Z.K. Nikitina, V.A. Bykov

The review uses date obtained by the authors and available in the literature to discuss peculiarities of manipulation with microorganisms productive of biologically active substances. The main directions in selection, production gain and optimization of cultivation and conservation conditions are analyzed with deuteromyces, secreting keratinases and collagenases, and recombinant strains of Escherichia coli and Saccharomyces cerevisiae, synthesizing epidermal growth factor (EGF), as an example. Methods of keratin-containing substrates based on human hair and rat tails epidermis obtaning were developed. Micromycetes were screened with application ofl these substrates as the source of carbon for synthesis of extracellular keratinases. The highest index of keratin lysis, growth rate, the highest keratinolitic activity of culture media were found for Сladosporium sphaerospermum and Penicillum citrinum. The perspective keratinases obtaining technology from Cl. sphaerospermum and P. citrinum was developed. EGF is a promising drug for therapy. Its production from natural sources is limited by the insufficient available. The present paper discusses the feasibility for production of recombinant EGF by various microorganisms and expression systems. E. coli recombinant strain produced EGF biological properties were studied in different cultivation conditions and following results were shown: LB medium stimulates cell growth and mass better then M9 medium; IPTG inductor application procedure was developed; inductor optimal concentration was also found. S. cerevisiae recombinant strains produced EGF biological properties were studied in different cultivation conditions and optimal composition of growths medium was developed. It was found that collagen was induced synthesis of collagenolytic enzymes by deuteromycetes. It is shown that the reseedings lead to reduction of the share of plasmid cells, the growth rate and productivity of the recombinant strains E. coli. Storage at a temperature of liquid nitrogen they made it possible to preserve plasmid cells and secure the stability of biosynthetic properties. It was found that increase number passage and storage on medium with collagen increased collagenolytic activity of Aspergillus flavus.