Identification and cloning of the gene involved in the final step of chlortetracycline biosynthesis in Streptomyces aureofaciens

• Published in: Bioscience, biotechnology, and biochemistry Vol. 68; no. 6; pp. 1345 - 1352
• Main Authors: Nakano, T. (Kyowa Hakko Kogyo Co. Ltd., Machida, Tokyo (Japan). Tokyo Research Lab.), Miyake, K, Endo, H, Dairi, T, Mizukami, T, Katsumata, R
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.06.2004; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: amino acid sequences; antibacterial properties; ANTIBIOTIC PROPERTIES; Base Sequence; Biological and medical sciences; BIOSYNTHESIS; biotechnology; chlortetracycline; Chlortetracycline - biosynthesis; Cloning, Molecular; coenzyme F; DNA; Fundamental and applied biological sciences. Psychology; GENES; IDENTIFICATION; MOLECULAR CLONING; Molecular Sequence Data; molecular weight; Multigene Family; Mutation; Mycobacterium bovis; NUCLEOTIDE SEQUENCE; Open Reading Frames; OXIDOREDUCTASES; Oxidoreductases - genetics; sequence analysis; Southern blotting; STREPTOMYCES; Streptomyces aureofaciens; Streptomyces aureofaciens - genetics; Streptomyces aureofaciens - metabolism; tchA; tetracycline; tetracycline dehydrogenase; TETRACYCLINES; Streptomycetaceae; Enzyme; Biosynthesis; Identification; coenzyme F420; Dehydrogenase; chlortetracycline biosynthesis; Coenzyme; Tetracycline; Actinomycetales; tetracycline dehydrogenase; Bacteria; Chlortetracycline; Actinomycetes; Oxidoreductases; Molecular cloning; tchA; Streptomyces aureofaciens
• ISSN: 0916-8451; 1347-6947; 1347-6947
• DOI: 10.1271/bbb.68.1345

For chlortetracycline biosynthesis in Streptomyces aureofaciens, the final reduction step is essential to give an antibiotic activity to its intermediate, which is catalyzed by tetracycline dehydrogenase with 7,8-dedimethyl-8-hydroxy-5-deazariboflavin (FO) as a cofactor. We identified and cloned the gene, which is essential for the biosynthesis of 6-demethyltetracycline and participates in the final step of its biosynthesis, from the genomic DNA of the 6-demethyltetracycline producer S. aureofaciens HP77. DNA sequence analysis revealed that the gene (tchA) had an open reading frame of 455 amino acids with an estimated molecular mass of 48.1 kDa. Southern hybridization analysis revealed that the tchA gene was located external to the chlortetracycline biosynthetic gene cluster in the genome. A conserved domain search of protein sequence databases indicated that TchA showed a similarity to FbiB, which is involved in the modification of FO in Mycobacterium bovis.