plasmid - in eukaryotes).

Mutations can be caused by both rearrangement of the replicon (change in the number and order of genes in it) and changes within an individual gene.

In relation to any biological objects, but especially often in the case of microorganisms, so-called spontaneous mutations are detected, which are found in a population of cells without special influence on it.

Based on the severity of almost any characteristic, cells in a microbial population form a variation series. Most cells have an average expression of the trait. Deviations “+” and “–” from the average value are less common in the population, the greater the deviation in any direction (Fig. I). The initial, simplest approach to improving a biological object was to select deviations “+” (assuming that these deviations correspond to the interests of production). In a new clone (genetically homogeneous offspring of one cell; on a solid medium - a colony), obtained from a cell with a “+” deviation, selection was again carried out according to the same principle. However, this procedure, when repeated several times, quickly loses its effectiveness, i.e., the “+” deviations become less and less in magnitude in new clones.

Mutagenesis is carried out when a biological object is treated with physical or chemical mutagens. In the first case, as a rule, these are ultraviolet, gamma, and x-rays; in the second - nitrosomethylurea, nitrosoguanidine, acridine dyes, some natural substances (for example, from DNA-tropic antibiotics due to their toxicity for infectious diseases not used in clinical practice). The mechanism of activity of both physical and chemical mutagens is associated with their direct effect on DNA (primarily on the nitrogenous bases of DNA, which is expressed in cross-linking, dimerization, alkylation of the latter, intercalation between them).

It is understood, of course, that the damage does not lead to death. Thus, after treating a biological object with mutagens (physical or chemical), their effect on DNA leads to frequent hereditary changes already at the level of the phenotype (certain of its properties). The next task is to select and evaluate the mutations that the biotechnologist needs. To identify them, the treated culture is sown on solid nutrient media of different compositions, having previously diluted it in such a way that there is no continuous growth on the solid medium, but separate colonies are formed, formed during the reproduction of individual cells. Then each colony is subcultured and the resulting culture (clone) is checked for certain characteristics in comparison with the original one. This selection part of the work as a whole is very labor-intensive, although techniques to increase its efficiency are constantly being improved.

Thus, by changing the composition of solid nutrient media on which colonies grow, one can immediately obtain initial information about the properties of the cells of this colony in comparison with the cells of the original culture. To seed clones with different metabolic characteristics, the so-called “fingerprint method”, developed by J. Lederberg and E. Lederberg, is used. The population of microbial cells is bred so that about a hundred colonies grow on a Petri dish with a nutrient medium and they are clearly separated. Velvet is placed on a metal cylinder with a diameter close to the diameter of the Petri dish; everything is then sterilized, thus creating a “sterile velvet bottom” of the cylinder. Next, apply this bottom to the surface of the medium in a cup with colonies grown on it. In this case, the colonies seem to “imprint” on the velvet. This velvet is then applied to the surface of media of different compositions. In this way, it is possible to establish: which of the colonies in the original dish (on velvet, the location of the colonies reflects their location on the surface of the solid medium in the original dish) corresponds, for example, to a mutant requiring a specific vitamin or a specific amino acid; or which colony consists of mutant cells capable of producing an enzyme that oxidizes a specific substrate; or which colony consists of cells that have become resistant to a particular antibiotic, etc.

First of all, the biotechnologist is interested in mutant crops that have an increased ability to form the target product. The producer of the target substance, the most promising in practical terms, can be repeatedly treated with different mutagens. New mutant strains obtained in scientific laboratories different countries world, serve as an object of exchange in creative cooperation, licensed sales, etc.

The potential of mutagenesis (with subsequent selection) is due to the dependence of the biosynthesis of the target product on many metabolic processes in the producer’s body. For example, increased activity of the organism that forms the target product can be expected if the mutation leads to duplication (doubling) or amplification (multiplication) of structural genes included in the system for synthesizing the target product. Further activity can be increased if, due to various types of mutations, the functions of repressor genes that regulate the synthesis of the target product are suppressed. A very effective way to increase the formation of the target product is to disrupt the retroinhibition system. It is also possible to increase the activity of a producer by changing (due to mutations) the system of transport of precursors of the target product into the cell. Finally, sometimes the target product, with a sharp increase in its formation, negatively affects the viability of its own producer (the so-called suicidal effect). Increasing the resistance of a producer to the substance it produces is often necessary to obtain, for example, superproducers of antibiotics.

In addition to duplication and amplification of structural genes, mutations can be of the nature of deletion - “erasure”, i.e. “loss” of part of the genetic material. Mutations can be caused by transposition (insertion of a section of a chromosome into a new location) or inversion (change in the order of genes on a chromosome). In this case, the genome of the mutant organism undergoes changes, leading in some cases to the loss of a certain trait by the mutant, and in others to the emergence of a new trait. Genes in new places are under the control of other regulatory systems. In addition, hybrid proteins unusual for the original organism may appear in mutant cells due to the fact that polynucleotide chains of two (or more) structural genes that were previously distant from one another are under the control of one promoter.

So-called “point” mutations can also be of considerable importance for biotechnological production. In this case, changes occur within only one gene. For example, the loss or insertion of one or more bases. “Point” mutations include transversion (when a purine is replaced by a pyrimidine) and transition (one purine is replaced by another purine or one pyrimidine by another pyrimidine). Substitutions in one pair of nucleotides (minimal substitutions) during the transmission of the genetic code at the translation stage lead to the appearance in the encoded protein of one amino acid of another. This can dramatically change the conformation of a given protein and, accordingly, its functional activity, especially in the case of replacement of an amino acid residue in the active or allosteric center.

One of the most brilliant examples of the effectiveness of mutagenesis followed by selection based on increasing the formation of the target product is the history of the creation of modern penicillin superproducers. Working with initial biological objects - strains (a strain is a clonal culture, the homogeneity of which certain signs supported by selection) of the fungus Penicillium chrysogenum isolated from natural sources, has been carried out since the 1940s. for several decades in many laboratories. Initially, some success was achieved in selecting mutants that resulted from spontaneous mutations. Then they moved on to inducing mutations with physical and chemical mutagens. As a result of a series of successful mutations and stepwise selection of increasingly productive mutants, the activity of Penicillium chrysogenum strains used in the industry of countries where penicillin is produced is now 100 thousand times higher than that of the original strain discovered by A. Fleming, from which the history of the discovery of penicillin began .

Industrial strains (in relation to biotechnological production) with such high productivity (this applies not only to penicillin, but also to other target products) are extremely unstable due to the fact that numerous artificial changes in the genome of the cells of the strain in themselves do not have a positive effect on the viability of these cells have. Therefore, mutant strains require constant monitoring during storage: the cell population is plated on a solid medium and cultures obtained from individual colonies are tested for productivity. In this case, revertants - cultures with reduced activity - are discarded. Reversion is explained by reverse spontaneous mutations leading to the return of a section of the genome (a specific fragment of DNA) to its original state. Special enzyme repair systems are involved in reversion to the norm - in the evolutionary mechanism of maintaining the constancy of the species.

Improving biological objects in relation to production is not limited to increasing their productivity. Although this direction is undoubtedly the main one, it cannot be the only one: the successful operation of biotechnological production is determined by many factors. From an economic point of view, it is very important to obtain mutants capable of using cheaper and less scarce nutrient media. If for work in a research laboratory expensive media do not create any special financial problems, then for large-scale production, reducing their cost (albeit without increasing the level of activity of the producer) is extremely important.

Another example: in the case of some biological objects, the culture liquid after the end of fermentation has technologically unfavorable rheological properties. Therefore, in the shop for isolating and purifying the target product, working with a culture liquid of high viscosity, they encounter difficulties when using separators, filter presses, etc. Mutations that appropriately change the metabolism of a biological object largely eliminate these difficulties.

The production of phage-resistant biological objects is of great importance in terms of guaranteeing the reliability of production. Compliance with aseptic conditions during fermentation primarily concerns the prevention of cells and spores of foreign bacteria and fibs (in more rare cases, algae and protozoa) from entering the seed material (as well as the fermentation apparatus). It is extremely difficult to prevent phages from entering the fermenter along with process air that is sterilized by filtration. It is no coincidence that viruses were called “filterable” in the first years after their discovery. Therefore, the main way to combat bacteriophages, actinophages and phages that infect fungi is to obtain mutant forms of biological objects that are resistant to them.

Without touching on special cases of working with biological objects-pathogens, it should be emphasized that sometimes the task of improving biological objects comes from the requirements of industrial hygiene. For example, a producer of one of the important betalactam antibiotics isolated from a natural source significant amount formed volatile substances with an unpleasant odor of rotting vegetables.

Mutations leading to the removal of genes encoding enzymes involved in the synthesis of these volatile substances have in this case acquired practical significance for production.

From all of the above it follows that a modern biological object used in the biotechnological industry is a super-producer, differing from the original natural strain not in one, but, as a rule, in several indicators. Storing such super-producing strains poses a serious independent problem. With all storage methods, they must be periodically reseeded and checked both for productivity and for other properties important for production.

In the case of using higher plants and animals as biological objects for the production of medicines, the possibilities of using mutagenesis and selection for their improvement are limited. However, in principle, mutagenesis and selection are not excluded here. This especially applies to plants that form secondary metabolites that are used as medicinal substances.

7. Directions for creating new biological objects using genetic engineering methods. Basic levels of genetic engineering. Characteristic.

Using genetic engineering methods, it is possible to construct according to a specific plan new forms of microorganisms capable of synthesizing a wide variety of products, including products of animal and plant origin. In this case, one should take into account the high growth rates and productivity of microorganisms, their ability to utilize various types of raw materials. The possibility of microbiological synthesis of human proteins opens up broad prospects for biotechnology: somatostatin, interferons, insulin, and growth hormone are obtained in this way.

The main problems on the way to designing new microorganisms-producers come down to the following.

1. Gene products of plant, animal and human origin enter an intracellular environment that is alien to them, where they are destroyed by microbial proteases. Short peptides such as somatostatin are hydrolyzed especially quickly, within a few minutes. The strategy for protecting genetically engineered proteins in a microbial cell comes down to: a) the use of protease inhibitors; Thus, the yield of human interferon increased 4 times when a DNA fragment of the T4 phage with the gene was introduced into the plasmid carrying the interferon gene pin, responsible for the synthesis of protease inhibitor; b) obtaining the peptide of interest as part of a hybrid protein molecule; for this purpose, the peptide gene is cross-linked with the natural gene of the recipient organism; the protein A gene is most often used Staphylococcus aureus\ c) amplification (increase in copy number) of genes; multiple repetitions of the human proinsulin gene as part of a plasmid led to synthesis in the cell E. coli multimer of this protein, which turned out to be much more stable to the action of intracellular proteases than monomeric proinsulin. The problem of stabilizing foreign proteins in cells has not yet been sufficiently studied (V.I. Tanyashin, 1985).

2. In most cases, the transplanted gene product is not released into the culture medium and accumulates inside the cell, which significantly complicates its isolation. Thus, the accepted method of producing insulin using E. coli involves cell destruction and subsequent purification of insulin. In this regard, great importance is attached to the transplantation of genes responsible for the excretion of proteins from cells. There is information about a new method of genetically engineered synthesis of insulin, which is released into the culture medium (M. Sun, 1983).

The reorientation of biotechnologists from their favorite object of genetic engineering is also justified E. coli to other biological objects. E. coli excretes relatively few proteins. In addition, the cell wall of this bacterium contains a toxic substance called endocotin, which must be carefully separated from products used for pharmacological purposes. Gram-positive bacteria (representatives of the genera Bacillus, Staphylococcus, Streptomyces). In particular Bas. subtilis releases more than 50 different proteins into the culture medium (S. Vard, 1984). These include enzymes, insecticides, and antibiotics. Eukaryotic organisms are also promising. They have a number of advantages, in particular, yeast interferon is synthesized in glycolyzed form, like native human protein (unlike interferon synthesized in cells E. coti).

3. Most hereditary traits are encoded by several genes, and genetic engineering development should include stages of sequential transplantation of each of the genes. An example of a multigene project implemented is the creation of a strain Pseudomonas sp., capable of utilizing crude oil. With the help of plasmids, the strain was successively enriched with genes for enzymes that break down octane, camphor, xylene, and naphthalene (V. G. Debabov, 1982). In some cases, not sequential, but simultaneous transplantation of entire blocks of genes using one plasmid is possible. As part of one plasmid, the nif operon can be transferred into the recipient cell Klebsiella pneumonia responsible for nitrogen fixation. The body's ability to fix nitrogen is determined by the presence of at least 17 different genes responsible for both the structural components of the nitrogenase complex and the regulation of their synthesis.

Genetic engineering of plants is carried out at the organismal, tissue and cellular levels. The demonstrated possibility, albeit for a few species (tomato, tobacco, alfalfa), of the regeneration of an entire organism from a single cell has sharply increased interest in plant genetic engineering. However, here, in addition to purely technical ones, it is necessary to solve problems associated with violations of the genome structure (changes in ploidy, chromosomal rearrangements) of cultivated plant cells. An example of a implemented genetic engineering project is the synthesis of phaseolin, a bean storage protein, in regenerated tobacco plants. Transplantation of the gene responsible for the synthesis of phaseolin was carried out using a Ti plasmid as a vector. Using the Ti plasmid, the gene for resistance to the antibiotic neomycin was also transplanted into tobacco plants, and using the CMV virus, the gene for resistance to the dihydrofolate reductase inhibitor methotrexate was transplanted into turnip plants.

Genetic engineering of plants includes manipulations not only with the nuclear genome of cells, but also with the genome of chloroplasts and mitochondria. It is in the chloroplast genome that it is most advisable to introduce a nitrogen fixation gene to eliminate the need of plants for nitrogen fertilizers. Two plasmids (S-1 and S-2) were found in maize mitochondria, causing cytoplasmic male sterility. If plant breeders need to “prohibit” corn from self-pollinating and allow only cross-pollination, they may not bother removing the stamens manually if they select cytoplasmically male-sterile plants for fertilization. Such plants can be developed through long-term selection, but genetic engineering offers a faster and more targeted method - the direct introduction of plasmids into the mitochondria of maize cells. Developments in the field of genetic engineering of plants should also include genetic modification of plant symbionts - nodule bacteria of the genus Rhizobium. It is supposed to introduce into the cells of these bacteria using plasmids hup(hydrogen uptake) - a gene that naturally exists only in some strains of R. japonicum And R. leguminosarum. Nir-gen determines the absorption and utilization of hydrogen gas released during the functioning of the nitrogen-fixing enzyme complex of nodule bacteria. Hydrogen recyclization avoids the loss of reducing equivalents during symbiotic nitrogen fixation in nodules leguminous plants and significantly increase the productivity of these plants.

The use of genetic engineering methods to improve breeds of farm animals remains a distant goal. We are talking about increasing the efficiency of feed use, increasing fertility, the yield of milk and eggs, the resistance of animals to diseases, accelerating their growth, and improving the quality of meat. However, the genetics of all these traits in farm animals has not yet been elucidated, which hinders attempts at genetic manipulation in this area.

8. Cellular engineering and its use in the creation of microorganisms and plant cells. Protoplast fusion method.

Cell engineering is one of the most important areas in biotechnology. It is based on the use of a fundamentally new object - an isolated culture of cells or tissues of eukaryotic organisms, as well as on totipotency - a unique property of plant cells. The use of this object has revealed great possibilities in solving global theoretical and practical problems. In the field of fundamental sciences, it has become feasible to study such complex problems as the interaction of cells in tissues, cell differentiation, morphogenesis, the implementation of cell totipotency, mechanisms of the appearance of cancer cells, etc. When solving practical problems, the main attention is paid to issues of selection, obtaining significant quantities of biologically valuable metabolites plant origin, in particular cheaper medicines, as well as the cultivation of healthy virus-free plants, their clonal propagation, etc.

In 1955, after the discovery by F. Skoog and S. Miller of a new class of phytohormones - cytokinins - it turned out that when they act together with another class of phytohormones - auxins - it became possible to stimulate cell division, maintain the growth of callus tissue, and induce morphogenesis under controlled conditions.

In 1959, a method for growing large masses of cell suspensions was proposed. An important event was the development by E. Cocking (University of Nottingham, UK) in 1960 of a method for obtaining isolated protoplasts. This served as an impetus for the production of somatic hybrids, the introduction of viral RNA, cellular organelles, and prokaryotic cells into protoplasts. At the same time, J. Morel and R. G. Butenko proposed a method of clonal micropropagation, which immediately found wide practical application. A very important advance in the development of technologies for culturing isolated tissues and cells has been the cultivation of a single cell using “nanny” tissue. This method was developed in Russia in 1969 at the Institute of Plant Physiology named after. K. A. Timiryazev RAS under the leadership of R. G. Butenko. In recent decades, rapid progress in cell engineering technologies has continued, making it possible to significantly facilitate breeding work. Great progress has been made in the development of methods for producing transgenic plants, technologies for using isolated tissues and cells of herbaceous plants, and the cultivation of tissues of woody plants has begun.

The term “isolated protoplasts” was first proposed by D. Hanstein in 1880. A protoplast in a whole cell can be observed during plasmolysis. An isolated protoplast is the contents of a plant cell surrounded by a plasmalemma. This formation does not have a cellulose wall. Isolated protoplasts are one of the most valuable objects in biotechnology. They make it possible to study various properties of membranes, as well as the transport of substances through the plasmalemma. Their main advantage is that it is quite easy to introduce genetic information from organelles and cells of other plants, prokaryotic organisms and animal cells into isolated protoplasts. E. Cocking established that an isolated protoplast, thanks to the mechanism of pinocytosis, is capable of absorbing from the environment not only low molecular weight substances, but also large molecules, particles (viruses) and even isolated organelles.

Of great importance in the creation of new plant forms for studying the interaction of the nuclear genome and organelle genomes is the ability of isolated protoplasts to merge, forming hybrid cells. In this way, it is possible to obtain hybrids from plants with varying degrees of taxonomic distance, but with valuable economic qualities.

Protoplasts were first isolated by J. Klerner in 1892 while studying plasmolysis in telores leaf cells (Stratiotes aloides) during mechanical damage fabrics. Therefore, this method is called mechanical. It allows you to highlight only a large number of protoplasts (production is not possible from all types of tissues); the method itself is long and labor-intensive. The modern method of isolating protoplasts involves removing the cell wall using the step-by-step use of enzymes to destroy it: cellulases, hemicellulases, pectinase. This method is called enzymatic.

The first successful isolation of protoplasts from higher plant cells using this method was made by E. Cocking in 1960. Compared to the mechanical enzymatic method, it has a number of advantages. It makes it possible to isolate a large number of protoplasts relatively easily and quickly, and they do not experience strong osmotic shock. After the action of the enzymes, the mixture of protoplasts is passed through a filter and centrifuged to remove undestroyed cells and their fragments.

Protoplasts can be isolated from plant tissue cells, callus cultures and suspension cultures. Optimal conditions for the isolation of protoplasts are individual for different objects, which requires painstaking preliminary work on the selection of enzyme concentrations, their ratio, and processing time. A very important factor allowing the isolation of whole viable protoplasts is the selection of an osmotic stabilizer. Various sugars are usually used as stabilizers, sometimes ionic osmotics (solutions of salts CaCl 2, Na 2 HP0 4, KCI). The osmotic concentration must be slightly hypertonic so that the protoplasts are in a state of weak plasmolysis. In this case, metabolism and cell wall regeneration are inhibited.

Isolated protoplasts can be cultured. Typically, the same media on which isolated cells and tissues grow are used for this purpose. Immediately after the removal of enzymes from protoplasts in culture, the formation of a cell wall begins. The protoplast that has regenerated the wall behaves like an isolated cell and is capable of dividing and forming a clone of cells. Regeneration of whole plants from isolated protoplasts is associated with a number of difficulties. So far, it has been possible to obtain regeneration through embryogenesis only in carrot plants. By stimulating the sequential formation of roots and shoots (organogenesis), the regeneration of tobacco, petunia and some other plants was achieved. It should be noted that protoplasts isolated from a genetically stable cell culture more often regenerate plants and are used with great success in studies of genetic modification of protoplasts.

9. Methods of cell engineering as applied to animal cells. Hybridoma technology and its use in biotechnological processes.

In 1975, G. Köhler and K. Milstein were able for the first time to isolate cell clones capable of secreting only one type of antibody molecules and at the same time growing in culture. These cell clones were obtained by the fusion of antibody-forming and tumor cells - chimera cells, called hybridomas, since, on the one hand, they inherited the ability for almost unlimited growth in culture, and on the other hand, the ability to produce antibodies of a certain specificity (monoclonal antibodies) .

It is very important for a biotechnologist that selected clones can be stored frozen for a long time, so if necessary, a certain dose of such a clone can be taken and administered to an animal that will develop a tumor producing monoclonal antibodies of a given specificity. Soon antibodies will be detected in the animal's serum in a very high concentration of 10 to 30 mg/ml. Cells of such a clone can also be grown in vitro, and the antibodies they secrete can be obtained from the culture liquid.

The creation of hybridomas that can be stored frozen (cryopreservation) made it possible to organize entire hybridoma banks, which in turn opened up great prospects for the use of monoclonal antibodies. Their scope of application, in addition to the quantitative determination of various substances, includes a wide variety of diagnostics, for example, the identification of a specific hormone, viral or bacterial antigens, blood group antigens and tissue antigens.

Stages of obtaining hybrid cells. Cell fusion is preceded by the establishment of close contact between the plasma membranes. This is prevented by the presence of a surface charge on natural membranes, caused by negatively charged groups of proteins and lipids. Depolarization of membranes by an alternating electric or magnetic field, neutralization of the negative charge of membranes with the help of cations promotes cell fusion. In practice, Ca2+ ions and chlorpromazine are widely used. An effective “merging” (fusogenic) agent is polyethylene glycol.

The Sendai virus is also used in relation to animal cells, the action of which as a fusion agent is apparently associated with partial hydrolysis of proteins of the cytoplasmic membrane. The FI subunit region of the virus has proteolytic activity (S. Nicolau et al., 1984). Before fusion, plant, fungal and bacterial cells are freed from the cell wall, resulting in protoplasts. The cell wall is subjected to enzymatic hydrolysis using lysozyme (for bacterial cells), snail zymolyase (for fungal cells), a complex of cellulases, hemicellulases and pectinases produced by fungi (for plant cells). Swelling and subsequent destruction of protoplasts is prevented by creating an increased osmolarity of the medium. The selection of hydrolytic enzymes and salt concentrations in the medium in order to ensure maximum yield of protoplasts is difficult task, which is decided in each case separately.

To screen the resulting hybrid cells, various approaches are used: 1) taking into account phenotypic characteristics; 2) creation of selective conditions in which only hybrids that combine the genomes of parental cells survive.

Possibilities of the cell fusion method. The method of somatic cell fusion opens up significant prospects for biotechnology.

1. The possibility of crossing phylogenetically distant forms of living things. By fusion of plant cells, fertile, phenotypically normal interspecific hybrids of tobacco, potatoes, cabbage with turnips (equivalent to natural rapeseed), and petunias were obtained. There are sterile intertribal hybrids of potato and tomato, sterile intertribal hybrids of Arabidopsis and turnip, tobacco and potato, tobacco and belladonna, which form morphologically abnormal stems and plants. Cell hybrids have been obtained between representatives of different families, existing, however, only as disorganized growing cells (tobacco and peas, tobacco and soybeans, tobacco and faba beans). Interspecific (Saccharomyces uvarum and S. diastalicus) and intergeneric (Kluyveromyces lactis and S. cerevisiae) yeast hybrids were obtained. There is evidence of the fusion of cells of various types of fungi and bacteria.

Experiments on the fusion of cells of organisms belonging to different kingdoms, for example, cells of the Xenopus taevis frog and carrot protoplasts, seem somewhat curious. A hybrid plant-animal cell gradually becomes encased in a cell wall and grows on media on which plant cells are cultured. The nucleus of an animal cell, apparently, quickly loses its activity (E. S. Cocking, 1984).

2. Obtaining asymmetric hybrids carrying the full set of genes of one of the parents and a partial set of the other parent. Such hybrids often arise from the fusion of cells of organisms that are phylogenetically distant from each other. In this case, due to incorrect cell divisions caused by the uncoordinated behavior of two dissimilar sets of chromosomes, the chromosomes of one of the parents are partially or completely lost in a series of generations.

Asymmetrical hybrids are more stable, more fertile and more viable than symmetrical ones, which carry complete sets of genes from the parent cells. For the purpose of asymmetric hybridization, it is possible to selectively treat the cells of one of the parents to destroy part of its chromosomes. Targeted transfer from cell to cell of the desired chromosome is possible. It is also of interest to obtain cells in which only the cytoplasm is hybrid. Cytoplasmic hybrids are formed when, after cell fusion, the nuclei retain their autonomy and, upon subsequent division of the hybrid cell, end up in different daughter cells. Screening of such cells is carried out using marker genes of the nuclear and cytoplasmic (mitochondrial and chloroplast) genomes.

Cells with fused cytoplasm (but not nuclei) contain the nuclear genome of one of the parents and at the same time combine the cytoplasmic genes of the fused cells. There are indications of DNA recombination of mitochondria and chloroplasts in hybrid cells.

Obtaining hybrids by merging three or more parent cells. Regenerated plants (fungi) can be grown from such hybrid cells.

Hybridization of cells carrying different developmental programs is the fusion of cells of different tissues or organs, the fusion of normal cells with cells whose developmental program is changed as a result of malignant degeneration. In this case, so-called hybridoma cells are obtained, or hybridomas, which inherit from a normal parental cell the ability to synthesize one or another useful compound, and from a malignant one - the ability to grow rapidly and unlimitedly.

Hybridoma technology. The production of hybridomas today is the most promising direction cell engineering. The main goal is to “immortality” a cell that produces valuable substances by merging with a cancer cell and cloning the resulting hybridoma cell line. Hybridomas are obtained on the basis of cells - representatives of various kingdoms of life. The fusion of plant cells, which usually grow slowly in culture, with plant tumor cells makes it possible to obtain clones of fast-growing cells that produce the desired compounds. There are many applications of hybridoma technology to animal cells, where with its help it is planned to obtain unlimitedly multiplying producers of hormones and protein factors in the blood. Of greatest practical importance are hybridomas - products of the fusion of cells of malignant tumors of the immune system (myelomas) with normal cells of the same system - lymphocytes.

When a foreign agent enters the body of an animal or human - bacteria, viruses, “foreign” cells or simply complex organic compounds - lymphocytes are mobilized to neutralize the introduced agent. There are several populations of lymphocytes whose functions differ. There are so-called T-lymphocytes, among which are T-killers (“killers”), which directly attack a foreign agent in order to inactivate it, and B-lymphocytes, the main function of which is to produce immune proteins (immunoglobulins) that neutralize the foreign agent by binding with its surface areas (antigenic determinants), in other words, B lymphocytes produce immune proteins, which are antibodies to a foreign agent - antigen.

The fusion of a killer T-lymphocyte with a tumor cell produces a clone of unlimitedly multiplying cells that hunt for a specific antigen - the one to which the taken T-lymphocyte was specific. They are trying to use similar T-killer hybridoma clones to fight cancer cells directly in the patient’s body (B. Fuchs et al., 1981; 1983),

When a B-lymphocyte fuses with a myeloma cell, B-hybridoma clones are obtained, which are widely used as producers of antibodies targeting the same antigen as the antibodies synthesized by the B-lymphocyte that gave rise to the clone, i.e., monoclonal antibodies. Monoclonal antibodies are homogeneous in their properties; they have the same affinity for the antigen and bind to. one single antigenic determinant. This is an important advantage of monoclonal antibodies - products of B-hybridome, compared to antibodies obtained without the use of cell engineering, by immunizing a laboratory animal with a selected antigen with subsequent isolation of antibodies from its blood serum or as a result of direct interaction of the antigen with a population of lymphocytes in tissue culture . Similar traditional methods produce a mixture of antibodies that differ in specificity and affinity for the antigen, which is explained by the participation in the production of antibodies of many different clones of B-lymphocytes and the presence of several determinants in the antigen, each of which corresponds to a special type of antibody. Thus, monoclonal antibodies selectively bind to only one antigen, inactivating it, which is of great practical importance for the recognition and treatment of diseases caused by foreign agents - bacteria, fungi, viruses, toxins, allergens and transformed self cells (cancer tumors). Monoclonal antibodies successfully used for analytical purposes to study cellular organelles, their structure or individual biomolecules.

Until recently, only mouse and rat myeloma cells and B lymphocytes were used for hybridization. The monoclonal antibodies they produce have limited therapeutic use, since they themselves are a foreign protein for the human body. Mastering the technology for producing hybridomas based on human immune cells is associated with significant difficulties: human hybridomas grow slowly and are relatively little stable. However, human hybridomas have already been obtained - producers of monoclonal antibodies. It turned out that human monoclonal antibodies in some cases cause immune reactions, and their clinical effectiveness depends on the correct selection of the class of antibodies and hybridoma lines suitable for a given patient. The advantages of human monoclonal antibodies include the ability to recognize subtle differences in antigen structure that are not recognized by mouse or rat monoclonal antibodies. Attempts have been made to obtain chimeric hybridomas combining mouse myeloma cells and human B lymphocytes; such hybridomas have so far found only limited use (tK-Haron, 1984).

Along with undoubted advantages, monoclonal antibodies also have disadvantages that create problems in their practical use. They are not stable when stored in a dried state, while at the same time, a mixture of conventional (polyclonal) antibodies always contains a group of antibodies that are stable under selected storage conditions. Thus, the heterogeneity of conventional antibodies gives them an additional reserve of stability when external conditions change, which corresponds to one of the basic principles of increasing the reliability of systems. Monoclonal antibodies often have too low an affinity for an antigen and an overly narrow specificity, which prevents their use against the variable antigens characteristic of infectious agents and tumor cells. It should also be noted that high cost monoclonal antibodies on the international market.

General scheme obtaining hybridomas based on myeloma cells and immune lymphocytes includes the following steps.

1. Obtaining mutant tumor cells that die during subsequent selection of hybridoma cells. The standard approach is to develop myeloma cell lines that are incapable of synthesizing enzymes in the spare pathways for the biosynthesis of purines and pyrimidines from hypoxanthine and thymidine, respectively (Figure 6). The selection of such tumor cell mutants is carried out using toxic analogues of hypoxanthine and thymidine. In a medium containing these analogs, only mutant cells that lack the enzymes hypoxanthine guanine phosphoribosyltransferase and thymidine kinase, which are necessary for the reserve pathways of nucleotide biosynthesis, survive.

Do you have any idea what biotechnology is?

Of course, you have heard something about them. This is an innovative direction in modern biology, which is on a par with such sciences as mathematics or physics.

Biotechnology deals with the creation of products and materials that people need using living cultures and microorganisms such as yeast, fungal spores, cultivated cells of plants and animals, etc. The construction of the necessary genes using genetic and cellular engineering methods makes it possible to control the heredity and vital activity of animals, plants and microorganisms and create organisms with new properties beneficial to humans that have not previously been observed in nature. Bioengineers deal with living systems of nature, using their capabilities to solve medical problems, genetic engineering, agriculture, the chemical industry, the cosmetics industry and the food industry. Biotechnology is a science at the intersection of related industries.

It is interesting that the penetration of biotechnology into the world economy is reflected in the fact that new terms have been formed to denote the global nature of this process. Multi-colored biotechnologies have even appeared in industry:

• “red” biotechnology – biotechnology associated with ensuring human health and potential correction of its genome, as well as with the production of biopharmaceuticals (proteins, enzymes, antibodies);
• "green" biotechnology - aimed at the development and creation of genetically modified (GM) plants that are resistant to biotic and abiotic stresses, determines modern methods of agriculture and forestry;
• “white” - industrial biotechnology, combining the production of biofuel, biotechnology in the food, chemical and oil refining industries;
• “gray” - associated with environmental protection activities, bioremediation;
• “blue” biotechnology is associated with the use of marine organisms and raw materials.

New professions have also appeared: biopharmacologist, bionicist, architect of living systems, urban ecologist and others. Well, the economy that unites all these innovative areas began to be called “bioeconomics”.

Today, our country lags behind the countries that are technological leaders in this area in terms of production based on high biotechnologies. Our state’s policy on import substitution is aimed precisely at not only creating new biotechnologies, but also transferring foreign solutions that have already received recognition in the world to our country.

Technology transfer is accompanied by the search for the newest and most progressive solutions. But there is one important point, in addition to the fact that technology is progressive today, you need to be able to predict its prospects for the progress of the future.

Sometimes entire research institutes, groups of scientists and practitioners work to make such strategic predictions. And sometimes, only one person can predict the prospects and breakthrough nature of a technology. Like Steve Jobs or Bill Geitz.

The biotech industry also has its share of insightful business leaders. One of them is Yakovlev Maxim Nikolaevich, general director of the representative office of the biotechnological corporation Unhwa, South Korea, located in the city of St. Petersburg.

Biotechnology, which Maxim Yakovlev has identified as a breakthrough future in various segments of the economy, is in the field of cultivating plant cells that have the functions of “natural biofactories” for the production of valuable ingredients from any plant, including unique ones.

This promising biotechnology, according to the businessman, is capable of creating natural food from one isolated plant cell directly on board spaceships, growing fruits and vegetables with the desired characteristics and sizes, creating ecosystems of other planets and food for humans on an industrial scale from any plant without cultivation of this plant on living soil.

Perhaps such prospects for biotechnology are still difficult to comprehend and accept as possible. But we all know that there are people who are able to see beyond the masses, because they themselves already live in the future and call us to follow them.

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The term “biotechnology” was first used by the Hungarian engineer Karl Ereky in 1917. Certain elements of biotechnology appeared quite a long time ago. In essence, these were attempts to use individual cells (microorganisms) and some enzymes in industrial production to facilitate the occurrence of a number of chemical processes.

Thus, in 1814, St. Petersburg academician K. S. Kirchhoff discovered the phenomenon of biological catalysis and tried to obtain sugar from available domestic raw materials using a biocatalytic method (until the mid-19th century, sugar was obtained only from sugar cane). In 1891, in the USA, the Japanese biochemist Dz. Takamine received the first patent for the use of enzyme preparations for industrial purposes: the scientist proposed using diastase for the saccharification of plant waste.

At the beginning of the 20th century, the fermentation and microbiological industries actively developed. During these same years, the first attempts were made to use enzymes in the textile industry.

In 1916–1917, the Russian biochemist A. M. Kolenev tried to develop a method that would make it possible to control the action of enzymes in natural raw materials during the production of tobacco.

A huge contribution to the practical use of biochemistry achievements was made by Academician A. N. Bakh, who created an important applied area of ​​biochemistry - technical biochemistry. A. N. Bach and his students developed many recommendations for improving technologies for processing a wide variety of biochemical raw materials, improving technologies for baking, brewing, winemaking, tea and tobacco production, etc., as well as recommendations for increasing the yield of cultivated plants by managing them by biochemical processes.

All these studies, as well as the progress of the chemical and microbiological industries and the creation of new industrial biochemical production (tea, tobacco, etc.) were the most important prerequisites for the emergence of modern biotechnology.

In production terms, the microbiological industry became the basis of biotechnology in the process of its formation. Behind post-war years The microbiological industry acquired fundamentally new features: microorganisms began to be used not only as a means of increasing the intensity of biochemical processes, but also as miniature synthetic factories capable of synthesizing the most valuable and complex chemical compounds inside their cells. The turning point was associated with the discovery and start of production of antibiotics.

The first antibiotic, penicillin, was isolated in 1940. Following penicillin, other antibiotics were discovered (this work continues to this day). With the discovery of antibiotics, new tasks immediately appeared: establishing the production of medicinal substances produced by microorganisms, working to reduce the cost and increase the availability of new drugs, and obtaining them in very large quantities needed by medicine.

Synthesizing antibiotics chemically was very expensive or even incredibly difficult, almost impossible (it is not without reason that the chemical synthesis of tetracycline by the Soviet scientist Academician M. M. Shemyakin is considered one of the largest achievements of organic synthesis). And then they decided to use microorganisms that synthesize penicillin and other antibiotics for the industrial production of drugs. This is how the most important area of ​​biotechnology arose, based on the use of microbiological synthesis processes.

Types of biotechnology

Bioengineering

Bioengineering or biomedical engineering is a discipline aimed at advancing the knowledge of engineering, biology and medicine and improving human health through interdisciplinary developments that combine engineering approaches with advances in biomedical science and clinical practice. Bioengineering/biomedical engineering is the application of engineering approaches to solve medical problems to improve health care. This engineering discipline focuses on using knowledge and experience to find and solve problems in biology and medicine.

Bioengineers work for the benefit of humanity, dealing with living systems and applying advanced technologies to solve medical problems. Biomedical engineering specialists can participate in the creation of devices and equipment, in the development of new procedures based on interdisciplinary knowledge, and in research aimed at obtaining new information to solve new problems.

Among the important achievements of bioengineering are the development of artificial joints, magnetic resonance imaging, pacemakers, arthroscopy, angioplasty, bioengineered skin prostheses, renal dialysis, and heart-lung machines. Also, one of the main areas of bioengineering research is the use of computer modeling methods to create proteins with new properties, as well as modeling the interaction of various compounds with cellular receptors in order to develop new pharmaceuticals (“drug design”).

Biomedicine

A branch of medicine that studies from a theoretical perspective the human body, its structure and function in normal and pathological conditions, pathological conditions, methods of their diagnosis, correction and treatment. Biomedicine includes accumulated information and research, to a greater or lesser extent, general medicine, veterinary medicine, dentistry and fundamental biological sciences, such as chemistry, biological chemistry, biology, histology, genetics, embryology, anatomy, physiology, pathology, biomedical engineering, zoology, botany and microbiology.

Monitoring, correcting, engineering and controlling human biological systems at the molecular level using nanodevices and nanostructures. A number of technologies for the nanomedicine industry have already been created in the world. These include targeted delivery of drugs to diseased cells, laboratories on a chip, and new bactericidal agents.

Biopharmacology

A branch of pharmacology that studies the physiological effects produced by substances of biological and biotechnological origin. In fact, biopharmacology is the fruit of the convergence of two traditional sciences - biotechnology, namely, that branch of it that is called “red”, medical biotechnology, and pharmacology, which was previously only interested in small-molecule chemicals, as a result of mutual interest.

Objects of biopharmacological research are the study of biopharmaceuticals, planning their production, organizing production. Biopharmacological therapeutic agents and means for the prevention of diseases are obtained using living biological systems, tissues of organisms and their derivatives, using biotechnology, that is, medicinal substances of biological and biotechnological origin.

Bioinformatics

A set of methods and approaches, including:

1. mathematical methods of computer analysis in comparative genomics (genomic bioinformatics);
2. development of algorithms and programs for predicting the spatial structure of proteins (structural bioinformatics);
3. research strategies, appropriate computational methodologies, and general management information complexity of biological systems.

Bioinformatics uses methods of applied mathematics, statistics and computer science. Bioinformatics is used in biochemistry, biophysics, ecology and other fields.

Bionics

Applied science about the application in technical devices and systems of the principles of organization, properties, functions and structures of living nature, that is, the forms of living things in nature and their industrial analogues. Simply put, bionics is a combination of biology and technology. Bionics looks at biology and technology from a completely new perspective, explaining what common features and what differences exist in nature and technology.

Distinguish:

• biological bionics, which studies the processes occurring in biological systems;
• theoretical bionics, which builds mathematical models of these processes;
• technical bionics, which applies models of theoretical bionics to solve engineering problems.

Bionics is closely related to biology, physics, chemistry, cybernetics and engineering sciences: electronics, navigation, communications, maritime science and others.

Bioremediation

A set of methods for purifying water, soil and atmosphere using the metabolic potential of biological objects - plants, fungi, insects, worms and other organisms.

Cloning

The appearance naturally or the production of several genetically identical organisms through asexual (including vegetative) reproduction. The term “cloning” in the same sense is often used in relation to the cells of multicellular organisms. Cloning is also called obtaining several identical copies of hereditary molecules (molecular cloning). Finally, cloning is also often referred to as biotechnological methods used to artificial production clones of organisms, cells or molecules. A group of genetically identical organisms or cells is a clone.

Genetic engineering

The essence of genetic engineering is the artificial creation of genes with the desired properties and their introduction into the appropriate cell. Gene transfer is carried out by a vector (recombinant DNA) - a special DNA molecule constructed from the DNA of viruses or plasmids, which contains the desired gene, transports it into the cell and ensures its integration into the genetic apparatus of the cell.

To mark certain cells of organisms in molecular genetic studies, the GFP gene isolated from jellyfish is used. It provides the synthesis of fluorescent protein, which glows in the dark.

Genetic engineering is widely used in both scientific research and the latest methods selection.

Biotechnology is a set of industrial methods that are used to produce various substances using living organisms, biological processes or phenomena. Traditional biotechnology is based on the phenomenon of fermentation - the use of microbial enzymes in production processes. Cellular engineering is a branch of biotechnology that develops and uses technologies for cultivating cells and tissues outside the body under artificial conditions. Genetic engineering is a branch of biotechnology that develops and uses technologies for isolating genes from organisms and individual cells, modifying them, and introducing them into other cells or organisms.

Some ethical and legal aspects of the use of biotechnological methods

Ethics is the doctrine of morality, according to which the main virtue is the ability to find a middle ground between two extremes. This science was founded by Aristotle.

Bioethics is a part of ethics that studies the moral side of human activity in medicine and biology. The term was proposed by V.R. Potter in 1969

In a narrow sense, bioethics refers to a range of ethical problems in the field of medicine. In a broad sense, bioethics refers to the study of social, environmental, medical and socio-legal problems affecting not only humans, but also any living organisms included in ecosystems. That is, it has a philosophical orientation, evaluates the results of the development of new technologies and ideas in medicine, biotechnology and biology in general.

Modern biotechnological methods have such powerful and not fully explored potential that their widespread use is possible only with strict adherence to ethical standards. The moral principles existing in society oblige us to seek a compromise between the interests of society and the individual. Moreover, the interests of the individual are currently being placed above the interests of society. Therefore, compliance with and further development of ethical standards in this area should be aimed, first of all, at the full protection of human interests.

The massive introduction into medical practice and commercialization of fundamentally new technologies in the field of genetic engineering and cloning has also led to the need to create an appropriate legal framework regulating all legal aspects of activities in these areas.

Let us dwell on those areas of biotechnological research that are directly related to a high risk of violation of individual rights and cause the most heated debate about their widespread use: transplantation of organs and cells for therapeutic purposes and cloning.

In recent years, there has been a sharp increase in interest in the study and use of human embryonic stem cells in biomedicine and cloning techniques to obtain them. As is known, embryonic stem cells are capable of transforming into different types of cells and tissues (hematopoietic, reproductive, muscle, nervous, etc.). They turned out to be promising for use in gene therapy, transplantology, hematology, veterinary medicine, pharmacotoxicology, drug testing, etc.

These cells are isolated from human embryos and fetuses of 5-8 weeks of development obtained during medical termination of pregnancy (as a result of abortion), which raises numerous questions regarding the ethical and legal legality of conducting research on human embryos, including the following:

• How necessary and justified is scientific research on human embryonic stem cells?
• Is it permissible to destroy human life for the sake of medical progress and how moral is this?
• Is the legal framework sufficiently developed for the use of these technologies?

In a number of countries, any research on embryos is prohibited (for example, in Austria, Germany). In France, the rights of the embryo are protected from the moment of conception. In the UK, Canada and Australia, although the creation of embryos for research purposes is not prohibited, a system of legislation has been developed to regulate and control such research.

In Russia, the situation in this area is more than uncertain: activities on the study and use of stem cells are not sufficiently regulated, and significant gaps remain in the legislation that hinder the development of this area. Regarding cloning, in 2002, federal law introduced a temporary (5-year) ban on human cloning, but it expired in 2007, and the issue remains open.

Biotechnology market

IT has many more parallels with modern biotech than it might seem at first glance. Information technologies did not appear on their own; their blossoming was preceded by fundamental discoveries in physics, physics of materials, computational mathematics and cybernetics. As a result, today IT is the area of ​​“easy startups”, in which very little time passes from the emergence of an idea to making a profit, and few people think about the work that has been done to date.

The situation with biotechnologies is similar, we are just now at an earlier stage, when tools and programs are still being developed. Biotechnologies are waiting for the appearance of their “personal computer,” only in our case it will not be an understandable mass device - we are talking more about a set of effective and inexpensive tools.

We can say that the situation now is similar to what it was in the 1990s in IT. Technologies are still developing and are quite expensive. For example, complete sequencing of a person costs $1000. This is much cheaper than the $3.3 billion price tag of the Human Genome Project, but it is still incredibly high for the average person, and its application for clinical diagnostics on a wide scale is not yet possible. To do this, the technology needs to fall in price by another factor of 10 and improve the technical properties so much that sequencing errors are leveled out. There are no such powerful projects in biotech as Facebook, but Illumina, Oxford Nanopore, Roche are all extremely successful companies, whose activities often resemble Google, which buys up interesting startups. Nanopore, for example, became billionaires before they even entered the market thanks to a combination of a good initial idea, management and success in raising funding.

Today, biotechnology is also a big data market, and this continues the parallels with IT, which in this case serves as a kind of tool for larger and more complex biotech. Companies such as Editas Medicine (one of the creators of the acclaimed CRISPR/Cas9 genome editing technology) have made their IP based on the results of sequencing bacterial genomic data from open sources. They were far from the first to reap the benefits of the accumulated information, they were not even the first to discover the operating principle of the CRISPR cluster, but it was Editas Medicine that created the biotechnological product. Today it is a company worth more than $1 billion.

And this is not the only business that will arise from the analysis of existing data. Moreover, it cannot be said that there is a queue for such data - there is already much more of it than can be analyzed, and there will be even more, because scientists do not stop sequencing. Unfortunately, analysis methods are still imperfect, so not everyone is able to turn data into a multi-billion dollar product. But if we estimate the rate of development of analysis tools (hint: it is very fast), it is not difficult to understand that in the future there will be many more companies that notice something interesting in big genomic data.

Can Russia become a biotech country?

The main problem of biotechnology in Russia is not the ban on GMOs, as many people think, but a large number of various bureaucratic barriers. This fact is also noted in the government. But even barriers can be adapted to. Over the past 26 years, we have been developing under the pressure of reforms, constant changes in the rules of the game, and business needs stability and confidence that no shocks will occur.

If Russian biotechnologies are not interfered with, they will begin to develop. I would also like to note that a thoughtless desire to help, those very ill-considered state investments, in fact, lead to the opposite result - subsidies teach companies that they will be constantly supported by the state. As practice shows, companies with state investments become ineffective. Healthy competition is needed everywhere, so initial contributions should not even come from the state, but from business, which should feel confident in the future, something we still have problems with.

The most correct thing for the state is to invest in creating an optimal environment for biotech. We have both minds and people with the energy and desire to create - it is important not to let this desire go to waste.

Today, biotechnologies are in a phase of intensive growth, but one can already imagine the vector of their development. After all, the very meaning of technology will not change, just as it did not change after the advent of the computer: its idea in 1951 was not very different from the one behind modern computers. Only functionality and performance differ significantly. The same thing will happen with biotechnologies, and the driver of their development is even clearer - this is the eternal desire of people to be healthy and live long, without observing all the complex rules healthy image life. Therefore, in the very near future we will see the rise of biotechnology, and ultimately this is great news for all of humanity.

Biotechnology (Βιοτεχνολογία, from Greek Bios- life, techne- art, craftsmanship and logos- word, doctrine) - the use of living organisms and biological processes in production. Biotechnology is an interdisciplinary field that arose at the intersection of biological, chemical and technical sciences. The development of biotechnology is associated with solving global problems of humanity - eliminating shortages of food, energy, mineral resources, improving the state of health care and the quality of the environment.

Method

A positive factor in the use of the biological method is its environmental friendliness. Biological agents can be used without limiting the frequency of application, while the number of treatments of plants with chemical pesticides is strictly regulated.

Biological plant protection is based on a systematic approach and integrated implementation of two main directions: conservation and promotion of the activities of natural populations of beneficial species (entomophages, microorganisms), self-defense of cultivated plants in agrobiocenoses and renewal of agrobiocenoses with useful species that are in short supply or absent. The fundamental difference between the biological method of plant protection and any other is the use of the first direction, which is carried out using biological preparations using methods of seasonal colonization, introduction and acclimatization of zoophages and microorganisms. The reproduction and efficiency of beneficial species is facilitated by agrobiotechnical measures, and some methods of soil cultivation with which you can create favorable conditions for the life activity of zoophages.

Cultivation of pest-resistant varieties of cultivated plants contributes to the formation of low-density pest populations.

Each of the main means of the biological method (the use of zoophages, useful in protecting plants by microorganisms) has its own characteristics and is effective in appropriate conditions. These means do not exclude, but complement each other. Now Special attention is focused on finding ways to combine biological protection with other methods in integrated plant protection systems from pests. The main objective of this method is to study the conditions that determine the effectiveness of natural enemies of pests and to develop ways to regulate their numbers and relationships with pest populations.

Introduction and acclimatization are used against quarantine pests that have limited distribution in the country.

Natural enemies limit the reproduction of the pest in its homeland, and they are absent in the new geographical area. Effective zoophages and microorganisms for importation and acclimatization are found in the homeland of the pest and relocated to new areas. Best results obtained by importing highly specialized species that are adapted to subsist on a single pest, disease, or weed. Intra-areal relocation involves the relocation of effective, often specialized, natural enemies from old foci, where the number of pests is declining, to new ones in other parts of the species’ range, where these enemies are absent or have not yet accumulated.

Microorganisms that damage harmful species are used to protect plants in the form of biological preparations. Most biological bacterial preparations are based on crystal-soluble bacteria of the Bacillus thuringiensis Berl. group, which form spores and crystals that can dissolve in the intestines of insects, where they enter with food.

Mushroom preparations contain spores of entomopathogenic fungi belonging to the imperfect ones.

Viral biological preparations (Verinykh) are made on the basis of polyhedrosis and granulosa viruses, which most often infect Lepidoptera.

In living systems at all levels of organization, a common method of transmitting information is chemical communication. Recently, much attention has been paid to the development and use of biologically active substances that ensure the relationship between living organisms in biocenoses, their growth and development. The main group of biologically active substances is pheromones. Pheromones are chemicals that insects produce and release into the environment. These substances cause corresponding behavioral or physiological reactions. There are different groups of pheromones - sex, aggregation, trace, etc. Sex pheromones, which are most often released by females to attract males, have become most widespread in plant protection practice. The most studied are the pheromones of lepidoptera, coleoptera, bedbugs, lacewings, and termites. Based on the determination of the structure of natural insect pheromones, their synthetic analogues have been created. Sex pheromones are used to detect and determine the distribution zone of pests, to signal the timing of the application of protective measures, determine the density of pest populations, as well as to protect crops by mass capture of males (“male vacuum”) and disorientation, attracting males during chemical sterilization.

The method of disorienting insects involves saturating an area with high concentrations of synthetic pheromone and disrupting pheromone communication between males and females. As a result, unmated females lay unfertilized eggs, which leads to a decrease in the population of the species. It has been established that the processes of metamorphosis, molting, reproduction and diapause of insects are regulated by hormones. The most studied are juvenile (larval), ecdysone (larval) and brain. Hormones were synthesized and obtained as chemical compounds, their structure differs from natural ones, but they imitate their biological activity - they act as regulators of the growth and development of insects. Chitin and juvenoid synthesis inhibitors have gained practical application in plant protection. Hormonal drugs their action differs significantly from traditional insecticides. They are not toxic, but cause disturbances in embryonic development, metamorphosis, and cause sterilization. Chitin inhibitors disrupt cuticle formation during molting. Juvenoids cause death upon completion of larval or doll development, and are inhibitors of chitin synthesis during the next Linqi.

The genetic method of combating pests was developed and proposed by A. S. Serebrovsky (1938, 1950). This method involves saturating the natural pest population with genetically inferior individuals of the same species. Females of a natural population, mating with such individuals, lay non-viable eggs, do not produce offspring, and the pest self-destructs. The genetic method is carried out by radiation and chemical sterilization. Radiation sterilization involves mass breeding of pests, irradiation of them (with gamma rays, X-rays) and subsequent release into fruit trees and crops. Damage to the chromosomal apparatus occurs in irradiated individuals. In chemical sterilization, sterilizers use chemicals with alkyl compounds, antimetabolites and antibiotics. The former cause sexual sterility in females and males, while antimetabolites cause sterility in females. Genetic control was applied in 1954 against the gray blowfly on the island of Curaçao, which causes significant damage to livestock production. The release of sterilized individuals was successful. The genetic control method is inherently selective; its use is not associated with a negative impact on the environment and does not contribute to the development of resistance to sterilization factors.

History of biotechnology

Since ancient times, people have used biotechnological processes in baking, preparing fermented milk products, winemaking, etc., but only thanks to the work of Louis Pasteur in the mid-19th century, they proved the connection between fermentation processes and the activity of microorganisms, and traditional biotechnology received a scientific basis.

In the 40-50s of the 20th century, when the biosynthesis of penicillins was carried out using fermentation methods, the era of antibiotics began, which gave impetus to the development of microbiological synthesis and the creation of the microbiological industry.

In the 60-70s of the 20th century, cell engineering began to develop rapidly.

The creation of the first hybrid DNA molecule in vitro in 1972 by P. Berg's group in the USA was formally associated with the birth of genetic engineering, which opened the way to consciously changing the genetic structure of organisms in such a way that these organisms could produce the products necessary for humans and carry out the necessary processes. These two directions have determined the appearance of a new biotechnology that has little in common with the primitive biotechnology that man has used for thousands of years. It is significant that in the 1970s the term itself became widespread biotechnology. Since that time, biotechnology has been inextricably linked with molecular and cellular biology, molecular genetics, biochemistry and bioorganic chemistry. In a short period of its development (25-30 years), modern biotechnology has not only achieved significant success, but also demonstrated unlimited possibilities for the use of organisms and biological processes in various sectors of production and the national economy.

Biotechnology as a science

Biotechnology is a complex of fundamental and applied sciences, technical means aimed at obtaining and using cells of microorganisms, animals and plants, as well as their metabolic products: enzymes, amino acids, vitamins, antibiotics, etc.

Biotechnology, which includes industrial microbiology, is based on the use of knowledge and methods of biochemistry, microbiology, genetics and chemical technology, which allows technological processes to benefit from the properties of microorganisms and cell cultures. Modern biotechnological processes are based on recombinant DNA methods, as well as on the use of immobilized enzymes, cells and cellular organelles.

Main areas of research:

• Development of the scientific basis for the creation of new biotechnologies using the methods of molecular biology, genetic and cellular engineering.
• Production and use of microbial biomass and microbiological synthesis products.
• Study of the physicochemical and biochemical foundations of biotechnological processes.
• Using viruses to create new biotechnologies.

Application

Biotechnology is applied around us in many everyday items - from the clothes we wear to the cheese we consume. For centuries, farmers, bakers, and brewers have used traditional techniques to alter and modify plants and foods—wheat may be the oldest example, and nectarine one of the most recent. Today biotechnology uses modern scientific methods, which allow us to improve or modify plants, animals, microorganisms with greater accuracy and predictability.

Consumers should have a wider range of safe products to choose from. Biotechnology can provide consumers with these choices—not only in agriculture, but also in medicine and fuel resources.

Benefits of biotechnology

Biotechnology offers enormous potential benefits. Developed countries and developing countries alike should have a direct interest in supporting further research to enable biotechnology to realize its full potential.

Biotechnology helps the environment. By allowing farmers to reduce the use of pesticides and herbicides, first-generation biotechnology products have led to a reduction in their use in agricultural practices, and future biotechnology products are expected to provide even greater benefits. Reduced pesticide and herbicide loads mean less risk of toxic contamination of soils and groundwater. In addition, herbicides used in combination with genetically modified plants are often safer for the environment than the previous generation of herbicides they replace. Bioengineered crops also contribute to the widespread use of no-moldboard tillage, which leads to a reduction in soil fertility losses.

Biotechnology has enormous potential in the fight against hunger. Developments in biotechnology offer significant potential benefits for developing countries, where more than a billion people worldwide live in poverty and suffer from chronic hunger. By increasing yields and developing crops that are resistant to disease and drought, biotechnology can reduce food shortages for the global population, which by 2025 will be more than 800,000,000 people, an increase of 30% from today. Scientists are creating crops with new properties that help them survive the harsh conditions of drought and floods.

Biotechnology helps fight disease. By developing and improving medicine, it provides new tools in the fight against them. Biotechnology has given medical methods treatment of cardiac diseases, sclerosis, hemophilia, hepatitis, and AIDS. Biotechnological food products are now being created that will make life cheaper and more accessible for the poorest part of the planet’s population. essential vitamins and vaccines.

Cautions for use

The volume of removal of bioproducts from the biosphere has reached 70%, and living matter functions at an optimal level when no more than 15% of biosphere products are withdrawn. Ecosystems and the biosphere as a whole are increasingly losing the ability to self-regulate and self-support. Ultimately, this gives the circulation of substances on the globe a qualitatively new and unpredictable character. The stability of the functioning of the biosphere was under threat. Pollution and degradation cover all geospheres of the Earth. Air, water and soil began to lose their basic natural properties.

Biotechnology in healthcare

Biotechnology can bring significant benefits to the healthcare industry. By increasing the nutritional value of food, biotechnology can be used to improve nutritional quality. For example, varieties of rice and corn with increased protein content are now being created. In the future, consumers will be able to use oil with a reduced fat content, which will be obtained from genetically modified corn, soybeans, and rapeseed. In addition, genetic engineering could be used to produce foods with increased levels of vitamin A, which could help solve the problem of blindness in developing countries. Genetic engineering also offers other health benefits, with techniques now available that can remove certain allergenic proteins from food or prevent it from spoiling prematurely.

Biotechnology in medicine

In medicine, biotechnological techniques and methods play a major role in the creation of new biologically active substances and drugs intended for early diagnosis and treatment of various diseases. Antibiotics are the most class of pharmaceutical compounds obtained by microbiological synthesis. Genetically engineered strains of Escherichia coli, yeast, cultured mammalian and insect cells have been created, used to produce growth hormone, insulin and human interferon, various enzymes and antiviral vaccines. By changing the nucleotide sequence in the genes encoding the corresponding proteins, the structure of enzymes, hormones and antigens is optimized (the so-called protein engineering). The most important discovery was the technique developed in 1975 by G. Köhler and S. Milstein for using hybridomas to obtain monoclonal antibodies of the desired specificity. Monoclonal antibodies are used as unique reagents for the diagnosis and treatment of various diseases.

Biotechnology in agriculture

Biotechnology in agriculture facilitates traditional methods of plant and animal breeding and develops new technologies to improve agricultural efficiency. In many countries, highly productive varieties of agricultural plants that are resistant to pests, diseases, and herbicides have been created using genetic and cellular engineering methods. A developed technique for healing plants from accumulated infections, which is especially important for crops that reproduce vegetatively (potatoes, etc.). As one of the most important problems biotechnology all over the world, research into the possibility of controlling the process of nitrogen fixation, the possibility of introducing nitrogen fixation genes into the genome of useful plants, as well as the process of photosynthesis. Improvements in the amino acid composition of plant proteins are being investigated. New plant growth regulators, microbiological means of protecting plants from diseases and pests, and bacterial fertilizers are being developed. Genetically engineered vaccines, serums, and monoclonal antibodies are used for the prevention, diagnosis and treatment of major diseases in animal husbandry. To create effective breeding technologies, genetically engineered growth hormone is used, as well as transplantation and micromanipulation techniques on animal embryos. To increase animal productivity, feed protein obtained by microbiological synthesis is used.

Biotechnology in production

Biotechnological processes using microorganisms and enzymes at the modern technical level are widely used in the food industry. Industrial cultivation of microorganisms, plant and animal cells is used to obtain many valuable compounds - enzymes, hormones, amino acids, vitamins, antibiotics, methanol, organic acids (acetic, citric, lactic), etc. With the help of microorganisms, the biotransformation of some organic compounds into others is carried out ( for example, sorbitol to fructose). Immobilized enzymes are widely used in various industries. Monoclonal antibodies are used to isolate biologically active substances from complex mixtures. A. S. Spirin in 1985-1988 developed the principles of cell-free protein synthesis, when instead of cells special bioreactors containing necessary set purified cellular components. This method produces different types of proteins and can be efficient in production. Many industrial technologies are being replaced by technologies that use enzymes and microorganisms. Such biotechnological methods for processing agricultural, industrial and household waste, treating and using wastewater to produce biogas and fertilizers. In a number of countries, microorganisms are used to obtain ethanol, used as fuel for cars (in Brazil, where fuel alcohol is widely used, it is obtained from sugar cane and other plants). The extraction of many metals from low-grade ores or wastewater is based on the ability of various bacteria to transfer metals into soluble compounds or accumulate them.

Bionanotechnology

Development of biological materials and special processes that use nanomaterials or nanotechnologies. Including molecular motors, biomaterials, single molecule manipulation technology, biochip technology.

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