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Biochemical Characterization of an Extracellular Protease in Serratia proteamaculans Isolated from a Spider  

Lee Kieun (Department of Biology, Chungnam National University)
Kim Chul-Hee (Department of Biology, Chungnam National University)
Kwon Hyun-Jung (Insect Resources Laboratory, Korea Research Institute of Bioscience and Biotechnology)
Kwak Jangyul (Insect Resources Laboratory, Korea Research Institute of Bioscience and Biotechnology)
Shin Dong-Ha (Insect Biotech Co., Ltd.)
Park Doo-Sang (Insect Resources Laboratory, Korea Research Institute of Bioscience and Biotechnology)
Bae Kyung-Sook (Insect Resources Laboratory, Korea Research Institute of Bioscience and Biotechnology)
Park Ho-Yong (Insect Resources Laboratory, Korea Research Institute of Bioscience and Biotechnology)
Publication Information
Korean Journal of Microbiology / v.40, no.4, 2004 , pp. 269-274 More about this Journal
Abstract
Serratia proteamaculans isolated from the midgut of a spider formed big halos around the bacterial colonies, indicating that the bacterial strain produces an extracellular protease. Activity staining of the extracellular pro­tein fractions using zymogram also demonstrated that the major protein with an estimated molecular mass of 52 kDa contained a high proteolytic activity. The protease was purified to near electrophoretic homogeneity from the culture supernatant after filtration and ion exchange and size exclusion chromatography. The purified enzyme had a relatively high proteolytic activity between pH 6.0 and 10.0 and at broad temperature range. The proteolytic activity of the enzyme was not inhibited by phenylmethylsulfonyl fluoride but strongly inhibited by 1, 10-phenanthroline and EDTA. The activity also was dependent on the presence of $Ca^{++}\;and\;Zn^{++}$ ions. These observations indicate that the enzyme is a metalloprotease.
Keywords
chromatography; metalloprotease; proteolytic activity; Serratia proteamaculans; zymogram;
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