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Cloning and sequence analysis of aStreptomyces cholesterol esterase gene

  • Applied Genetics and Regulation
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Abstract

Streptomyces lavendulae H646-SY2 produces cholesterol esterase (CHE; EC 3.1.1.13) extracellularly. A genomic library of the strain, prepared in plasmid pUC119, was screened with probes based on the amino acid sequence of the protein. A plasmid, designated as pKX101 and identified by hydridization with the probes, contained a 2.7-kb insert fromStreptomyces DNA. We determined the 17-N-terminal amino acid sequence of mature CHE and the nucleotide sequence of the 0.9-kb segment containing the CHE gene (che). We found that the N-terminal of the mature CHE was Ala39 and an open reading frame consisting of 681 bp starts at ATG and ends at TGA, suggesting that a precursor and a mature CHE consist of 227 and 189 amino acids, with a calculated relative molecular mass of 24,362 and 20,650, respectively. The leader peptide extends over 38 amino acids and has the characteristics of a signal sequence, including basic amino acids near the N-terminus and a hydrophobic core near the signal cleavage site.

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  1. M. Nishimura

    Present address: Department of Chemical and Biological Engineering, Ube National College of Technology, 755, Ube, Tokiwadai, Japan

Authors and Affiliations

  1. Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Kasumi 1-2-3, Minami-ku, 734, Hiroshima, Japan

    M. Nishimura & M. Sugiyama

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  1. M. Nishimura
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  2. M. Sugiyama
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Nishimura, M., Sugiyama, M. Cloning and sequence analysis of aStreptomyces cholesterol esterase gene. Appl Microbiol Biotechnol 41, 419–424 (1994). https://doi.org/10.1007/BF00939030

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  • DOI: https://doi.org/10.1007/BF00939030

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