Biotransformation of phenanthrene and 1-methoxynaphthalene with Streptomyces lividans cells expressing a marine bacterial phenanthrene dioxygenase gene cluster

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2001-08, Vol.65 (8), p.1774-1781
• Main Authors: Chun, H.K. (Tokyo Univ. (Japan)), Ohnishi, Y, Misawa, N, Shindo, K, Hayashi, M, Harayama, S, Horinouchi, S
• Format: Article
• Language: English
• Subjects: 1-methoxynaphthalene; 8-methoxy-1,2-dihydro-1,2-naphthalenediol; 8-methoxy-2-naphthol; Actinomycetales - enzymology; Actinomycetales - genetics; AQUATIC ORGANISMS; arene dioxygenase; AROMATIC COMPOUNDS; BIOCHEMISTRY; Bioconversions. Hemisynthesis; Biodegradation of pollutants; Biological and medical sciences; Biotechnology; biotransformation; Chromatography, High Pressure Liquid; DNA, Recombinant - biosynthesis; DNA, Recombinant - chemistry; Environment and pollution; Escherichia coli - genetics; Escherichia coli - metabolism; ferredoxin; Fundamental and applied biological sciences. Psychology; GENE EXPRESSION; GENES; Genes, Bacterial - genetics; Industrial applications and implications. Economical aspects; Methods. Procedures. Technologies; Naphthalenes - metabolism; Nocardioides; OXIDOREDUCTASES; Oxygenases - biosynthesis; Oxygenases - genetics; phenanthrene; Phenanthrenes - metabolism; Plasmids - genetics; STREPTOMYCES; Streptomyces - genetics; Streptomyces - metabolism; Streptomyces lividans; Water Microbiology
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.65.1774

The phdABCD gene cluster in a marine bacterium Nocardioides sp. strain KP7 codes for the multicomponent enzyme phenanthrene dioxygenase. phdA encoding an iron-sulfur protein large subunit alpha, phdB encoding its small subunit beta, phdC encoding ferredoxin, and phdD encoding ferredoxin reductase, were replaced in such a way that the termination codons of the preceding open reading frames were overlapped with the initiation codons of the following genes. This manipulated phdABCD gene cluster was positioned downstream of the thiostrepton-inducible promoter PtipA in a high-copy-number vector pIJ6021, and introduced into the gram-positive, soil-inhabiting, filamentous bacterium Streptomyces lividans. The recombinant S.lividans cells converted phenanthrene into a cis-diol form, which was determined to be cis-3,4-dihydroxy-3, 4-dihydrophenan-threne by its UV spectral data as well as HPLC property, using the authentic sample for comparison. This biotransformation proceeded very efficiently; 200 micro M and 2 mM of phenanthrene were almost completely converted to its cis-diol form in 6h and 32h, respectively. In addition, the S.lividans cells carrying the phdABCD gene cluster were found to transform 1-methoxynaphthalene to two products, which were identified to be 8-methoxy-2-naphthol in addition to 8-methoxy-l, 2-dihydro-l, 2-naphthalenediol by their EI-MS, 1H- and 13C-NMR spectral data.