Molecular cloning and expression of a Streptomyces sarcosine oxidase gene in Streptomyces lividans.
A genomic library of Streptomyces sp. KB210-8SY, prepared in the plasmid vector pACYC184, was screened to obtain the gene encoding sarcosine oxidase with probes based on the amino acid sequence of the protein. A plasmid pSOXS13, which was isolated from a clone identified by hybridization with the probes, contained a 8.4-kb insert of Streptomyces DNA. When the 2.0-kb MIuI/EcoRV DNA fragment of pSOXS13 was inserted into the Streptomyces vector pIJ680 and introduced into S. lividans, the transformants produced 100-fold more sarcosine oxidase intracellularly than KB210-8SY. The nucleotides of the 1.7-kb fragment containing the sarcosine oxidase gene were sequenced. An open reading frame encoded a mature sarcosine oxidase consisting of 388 amino acids, with a calculated molecular mass of 42,107 daltons.
Suzuki
K
K
Research Laboratories, Toyo Jozo Co., Ltd., Shizuoka, Japan.
Ogishima
M
M
Sugiyama
M
M
Inouye
Y
Y
Nakamura
S
S
Imamura
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000378:metabolism; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D050879:Sarcosine Oxidase; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*
D003470:Culture Media; D009841:Oligonucleotides; D010089:Oxidoreductases, N-Demethylating; D050879:Sarcosine Oxidase