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KMID : 0613820130230020273
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Journal of Life Science
2013 Volume.23 No. 2 p.273 ~ p.281
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Production and Characterization of Alkaline Protease of Micrococcus sp. PS-1 Isolated from Seawater
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Jin Young-Rang
Yu Sun-Nyoung
Kim Kwang-Youn
Kim Sang-Hun
Park Seul-Ki
Kim Hyeun-Kyeung
Lee Yong-Seok
Choi Yong-Lark
Ji Jae-Hoon
Ahn Soon-Cheol
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Abstract
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The purpose of this research was to investigate the production and characterization of alkaline protease from Micrococcus sp. PS-1 newly isolated from seawater. Micrococcus sp. PS-1 was grown in Luria-Bertani (LB) medium. Its optimal temperature and pH for growth were 30℃ and 7.0, respectively. The effect of nitrogen sources was investigated on optimal enzyme production. A high level of alkaline protease production occurred in LB broth containing 2% skimmed milk. The protease was purified in a 3-step procedure involving ultrafiltration, acetone precipitation, and dialysis. The procedure yielded a 16.43-purification fold, with a yield of 54.25%. SDS-PAGE showed that the enzyme had molecular weights of 35.0 and 37.5 kDa. Its maximum protease activity was exhibited at pH 9.0 and 37℃, and its activity was stable at pH 8.0-11.0 and 25-37℃. The protease activity was strongly inhibited by PMSF, EDTA, and EGTA. Taken together, the results demonstrate that the protease enzyme from Micrococcus sp. PS-1 probably belongs to a subclass of alkaline metallo-serine proteases.
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KEYWORD
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Alkaline protease, Micrococcus sp., seawater, characterization, skim milk
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