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Characterization of proteolytic bacteria from the Aleutian deep-sea and their proteases

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  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Six deep-sea proteolytic bacteria taken from Aleutian margin sediments were screened; one of them produced a cold-adapted neutral halophilic protease. These bacteria belong to Pseudoalteromonas spp., which were identified by the 16S rDNA sequence. Of the six proteases produced, two were neutral cold-adapted proteases that showed their optimal activity at pH 7–8 and at temperature close to 35°C, and the other four were alkaline proteases that showed their optimal activity at pH 9 and at temperature of 40–45°C. The neutral cold-adapted protease E1 showed its optimal activity at a sodium chloride concentration of 2 M, whereas the activity of the other five proteases decreased at elevated sodium chloride concentrations. Protease E1 was purified to electrophoretic homogeneity and its molecular mass was 34 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of protease E1 was determined to be 32,411 Da by mass spectrometric analysis. Phenylmethyl sulfonylfluoride (PMSF) did not inhibit the activity of this protease, whereas it was partially inhibited by ethylenediaminetetra-acetic acid sodium salt (EDTA-Na). De novo amino acid sequencing proved protease E1 to be a novel protein.

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Acknowledgments

We thank the Mass Spectrometry Facility at the Hong Kong University of Science and Technology for the ESI mass spectrometry and the de novo protein-sequencing analysis. We thank Drs. Hans-U. Dahms and J. R. Wu for their critical comments, and Dr. Mike Poole and Mr. Drew Wilson for their editorial work on the manuscript. This work was supported by a grant (COMRRDA 03/04 SC01) from the China Ocean Mineral Resources Research and Development Association to PY Qian.

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Authors and Affiliations

  1. Coastal Marine Laboratory, Department of Biology, Hong Kong University of Science and Technology, Kowloon, Hong Kong SAR, China

    Hairong Xiong, Ying Xu, Man-Yee Tsoi, Sergey Dobretsov & Pei-Yuan Qian

  2. Experimental Marine Biology Laboratory, Institute of Oceanology, Chinese Academy of Sciences, Qindao, 266071, China

    Linsheng Song

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  1. Hairong Xiong
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  2. Linsheng Song
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Correspondence to Pei-Yuan Qian.

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Xiong, H., Song, L., Xu, Y. et al. Characterization of proteolytic bacteria from the Aleutian deep-sea and their proteases. J Ind Microbiol Biotechnol 34, 63–71 (2007). https://doi.org/10.1007/s10295-006-0165-5

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  • DOI: https://doi.org/10.1007/s10295-006-0165-5

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