Molecular cloning and expression of a gene encoding a novel sorbitol oxidase from Streptomyces sp. H-7775.
A gene encoding a novel intracellular sorbitol oxidase of a soil bacterium, Streptomyces sp. H-7775, was cloned and sequenced. The gene consists of an open reading frame of 1,260-bp encoding a protein of 420 amino acids with a molecular weight of 45,148. Deduced amino acid sequence of the gene has 25.3% identity and 68.1% similarity to that of rat L-gulonolactone oxidase at the overall amino acids. Nucleotide-binding motifs were not found in the deduced amino acid sequence of SOX protein. We succeeded in expressing recombinant sorbitol oxidase with covalently bound FAD in E. coli at about a 4,000-fold higher total enzyme activity than that of the Streptomyces sp. H-7775. The enzymatic properties of the recombinant SOX were similar to those of the enzyme from Streptomyces sp. H-7775. This is the first report of the cloning and expression of a newly categorized enzyme, sorbitol oxidase, from Streptomyces sp.
Hiraga
K
K
Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Japan.
Eto
T
T
Yoshioka
I
I
Oda
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002847:Chromatography, Agarose; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D010088:Oxidoreductases; C111983:sorbitol oxidase