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Isolation and characterization of a novel oxidant‐ and surfactant‐stable extracellular alkaline protease from E xiguobacterium profundum BK ‐ P 23
Author(s) -
Anbu Periasamy,
Hur Byung Ki,
Gyun Lee Choul
Publication year - 2013
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1059
Subject(s) - phenylmethylsulfonyl fluoride , chemistry , oxidizing agent , enzyme , chromatography , protease , serine protease , pmsf , pulmonary surfactant , enzyme assay , nuclear chemistry , biochemistry , organic chemistry
The novel protease from E xiguobacterium profundum BK ‐P23 was partially purified by ammonium sulfate precipitation and further purified M ono Q 5/50 and S uperdex 200 10/300 column chromatography. The enzyme was purified 10.23‐fold with a yield of 14%. The molecular weight was estimated to be 52 kDa by SDS ‐ PAGE . The enzyme was most active at a p H of 8.0 and temperature of 40°C and the enzyme was stable between a pH of 7 and 10 and up to a temperature of 50°C. The enzyme activity was enhanced by CaCl 2 but was slightly inhibited by C o C l 2 , M g SO 4 , and A g NO 3 . In addition, this enzyme was completely inhibited by phenylmethylsulfonyl fluoride, indicating that this enzyme was a serine protease. Furthermore, the alkaline protease was more stable in the presence of surfactants such as T riton X ‐100, which was followed by T ween 80 and SDS . Moreover, the enzyme was highly stable in the presence of 1% oxidizing agent ( H 2 O 2 ). The enzyme also has significant stability (70%–80%) in a few organic solvents. Thus, the increased stability of the enzyme in the presence of oxidizing agent, surfactants, and organic solvents may find potential applications in the detergent industry and peptide synthesis in nonaqueous media.