미생물학회지 (Korean Journal of Microbiology)
- 제30권6호
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- Pages.507-513
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- 1992
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- 0440-2413(pISSN)
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- 2383-9902(eISSN)
Cloning, Sequencing and Expression of an Extracellular Protease Gene from Serratia marcescens RH1 in Escherichia coli
- Lee, Seung-Hwan (Department of Molecular Biology, Seoul National University) ;
- Kim, Jeong-Min (Department of Molecular Biology, Seoul National University) ;
- Kwon, Young-Tae (Department of Molecular Biology, Seoul National University) ;
- Kho, Young-Hee (Genetic Engineering Research Institute, KAIST) ;
- Rho, Hyune-Mo (Department of Molecular Biology, Seoul National University)
- 발행 : 1992.12.01
초록
Serratia marecescens RH1 isolated from soil samples produced large amount of extracellular proteases. One of the genes encoding an extracellular protease form S. marcescens RH1 was cloned in Escherichia coli by shot gun cloning method. The cloned protease, SSP, was stably expressed by its own promoter and excreted into the extracellular medium from E. coli host (ORF) of 3.135 nucleotides corresponding to 1.045 amino acids (112 kDa). The nucleotide and deduced amino acid sequence of SSP showed high overall homology (88%) to one of the S. marcescens protease (27), but low homology to other serine protease families. The optimal pH and temperature of the enzyme were pH 9.0 and 45.deg.C respectively. The activity of protease was inhibited by phenylmethylsulfonyl fluoride (PMSF), which suggests that the enzyme is a serine protease.