CLADOSPORIUM SPHAEROSPERMUM KERATINASE BIOSYNTHESIS REGULATION

Issue: 
8
Year: 
2015

I.K. Gordonova
Ph.D. (Biol.), Leading Research Scientist, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow)
E-mail: igordonova777@yandex.ru
Z.K. Nikitina
Dr.Sc. (Biol.), Professor, Chief Research Scientist, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow)
E-mail: nikitinaz@yandex.ru

Biothechnology gives us an opportunity to get many biological active substances, uncluding keratinases. 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 Cladosporium sphaerospermum Cs-56-2010VILAR, was developed by us earlier and biopharmaceutical properties of this ferment were also studied. The aim of this investigation was the study of the Cl. sphaerospermum Cs-56-2010VILAR keratinase regulation mechanisms for enzyme obtaining method improvement. Cl. sphaerospermum Cs-56-2010VILAR strain directional adaptation by 4 times cultivation in Czapek-agar media, containing hair keratin (2%) instead of sucrose, for its biosynthetic activity increase was conducted. In result, cultivation time was reduced from 14 to 7 days and its total keratinilytic activity was increased in 1,9 times. 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 130%. Cultivation time didn’t change in this case. The developed method permits to get pure keratinase which can destroy hair α–keratin with 20 mg/l culture liquid productivity, 671,2 μM/(mg х h) enzyme activity, 71% yield and 65,8 times degree of purification. Obtained results allow decrease the cultivation time from 14 to 7 days, increase ferment yield in 4,37 times and thus improve ferment obtaining technology for creation new drugs.

Keywords: 
Keywords: keratinase
keratinolytic activity
deuteromyces
Cladosporium sphaerospermum.