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A new alkaline serine protease from alkalophilic Bacillus sp.: Cloning, sequencing, and characterization of an intracellular protease

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Abstract

To obtain a new serine protease from alkalophilic Bacillus sp. NKS-21, shotgun cloning was carried out. As a result, a new protease gene was obtained. It encoded an intracellular serine protease (ISP-1) in which there was no signal sequence. The molecular weight was 34,624. The protease showed about 50% homology with those of intracellular serine proteases (ISP-1) from Bacillus subtilis, B. polymyxa, and alkalophilic Bacillus sp. No. 221. The amino acid residues that form the catalytic triad, Ser, His and Asp, were completely conserved in comparison with subtilisins (the extracellular proteases from Bacillus). The cloned intracellular protease was expressed in Escherichia coli, and its purification and characterization were carried out. The enzyme showed stability under alkaline condition at pH 10 and tolerance to surfactants. The cloned ISP-1 digested well nucleoproteins, clupein and salmin, for the substrates.

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Authors and Affiliations

  1. Laboratory of Molecular Enzymology, Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, 1-1, Tsuysumidori-Amamiyamchi, Aoba-ku, 981, Sendai, Japan

    Youhei Yamagata & Eiji Ichishima

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  1. Youhei Yamagata
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  2. Eiji Ichishima
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Additional information

The nucleotide sequence data reported in this paper will appear in the GSDB, DDBJ, EMBL, and NCBI nucleotide sequence databases with the accession number D37921.

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Yamagata, Y., Ichishima, E. A new alkaline serine protease from alkalophilic Bacillus sp.: Cloning, sequencing, and characterization of an intracellular protease. Current Microbiology 30, 357–366 (1995). https://doi.org/10.1007/BF00369863

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