Cloning of the biotin synthetase gene from Bacillus sphaericus and expression in Escherichia coli and Bacilli.
Biotin synthetase (BS) catalyses the biotransformation of dethiobiotin (DTB) to biotin. Here we report the cloning, characterization and expression of the gene encoding BS of Bacillus sphaericus. A recombinant plasmid pSB01, containing an 8.2-kb DNA fragment from B. sphaericus, was isolated by phenotypic complementation of an Escherichia coli bioB strain. Nucleotide sequence analysis of this fragment and N-terminal sequence determination of the recombinant protein product revealed that the bioB gene of B. sphaericus consists of a 996-bp open reading frame which is closely associated with at least one other gene. E. coli cells transformed with a bioB expression vector performed efficient bioconversion of DTB to biotin under defined culture conditions. Biotin production from transformed Bacillus subtilis and B. sphaericus recombinant strains was also demonstrated. Comparison of the amino acid sequences of BS from E. coli and B. sphaericus revealed extensive similarity.
Ohsawa
I
I
Biological Science Institute, R & D Center, Nippon Zeon Co., Ltd., Kawasaki, Japan.
Speck
D
D
Kisou
T
T
Hayakawa
K
K
Zinsius
M
M
Gloeckler
R
R
Lemoine
Y
Y
Kamogawa
K
K
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D001710:Biotin / Q000096:biosynthesis; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005784:Gene Amplification; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D013466:Sulfurtransferases / Q000096:biosynthesis / Q000235:genetics*; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D001710:Biotin; D013466:Sulfurtransferases; C057985:biotin synthetase