The thesis for a candidate's degree of Biological Science by speciality 03.00.07 - microbiology. - Institute of microbiology and virology named D.K. Zabolotny of National Academy of Sciences of Ukraine, Kiev, 2009. The dissertation is focused on purification and characterization of proteolytic complex of Вacillus sp. 27, Вacillus circulans 693 and Yarrowia lipolytica 2061. As a result of screening executed on the basis of 367 strains of microorganisms - yeast (Debaryomyces, Kluyveromyces, Saccharomyces, Yarrowia) and bacteria (Bacillus) there have been selected three strains-producer of proteolytic enzymes Вacillus sp. 27, В. circulans 693 and Y. lipolytica 2061 with the highest level of activity. The most appropriate souses of carbon and nitrogen, respectively, for Вacillus sp. 27 were arabinose and gelatin, for В. circulans 693 - glucose and the combination of gelatin with the ammonium hydrocarbonate, for Y. lipolytica 2061 - sorbose and the gelatin with glycine. This gave a possibility to increase the proteolytic activity of Вacillus sp. 27, В. circulans 693 and Y. lipolytica 2061 in 2,7; 3,8 and correspondingly in 3,4 times. The usage of substances of natural or synthetic origin as the inductors testifies that the bovine blood, elastin, gelatin, haemoglobin, casein and some coordination compounds of germanium (in particular, the complexes of germanium with aspartic acid or methionine), caused the increase of enzyme biosynthesis in 2,3 - 5,8 times. The most promising turned out to be five fractions: 1(1) and 2(2) Bacillus sp. 27, which demonstrated a high level of fibrinolytic and elastase activity, respectively; 1(1) В. circulans 693 with dominating haemoglobinolitic and fibrinolytic activities, 1(1) and 1(3) Y. lipolytica 2061, which were characterized by high levels of fibrinolytic and gelatinase activities, respectively. Hereby, the following preparations were obtained: three with fibrinolytic activity, one with haemoglobinolitic activity, and other two showed the gelatinolitic and elastase activities. As a result, the specific activity of enzymes was 11,8; 22 and 33 times increased for the preparations with fibrinolytic activity; for the preparation with hemoglobinolitic activity it was raised to 22,4 times; for the one with the gelatinolitic activity to 27 times, with the elastase activity - to 40 times. The protease preparations of all the three producers: Вacillus sp. 27, В. circulans 693, and Y. lipolytica 2061 belong to the group of serine proteases, which, probably, for the manifestation of their activity need to be influenced by the ions of metals, and contain the residues of cysteine close to their active center.
