Biotechnology Basics 1400-114PBT

The content of lecture: Introduction to biotechnology, the definition of term the biotechnology and the historical outline of object, literature of subject. The perspective of development of biotechnology - the statistical connected with biotechnology data, biotechnology 21 age, social the and legal aspects the development of biotechnology (the present legal state, GMO, working political, the mistake of relinquishment, the consumer's privileges, the list of potential threats, perspective, the directions of development and the specific elements of development).The methodology of working out the biotechnological process: the modernization and the innovation of technological process, creating the new biotechnologies, preliminary phase, research, initiating the process, stages of working out the biosynthesis (selection suitable micro-organisms, preliminary decision about conditions of culturing, improvement the features of productive strains, optimization of the bioprocessing, enlarging the scale of process, starting the production). The micro-organisms in biotechnology (the review of the most important processes, formation the course of the biotechnological process, selection of strain, leadership of culture, storage the strains, the bioprocessing technology ( the led by micro-organisms fermentation, technology of recombinant DNA). The bacteriophages and bacteria in biotechnology - the bacteria genetics and the biotechnology, basis of genetic engineering and the bio-nanotechnology. The kinds of cloning - molecular and cellular.The types of cultures - the discussion the most important types of culturing micro-organisms and their meaning in production. The search the new secondary metabolite - the biological and chemical screening the antibiotics, use of high pressure and gas chromatography in process of analysis of the post-culturing products, isolation and the cleaning the new antibiotics. The use of biotechnology in medicine, agriculture, food industry, pharmaceutical industry and the protection the environment.
The mechanisms of pathogenesis and the production of the vaccines - use of alive vectors in biotechnology of vaccines of the newest generation (the induction of the intracellular immunity, acquaintance of molecular mechanisms of toxins activity and different determinants of pathogenesis in projecting the vaccines of the newest generation, bacterial ghosts) and also the use against cancer. The review of basic mechanisms of modification negative control metabolism relationships, change of metabolic preferences, activity of enzymes, control by the precursors' addition, the utilization the auxtrofic mutants and the regulatory mutants also the change penetrabilities of cellular membrane. The review of basic biochemical technologies. The overproduction of citric acid as well as the lizyne and the glutaminians. Secondary metabolites (idiolites), their biosynthesis pathway ( the ketonic route, the mevalonate route, Rohmers way, biosynthesis of peptide amino acids). The specific mechanisms of control of biosynthesis the idiolites ( the catabolic repression, control by compounds of nitrogen, the compounds of phosphorus or the energetical level in cell). Using the microorganisms and the enzymes to transformation of compounds ( the distributing the reacemic mixtures, the fermentations, the biotransformation of antibiotics, sterides). Hydrobiometalurgy. The utilization the microorganisms to lixiviation of metals ( the iron, uranium), as well as their concentration and the accumulation.

The practices program covers issues closely related to the program of lectures.
For the proposed topics include: Introduction to basic techniques and methods used in biotechnology and presentation of application of microbiology in medical and industrial biotechnology. Construction, culturing and transgenic microorganisms' profile which can have the use near projecting the vaccines of new generation. The acquaintance of molecular mechanisms of toxins activity and different determinants of pathogenesis in projecting the vaccines of the newest generation - realized in practice as a marking the activity of bacterial toxins, qualification the cytotoxicity of chosen factors of pathogenesis of Listeria monocytogenes on eukaryotic cells model, isolation of the chromosomal DNA and amplification of the genes coding the determinants of pathogenesis by method of reaction PCR. This practice subject issue is also addressed in the course of the application of bacterial pathogen proteins in medicine by demonstrating the activity of streptokinase of Streptococcus equisimilis produced by recombinant Escherichia coli strains. The silencing of bacterial gene expression at the RNA level enables a comprehensive modification of microorganisms and creates new opportunities for the control of pathogenic bacteria. This topic is realized during the course which enables to learn the principles of designing artificial RNA on the example of atsRNA (artificial trans-encoded small RNA) and evaluate the effectiveness of silencing of expression of E. coli essential genes by atsRNA. Biosurfactants are an alternative to synthetic surfactants. Due to their properties, biosurfactants can be practically used in the pharmaceutical and cosmetic industries and can also be applied in bioremediation of soil from petroleum products and heavy metals. The practical course allows the identification of bacterial strains able to produce biosurfactants, and assess whether these compounds are produced constitutively or their production requires the presence of an inducer. In addition, the ability of the tested strains for the production of emulsifiers is also studied. The utilization the simple methods to cleaning and isolating compounds: the cleaning the compounds by the extraction method connected with change pH (to isolating from hydrolysate of vegetable oil the fatty acids, the glicoalcaloids from potato); the method of isolating from fresh material the lipids, using the adsorption chromatography ( the glicolipids, the steroids, the isoprenoid dyes); the use of methods of selective precipitaiting and the extraction to receiving the phospholipids, free the and connected as a glycoside to oleanic acid oleanolowego and antocyans); distinguishing polysaccharides (starch and her components, pectins). The use affinity chromatography for purification of proteins from complex mixtures: purification of IgG from rabbit serum on Protein A-Sepharose column. Progress of purification is monitored by SDS-PAGE after Laemmli. Apparent molecular mass of the purified proteins is calculated using inverse-log formula for mobility in electrophoresis, with the use of protein molecular mass standards. The utilization of immobilized cells for enzymatic reactions in biofermentors: baker’s yeast immobilized in calcium alginate are used to convert glucose into ethanol in a flow-through microreactor. Kinetics of ethanol formation is monitored by enzymatic reaction coupled with reduction of NAD+; NADH formed is determined spectrometrically.