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Journal №5 for 2014 г.
Article in number:
Penicillium citrinum keratinase obtaining method optimization by using culturе adaptation and exogenous regulation of enzyme biosynthesis
Keywords: keratinase keratinolytic activity deuteromyces Penicillium citrinum
Authors:
Z.K. Nikitina - Dr. Sc. (Biol.), Professor, All-Russian Scientific, Institute of Medicinal and Aromatic Plants, Moscow, Russia
I.K. Gordonova - Ph. D. (Biol.), Leading Research Scientist, All-Russian Scientific, Institute of Medicinal and Aromatic Plants, Moscow, Russia
Abstract:
Keratinases are the promising drugs for therapy and use in different fields were keratins should be hydrolyzed. The method of isolation and purification of keratinolytic enzyme, secreting by Penicillium citrinum PC-54-91VILAR, was developed by us earlier and biopharmaceutical properties of this ferment were also studied. The aim of this investigation was the keratinase of P. citrinum PC-54-91VILAR obtaining method improvement. P. citrinum PC-54-91VILAR strain directional adaptation by 3 times cultivation in Czapek-agar media, containing hair keratin (2 %) instead of saccharose, for its biosynthetic activity increase was conducted. In result, micromycete cultivation time was reduced from 10 to 6 days and its total keratinilytic activity was increased in 2,4 times. Subsequent cultivation times increase didn-t modificate studied parameters. Later investigations showed that hair keratin concentration in culture medium was the important factor of exogenic regulation. Increase of protein concentration from 1,5 to 10 % allowed increase adapted culture broth filtrate enzyme activity on 141%. Cultivation time didn-t change in this case. The developed method permits to get pure keratinase which can destroy hair ?keratin with 19 mg/l culture liquid productivity, 231 KE/mg enzyme activity, 70 % yield and 59,3 times degree of purification. Obtained results allow decrease the cultivation time from 10 to 6 days, increase ferment yield in 5,8 times and thus improve ferment obtaining technology for creation new drugs.
Pages: 32-36
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