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Cloning, nucleotide sequence, and overexpression in Escherichia coli of the β-tyrosinase gene from an obligately symbiotic thermophile, Symbiobacterium thermophilum

  • Applied Genetics and Regulation
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Abstract

Symbiobacterium thermophilum is an obligately symbiotic thermophile that can grow only in coculture with a specific Bacillus strain. The amino acid sequences of fragments obtained by cyanogen bromide decomposition of the thermostable β-tyrosinase (tyrosine phenol-lyase, E.C. 4.1.99.2) from this organism resembled that of the tryptophanase produced by the same organism. DNA-probing with the tryptophanase gene as the hybridization probe led to cloning in Escherichia coli of the β-tyrosinase (tpl) gene. The nucleotide sequence revealed that the β-tyrosinase of 458 amino acids (relative molecular mass, 52269) showed significant similarity in amino acid sequence to the tryptophanase over the entire sequence. DNA manipulation of the cloned tpl gene in E. coli led to production of 375 times as much β-tyrosinase as that produced by the original S. thermophilum strain.

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Authors and Affiliations

  1. Department of Agricultural Chemistry, Faculty of Agriculture, The University of Tokyo, Bunkyo-ku, 113, Tokyo, Japan

    Toshikatsu Hirahara, Sueharu Horinouchi & Teruhiko Beppu

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  1. Toshikatsu Hirahara
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  2. Sueharu Horinouchi
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  3. Teruhiko Beppu
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Hirahara, T., Horinouchi, S. & Beppu, T. Cloning, nucleotide sequence, and overexpression in Escherichia coli of the β-tyrosinase gene from an obligately symbiotic thermophile, Symbiobacterium thermophilum . Appl Microbiol Biotechnol 39, 341–346 (1993). https://doi.org/10.1007/BF00192089

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  • DOI: https://doi.org/10.1007/BF00192089

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