Cloning, nucleotide sequence, and overexpression in Escherichia coli of the beta-tyrosinase gene from an obligately symbiotic thermophile, Symbiobacterium thermophilum.
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 beta-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 beta-tyrosinase (tpl) gene. The nucleotide sequence revealed that the beta-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 beta-tyrosinase as that produced by the original S. thermophilum strain.
Hirahara
T
T
Department of Agricultural Chemistry, Faculty of Agriculture, University of Tokyo, Japan.
Horinouchi
S
S
Beppu
T
T
D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D001709:Biotechnology; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D013559:Symbiosis; D013696:Temperature; D014447:Tyrosine Phenol-Lyase / Q000235:genetics*
D004269:DNA, Bacterial; D014447:Tyrosine Phenol-Lyase