Cloning, nucleotide sequence, and overexpression inEscherichia coliof the β-tyrosinase gene from an obligately symbiotic thermophile,Symbiobacterium thermophilum
✍ Scribed by Toshikatsu Hirahara; Sueharu Horinouchi; Teruhiko Beppu
- Publisher
- Springer
- Year
- 1993
- Tongue
- English
- Weight
- 857 KB
- Volume
- 39
- Category
- Article
- ISSN
- 1432-0614
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✦ Synopsis
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 fl-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 fl-tyrosinase (tpl) gene. The nucleotide sequence revealed that the fl-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 fl-tyrosinase as that produced by the original S. thermophilum strain.