Abstract
An extracellular protease was purified from a deep-sea psychrophilic bacterium strain DY-A which was identified as a Pseudomonas species. The optimal growth and protease-producing temperatures of the strain were all 10°C, and the protease was secreted only at temperatures under 20°C. The enzyme was most active at 40°C and at pH 10.0. It was inhibited by phenylmethyl sulfonylfluoride and diisopropyl fluorophosphate, indicating that it is a serine protease. Chelators such as EDTA, EGTA, 1,10-phenanthroline and 2,2′-bipyridyl produced a decrease of activity. The enzyme was sensitive to denaturing agents such as SDS, urea, and guanidine HCl and resistant to thiol-containing reducing agents such as dithiotreitol. The enzyme was active towards N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide and N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide. The native molecular mass of the enzyme determined by native PAGE and SDS-PAGE was 25 kDa.
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Acknowledgements
This work was supported by NKBRSF Project G2000078500 and COMRA Project DY105-4-2-4.
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Communicated by K. Horikosui
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Zeng, R., Zhang, R., Zhao, J. et al. Cold-active serine alkaline protease from the psychrophilic bacterium Pseudomonas strain DY-A: enzyme purification and characterization. Extremophiles 7, 335–337 (2003). https://doi.org/10.1007/s00792-003-0323-x
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DOI: https://doi.org/10.1007/s00792-003-0323-x