Purification and Characterization of Serine Proteinase 2 from Bacillus intermedius 3-19 |
| |
Authors: | N P Balaban A M Mardanova M R Sharipova L A Gabdrakhmanova E A Sokolova G N Rudenskaya I B Leshchinskaya |
| |
Institution: | Department of Microbiology, Kazan State University, Kazan 420008, Russia. Nelly.Balaban@ksu.ru |
| |
Abstract: | A proteinase secreted in the late stationary phase was isolated from the culture fluid of Bacillus intermedius 3-19 by ion-exchange chromatography on CM-cellulose followed by FPLC on a Mono S column. The enzyme was completely inhibited by the serine proteinase inhibitors diisopropyl fluorophosphate and phenylmethylsulfonyl fluoride. The maximum proteolytic activity against the synthetic chromogenic substrate Z-Ala-Ala-Leu-pNA was observed at pH 9.0. The molecular weight of the enzyme is 28 kD and its isoelectric point is 9.2. We have also determined pH- and thermostability and Km and kcat of this proteinase. The enzyme has been classified as a thiol-dependent serine proteinase. N-Terminal amino acid sequence (10 residues) and amino acid composition of the protein were also determined. By the mode of hydrolysis of peptide bonds in the oxidized B-chain of insulin, this enzyme is similar to the thiol-dependent serine proteinase 1 from B. intermedius 3-19 secreted during vegetative growth. |
| |
Keywords: | proteinase thiol-dependent serine proteinase (Bacillus intermedius) purification properties |
本文献已被 PubMed SpringerLink 等数据库收录! |