title: Identification of 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid in steroid degradation by Comamonas testosteroni TA441 and its conversion to the corresponding 6-en-5-oyl coenzyme A (CoA) involving open reading frame 28 (ORF28)- and ORF30-encoded acyl-CoA dehydrogenases.
abstract: Comamonas testosteroni TA441 degrades steroids via aromatization and meta-cleavage of the A ring, followed by hydrolysis, and produces 9,17-dioxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid as an intermediate compound. Herein, we identify a new intermediate compound, 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid. Open reading frame 28 (ORF28)- and ORF30-encoded acyl coenzyme A (acyl-CoA) dehydrogenase was shown to convert the CoA ester of 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrostan-5-oic acid to the CoA ester of 9α-hydroxy-17-oxo-1,2,3,4,10,19-hexanorandrost-6-en-5-oic acid. A homology search of the deduced amino acid sequences suggested that the ORF30-encoded protein is a member of the acyl-CoA dehydrogenase_fadE6_17_26 family, whereas the deduced amino acid sequence of ORF28 showed no significant similarity to specific acyl-CoA dehydrogenase family proteins. Possible steroid degradation gene clusters similar to the cluster of TA441 appear in bacterial genome analysis data. In these clusters, ORFs similar to ORFs 28 and 30 are often found side by side and ordered in the same manner as ORFs 28 and 30.
authors.affiliation: Environmental Molecular Biology Laboratory, RIKEN, Saitama, Japan masae@postman.riken.go.jp.
authors.forename: Masae
authors.initials: M
authors.lastname: Horinouchi
authors.affiliation: Environmental Molecular Biology Laboratory, RIKEN, Saitama, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Hayashi
authors.affiliation: Molecular Characterization Team, RIKEN, Saitama, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Koshino
authors.affiliation: Molecular Characterization Team, RIKEN, Saitama, Japan.
authors.forename: Michal
authors.initials: M
authors.lastname: Malon
authors.affiliation: Antibiotics Laboratory, RIKEN, Saitama, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hirota
authors.affiliation: Environmental Molecular Biology Laboratory, RIKEN, Saitama, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Kudo
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020604:Comamonas testosteroni / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015394:Molecular Structure; D009154:Mutation; D009654:Norsteroids / Q000737:chemistry / Q000378:metabolism*; D016366:Open Reading Frames; D013256:Steroids / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D009654:Norsteroids; D013256:Steroids
references.citation: J Bacteriol. 1991 Nov;173(22):7219-27
references.pmid: 1657885
references.citation: Nucleic Acids Res. 2000 Nov 1;28(21):4317-31
references.pmid: 11058132
references.citation: Microbiology (Reading). 2001 Dec;147(Pt 12):3367-75
references.pmid: 11739769
references.citation: Nature. 2002 Jan 31;415(6871):497-502
references.pmid: 11823852
references.citation: FEBS Lett. 1985 Aug 19;188(1):85-90
references.pmid: 2991019
references.citation: Nature. 1998 Jun 11;393(6685):537-44
references.pmid: 9634230
references.citation: Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):443-8
references.pmid: 11756688
references.citation: Appl Environ Microbiol. 2005 Sep;71(9):5275-81
references.pmid: 16151114
references.citation: J Steroid Biochem Mol Biol. 2006 Sep;101(1):78-84
references.pmid: 16891113
references.citation: J Steroid Biochem Mol Biol. 2010 Oct;122(4):253-63
references.pmid: 20554032
references.citation: DNA Res. 2002 Dec 31;9(6):225-56
references.pmid: 12597279
references.citation: Nucleic Acids Res. 2013 Jan;41(Database issue):D1096-103
references.pmid: 23087378
references.citation: Eur J Biochem. 1991 Apr 23;197(2):359-65
references.pmid: 2026158
references.citation: Appl Environ Microbiol. 2003 Aug;69(8):4421-30
references.pmid: 12902225
references.citation: J Am Chem Soc. 1965 Mar 20;87:1386-7
references.pmid: 14293760
references.citation: J Steroid Biochem Mol Biol. 2012 Mar;129(1-2):4-14
references.pmid: 21056662
references.citation: Gene. 1987;58(2-3):257-64
references.pmid: 3428616
references.citation: J Bacteriol. 1996 Jun;178(11):3322-30
references.pmid: 8655514
references.citation: J Bacteriol. 2002 Oct;184(19):5479-90
references.pmid: 12218036
references.citation: J Bacteriol. 2004 Mar;186(5):1430-7
references.pmid: 14973025
references.citation: J Steroid Biochem Mol Biol. 2014 Sep;143:306-22
references.pmid: 24810629
references.citation: J Biol Chem. 1998 Nov 20;273(47):30888-96
references.pmid: 9812981
references.citation: Proc Natl Acad Sci U S A. 1987 Dec;84(24):8893-7
references.pmid: 3480517
references.citation: Proc Natl Acad Sci U S A. 2001 Oct 9;98(21):12215-20
references.pmid: 11572948
references.citation: J Biol Chem. 1966 Feb 10;241(3):540-50
references.pmid: 5908120
references.citation: DNA Res. 2002 Dec 31;9(6):189-97
references.pmid: 12597275
references.citation: J Bacteriol. 1993 Oct;175(20):6403-10
references.pmid: 7691794
references.citation: Eur J Biochem. 1996 Nov 1;241(3):744-9
references.pmid: 8944761
references.citation: Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4136-41
references.pmid: 11259647
references.citation: J Biol Chem. 1968 Jun 25;243(12):3238-47
references.pmid: 5656367
references.citation: Biochem J. 1976 Dec 15;160(3):757-68
references.pmid: 1016254
references.citation: Bioinformatics. 2013 Oct 15;29(20):2588-95
references.pmid: 23975762
references.citation: Science. 1999 Nov 19;286(5444):1571-7
references.pmid: 10567266
references.citation: Biochem J. 1983 Mar 15;210(3):829-36
references.pmid: 6870808
references.citation: Biochem J. 1977 Feb 15;162(2):387-97
references.pmid: 849290
references.citation: Nature. 2002 May 9;417(6885):141-7
references.pmid: 12000953
references.citation: Biochem Biophys Res Commun. 2004 Nov 12;324(2):597-604
references.pmid: 15474469
references.citation: J Biol Chem. 1966 Feb 10;241(3):551-9
references.pmid: 5908121
references.citation: J Biol Chem. 1977 Jun 10;252(11):3775-83
references.pmid: 193845
references.citation: J Bacteriol. 2008 Aug;190(16):5545-54
references.pmid: 18539741
references.citation: Biochem J. 1977 Jun 15;164(3):715-26
references.pmid: 883963
references.citation: J Steroid Biochem Mol Biol. 2004 Oct;92(3):143-54
references.pmid: 15555908
references.citation: Biochem J. 1969 Nov;115(2):249-56
references.pmid: 5378382
references.citation: Appl Environ Microbiol. 2003 Apr;69(4):2139-52
references.pmid: 12676694
references.citation: DNA Res. 1999 Apr 30;6(2):83-101, 145-52
references.pmid: 10382966
references.citation: Gene. 1991 Aug 30;105(1):43-9
references.pmid: 1657714