Cloning, Heterologous Expression and Characterization of an Intracellular Serine Protease from Bacillus sp. LCB10

An intracellular serine protease designated ISPr from newly isolated salt-tolerant Bacillus sp. LCB10 was expressed in Escherichia coli . The isp gene was cloned into pET30a vector and expressed in E. coli BL21 (DE3). After purification, the molecular mass of ISPr was estimated to be 35 kDa by SDS-P...

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Bibliographic Details
Published in:Applied biochemistry and microbiology 2019-09, Vol.55 (5), p.482-488
Main Authors: Hou, Y., Lu, F., Tian, J., Tian, Y.
Format: Article
Language:English
Subjects:
Bacillus
Biochemistry
Biomedical and Life Sciences
Cloning
E coli
Enzymes
Gel electrophoresis
Intracellular
Life Sciences
Medical Microbiology
Microbiology
pH effects
Protease
Purification
Salinity tolerance
Serine
Serine proteinase
Sodium chloride
Sodium lauryl sulfate
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Summary:An intracellular serine protease designated ISPr from newly isolated salt-tolerant Bacillus sp. LCB10 was expressed in Escherichia coli . The isp gene was cloned into pET30a vector and expressed in E. coli BL21 (DE3). After purification, the molecular mass of ISPr was estimated to be 35 kDa by SDS-PAGE and the specific activity was 384 U/mg. The protease showed optimal activity at 40°C and pH 10.0. ISPr was active and stable in wide range of alkaline pH. ISPr activity increased 1.8-fold in the presence of Mn 2+ and was completely inhibited by PMSF. The enzyme exhibited high NaCl tolerance. The protease had a stable activity in the presence of 1–5% NaCl and retained 86% activity in the presence 7% NaCl, thus, this enzyme would have great potential in industry.
ISSN:0003-6838
1608-3024
DOI:10.1134/S0003683819050168