Biotransformation of phenanthrene and 1-methoxynaphthalene with Streptomyces lividans cells expressing a marine bacterial phenanthrene dioxygenase gene cluster.
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-dihydrophenanthrene by its UV spectral data as well as HPLC property, using the authentic sample for comparison. This biotransformation proceeded very efficiently; 200 microM and 2 mm of phenanthrene were almost completely converted to its cis-diol form in 6 h and 32 h, 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-1,2-dihydro-1,2-naphthalenediol by their EI-MS, 1H- and 13C-NMR spectral data.
Chun
H K
HK
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Japan.
Ohnishi
Y
Y
Misawa
N
N
Shindo
K
K
Hayashi
M
M
Harayama
S
S
Horinouchi
S
S
D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D004274:DNA, Recombinant / Q000096:biosynthesis / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics; D009281:Naphthalenes / Q000378:metabolism*; D010105:Oxygenases / Q000096:biosynthesis / Q000235:genetics*; D010616:Phenanthrenes / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*; D014871:Water Microbiology
D004274:DNA, Recombinant; D009281:Naphthalenes; D010616:Phenanthrenes; C448759:1-methoxynaphthalene; D010105:Oxygenases; C406405:phenanthrene dioxygenase