Cloning and expression of the sarcosine oxidase gene from Bacillus sp. NS-129 in Escherichia coli.
The gene coding for a thermostable sarcosine oxidase (EC 1.5.3.1) was isolated from Bacillus sp. NS-129. The primary structure of sarcosine oxidase deduced from the nucleotide sequence was a protein composed of 387 amino acids with molecular weight 42,955. When the sarcosine oxidase was overproduced to about 35% of soluble protein in E. coli under the control of a lac promoter, the sarcosine oxidase activity of the crude extract was increased 3-fold by the addition of FAD. This indicates that most of the enzyme is accumulated in an active form, a flavinless aporotein, in the cell.
Koyama
Y
Y
Research and Development Division, Kikkoman Corporation, Chiba, Japan.
Yamamoto-Otake
H
H
Suzuki
M
M
Nakano
E
E
D000595:Amino Acid Sequence; D001407:Bacillus / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010089:Oxidoreductases, N-Demethylating / Q000235:genetics*; D050879:Sarcosine Oxidase
D010089:Oxidoreductases, N-Demethylating; D050879:Sarcosine Oxidase