Abstract
In this study, a 107 kDa protease from psychrophilic Janthinobacterium lividum PAMC 26541 was purified by anion-exchange chromatography. The specific activity of the purified protease was 264 U/mg, and the overall yield was 12.5%. The J. lividum PAMC 25641 protease showed optimal activity at pH 7.0-7.5 and 40°C. Protease activity was inhibited by PMSF, but not by DTT. On the basis of the N-terminal sequence of the purified protease, the gene encoding the cold-adapted protease from J. lividum PAMC 25641 was cloned into the pET-28a(+) vector and heterologously expressed in Escherichia coli BL21(DE3) as an intracellular soluble protein.
Keywords:
Cold-adapted protease; Janthinobacterium lividum; expression; purification.
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism
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Cloning, Molecular
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Cold Temperature*
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DNA, Bacterial
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Enzyme Activation
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Enzyme Stability
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Escherichia coli / genetics
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Gene Expression Regulation, Enzymologic
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Hydrogen-Ion Concentration
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Metals
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Molecular Weight
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Oxalobacteraceae / enzymology*
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Oxalobacteraceae / genetics*
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Peptide Hydrolases / chemistry*
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Peptide Hydrolases / genetics*
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Peptide Hydrolases / isolation & purification*
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Peptide Hydrolases / metabolism*
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Substrate Specificity
Substances
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Bacterial Proteins
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DNA, Bacterial
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Metals
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Recombinant Proteins
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Peptide Hydrolases