Abstract
The gene kerA (1,047 bp) encoding the main keratinase from Bacillus licheniformis was cloned into two conventional vectors, pET30α and pET32α, and expressed in Escherichia coli. From SDS-PAGE analysis, the recombinant keratinases were 45 and 55 kDa. They had different optimal pH values (7.5 and 8.5) but the same optimum temperature of 50 °C. The recombinant keratinase produced in E. coli pET30α-kerA was more stable than that produced in E. coli pET32α-kerA, and retained approx. 70 % of its total enzyme activity after 30 min at 70 °C.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacillus / enzymology*
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Bacillus / genetics
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Cloning, Molecular
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DNA, Bacterial / chemistry
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DNA, Bacterial / genetics
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Electrophoresis, Polyacrylamide Gel
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Enzyme Stability
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Escherichia coli / genetics
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Gene Expression
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Hydrogen-Ion Concentration
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Molecular Sequence Data
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Molecular Weight
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Peptide Hydrolases / chemistry
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Peptide Hydrolases / genetics*
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Peptide Hydrolases / metabolism*
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Analysis, DNA
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Temperature
Substances
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DNA, Bacterial
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Recombinant Proteins
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Peptide Hydrolases
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keratinase