Molecular cloning and expression of a Streptomyces sarcosine oxidase gene in Streptomyces lividans

• Published in: Bioscience, biotechnology, and biochemistry Vol. 56; no. 3; pp. 432 - 436
• Main Authors: Suzuki, K. (Toyo Jozo Co. Ltd., Ohito, Shizuoka (Japan)), Ogishima, M, Sugiyama, M, Inouye, Y, Nakamura, S, Imamura, S
• Format: Journal Article
• Language: English
• Published: Tokyo Taylor & Francis 1992; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Amino Acid Sequence; Bacteriology; Base Sequence; Biological and medical sciences; Biotechnology; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; cloning vectors; Cloning, Molecular; Culture Media; EXPRESION GENICA; EXPRESSION DES GENES; Fundamental and applied biological sciences. Psychology; GENE EXPRESSION; Gene Expression Regulation, Bacterial; Gene Library; gene mapping; Genetic engineering; Genetic technics; Genetics; Methods. Procedures. Technologies; Microbiology; MOLECULAR CLONING; Molecular Sequence Data; Molecular Weight; nucleotide sequence; Oligonucleotides - chemical synthesis; Oligonucleotides - metabolism; OXIDOREDUCTASES; Oxidoreductases, N-Demethylating - biosynthesis; Oxidoreductases, N-Demethylating - genetics; OXIDORREDUCTASAS; OXYDOREDUCTASE; Plasmids; SARCOSINA; SARCOSINE; Sarcosine Oxidase; STREPTOMYCES; Streptomyces - enzymology; Streptomyces - genetics; Streptomyces lividans; transformation; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Streptomycetaceae; Nucleotide sequence; Enzyme; Streptomyces; Gene expression; Sarcosine oxidase; Actinomycetales; Plasmid; Gene; Restriction map; Streptomyces lividans; Bacteria; Actinomycetes; Molecular cloning; Aminoacid sequence
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.56.432

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.