The genes encoding the hydroxylase of 3-hydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione in steroid degradation in Comamonas testosteroni TA441.
Steroid degradation genes of Comamonas testosteroni TA441 are encoded in at least two gene clusters: one containing the meta-cleavage enzyme gene tesB; and another consisting of ORF18, 17, tesI, H, ORF11, 12, and tesDEFG. TesH and I are, respectively, the Delta(1)- and Delta(4)(5alpha)-dehydrogenase of the 3-ketosteroid, TesD is the hydrolase for the product of meta-cleavage reaction, and TesEFG degrade one of the product of TesD. In this report, we describe the identification of the function of ORF11 (tesA2) and 12 (tesA1). The TesA1- and TesA2-disrupted mutant accumulated two characteristic intermediate compounds, which were identified as 3-hydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione (3-HSA) and its hydroxylated derivative, 3,17-dihydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione by MS and NMR analysis. A complementation experiment using a broad-host range plasmid showed that both TesA1 and A2 are necessary for hydroxylation of 3-HSA to 3,4-dihydroxy-9,10-secoandrosta-1,3,5(10)-triene-9,17-dione (3,4-DHSA).
Horinouchi
Masae
M
RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan. masae@riken.jp
Hayashi
Toshiaki
T
Kudo
Toshiaki
T
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D002874:Chromosome Mapping; D020604:Comamonas testosteroni / Q000201:enzymology* / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013250:Steroid Hydroxylases / Q000235:genetics* / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism
D013739:Testosterone; D013250:Steroid Hydroxylases