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Characterization of an alkaline serine protease from an alkaline-resistant Pseudomonas sp.: Cloning and expression of the protease gene in Escherichia coli

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Summary

A gene, aprP, encoding an extracellular alkaline serine protease from a newly isolated Pseudomonas sp. KFCC 10818 was cloned and characterized. Nucleotide sequence analysis revealed an open reading frame of 1,266 nucleotides which could encode a polypeptide comprised of 422 amino acids. The C-terminal 283 residues showed an overall sequence homology with the subtilisin-type serine proteases. When expressed in E. coli, the alkaline protease, AprP, was released to the culture medium. The purified AprP was most active at pH 11. The k Cat/K m value of this enzyme was 9.2 × 103 S−1mM−1, which is much higher than those of subtilisins.

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Authors and Affiliations

  1. Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1 Kusong-Dong, 305-701, Yusong-Gu, Taejon, Korea

    Won Hee Jang, Eun Kyung Kim, Jae Hoon Chung & Ook Joon Yoo

  2. Department of Biology, College of Natural Sciences, Chonnam National University, 300 Yongbong-dong, 500-757, Kwangju, Korea

    Hwanghee Blaise Lee

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  1. Won Hee Jang
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  2. Eun Kyung Kim
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  3. Hwanghee Blaise Lee
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  4. Jae Hoon Chung
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  5. Ook Joon Yoo
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Jang, W.H., Kim, E.K., Lee, H.B. et al. Characterization of an alkaline serine protease from an alkaline-resistant Pseudomonas sp.: Cloning and expression of the protease gene in Escherichia coli . Biotechnol Lett 18, 57–62 (1996). https://doi.org/10.1007/BF00137811

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  • DOI: https://doi.org/10.1007/BF00137811

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