HSDHB_COLAA
ID HSDHB_COLAA Reviewed; 263 AA.
AC A4ECA9;
DT 17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT 17-APR-2007, sequence version 1.
DT 03-AUG-2022, entry version 66.
DE RecName: Full=7beta-hydroxysteroid dehydrogenase {ECO:0000303|PubMed:21181147, ECO:0000303|PubMed:27006087, ECO:0000303|PubMed:28471355};
DE Short=7beta-HSDH {ECO:0000303|PubMed:21181147};
DE EC=1.1.1.201 {ECO:0000269|PubMed:21181147, ECO:0000269|PubMed:27006087, ECO:0000269|PubMed:6954878};
DE AltName: Full=NADP-dependent 7beta-hydroxysteroid dehydrogenase {ECO:0000303|PubMed:21181147, ECO:0000303|PubMed:6954878};
GN ORFNames=COLAER_02088 {ECO:0000312|EMBL:EBA38809.1};
OS Collinsella aerofaciens (strain ATCC 25986 / DSM 3979 / JCM 10188 / KCTC
OS 3647 / NCTC 11838 / VPI 1003).
OC Bacteria; Actinobacteria; Coriobacteriia; Coriobacteriales;
OC Coriobacteriaceae; Collinsella.
OX NCBI_TaxID=411903;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25986 / DSM 3979 / JCM 10188 / KCTC 3647 / NCTC 11838 / VPI
RC 1003;
RA Sudarsanam P., Ley R., Guruge J., Turnbaugh P.J., Mahowald M., Liep D.,
RA Gordon J.;
RT "Draft genome sequence of Collinsella aerofaciens (ATCC 25986).";
RL Submitted (JAN-2007) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25986 / DSM 3979 / JCM 10188 / KCTC 3647 / NCTC 11838 / VPI
RC 1003;
RA Fulton L., Clifton S., Fulton B., Xu J., Minx P., Mardis E.R., Wilson R.K.;
RL Submitted (APR-2007) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP INDUCTION, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP SUBUNIT.
RC STRAIN=ATCC 25986 / DSM 3979 / JCM 10188 / KCTC 3647 / NCTC 11838 / VPI
RC 1003;
RX PubMed=6954878; DOI=10.1128/aem.43.5.1057-1063.1982;
RA Hirano S., Masuda N.;
RT "Characterization of NADP-dependent 7 beta-hydroxysteroid dehydrogenases
RT from Peptostreptococcus productus and Eubacterium aerofaciens.";
RL Appl. Environ. Microbiol. 43:1057-1063(1982).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=ATCC 25986 / DSM 3979 / JCM 10188 / KCTC 3647 / NCTC 11838 / VPI
RC 1003;
RX PubMed=21181147; DOI=10.1007/s00253-010-3052-y;
RA Liu L., Aigner A., Schmid R.D.;
RT "Identification, cloning, heterologous expression, and characterization of
RT a NADPH-dependent 7beta-hydroxysteroid dehydrogenase from Collinsella
RT aerofaciens.";
RL Appl. Microbiol. Biotechnol. 90:127-135(2011).
RN [5] {ECO:0007744|PDB:5FYD}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP BIOTECHNOLOGY, DELETION MUTANTS, DOMAIN, AND ACTIVE SITE.
RC STRAIN=ATCC 25986 / DSM 3979 / JCM 10188 / KCTC 3647 / NCTC 11838 / VPI
RC 1003;
RX PubMed=27006087; DOI=10.1002/prot.25036;
RA Savino S., Ferrandi E.E., Forneris F., Rovida S., Riva S., Monti D.,
RA Mattevi A.;
RT "Structural and biochemical insights into 7beta-hydroxysteroid
RT dehydrogenase stereoselectivity.";
RL Proteins 84:859-865(2016).
RN [6] {ECO:0007744|PDB:5GT9}
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH NADP, AND SUBUNIT.
RC STRAIN=ATCC 25986 / DSM 3979 / JCM 10188 / KCTC 3647 / NCTC 11838 / VPI
RC 1003;
RX PubMed=28471355; DOI=10.1107/s2053230x17004460;
RA Wang R., Wu J., Jin D.K., Chen Y., Lv Z., Chen Q., Miao Q., Huo X.,
RA Wang F.;
RT "Structure of NADP+-bound 7beta-hydroxysteroid dehydrogenase reveals two
RT cofactor-binding modes.";
RL Acta Crystallogr. F Struct. Biol. Commun. 73:246-252(2017).
CC -!- FUNCTION: 7beta-hydroxysteroid dehydrogenase that catalyzes the
CC reduction of the 7-oxo group of 7-oxo-lithocholate (7-oxo-LCA), to
CC yield ursodeoxycholate (UDCA) (PubMed:21181147, PubMed:6954878,
CC PubMed:27006087). As C.aerofaciens is an intestinal bacterium, this
CC enzyme probably contributes to the formation of UDCA in the human
CC colon. UDCA is regarded as a chemopreventive beneficial secondary bile
CC acid due to its low hydrophobicity; it protects hepatocytes and bile
CC duct epithelial cells against necrosis and apoptosis induced by more
CC hydrophobic secondary bile acids like deoxycholate (DCA) (Probable).
CC This enzyme is also able to catalyze the reverse reaction, i.e. the
CC oxidation of the 7beta-hydroxy group of UDCA to 7-oxo-LCA
CC (PubMed:21181147, PubMed:6954878). To a lesser extent, is also active
CC on the taurine- and glycine-conjugates of ursodeoxycholate. It is
CC specific for NADPH/NADP(+) as the electron acceptor/donor since it is
CC not active with NADH/NAD(+) (PubMed:6954878). In the presence of NADPH,
CC 7beta-HSDH can also reduce dehydrocholate (PubMed:21181147). And is
CC also able to oxidize ursocholate (PubMed:27006087).
CC {ECO:0000269|PubMed:21181147, ECO:0000269|PubMed:27006087,
CC ECO:0000269|PubMed:6954878, ECO:0000305}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 7beta-hydroxysteroid + NADP(+) = a 7-oxosteroid + H(+) +
CC NADPH; Xref=Rhea:RHEA:20233, ChEBI:CHEBI:15378, ChEBI:CHEBI:35349,
CC ChEBI:CHEBI:47789, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC EC=1.1.1.201; Evidence={ECO:0000269|PubMed:21181147,
CC ECO:0000269|PubMed:27006087, ECO:0000269|PubMed:6954878};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:20234;
CC Evidence={ECO:0000269|PubMed:21181147};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:20235;
CC Evidence={ECO:0000269|PubMed:21181147};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7-oxolithocholate + H(+) + NADPH = NADP(+) + ursodeoxycholate;
CC Xref=Rhea:RHEA:47540, ChEBI:CHEBI:15378, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:78604, ChEBI:CHEBI:78605;
CC Evidence={ECO:0000269|PubMed:21181147, ECO:0000269|PubMed:27006087,
CC ECO:0000269|PubMed:6954878};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47541;
CC Evidence={ECO:0000269|PubMed:21181147};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:47542;
CC Evidence={ECO:0000269|PubMed:21181147};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7beta-hydroxy-3,12-dioxo-5beta-cholan-24-oate + NADP(+) =
CC dehydrocholate + H(+) + NADPH; Xref=Rhea:RHEA:53860,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:137881, ChEBI:CHEBI:137882;
CC Evidence={ECO:0000269|PubMed:21181147};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NADP(+) + ursocholate = 3alpha,12alpha-dihydroxy-7-oxo-5beta-
CC cholanate + H(+) + NADPH; Xref=Rhea:RHEA:53856, ChEBI:CHEBI:11893,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:137880; Evidence={ECO:0000269|PubMed:27006087};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.32 uM for NADP(+) {ECO:0000269|PubMed:21181147};
CC KM=4.50 uM for NADPH {ECO:0000269|PubMed:21181147};
CC KM=6.23 uM for ursodeoxycholate {ECO:0000269|PubMed:21181147};
CC KM=5.20 uM for 7-oxolithocholate {ECO:0000269|PubMed:21181147};
CC KM=9.23 uM for dehydrocholate {ECO:0000269|PubMed:21181147};
CC KM=0.108 mM for ursodeoxycholate {ECO:0000269|PubMed:6954878};
CC KM=0.909 mM for glycoursodeoxycholate {ECO:0000269|PubMed:6954878};
CC KM=1.639 mM for tauroursodeoxycholate {ECO:0000269|PubMed:6954878};
CC KM=0.400 mM for NADP(+) {ECO:0000269|PubMed:6954878};
CC Vmax=38.17 umol/min/mg enzyme for the oxidation of ursodeoxycholate
CC {ECO:0000269|PubMed:21181147};
CC Vmax=30.77 umol/min/mg enzyme for the reduction of 7-oxolithocholate
CC {ECO:0000269|PubMed:21181147};
CC Vmax=28.33 umol/min/mg enzyme for the reduction of dehydrocholate
CC {ECO:0000269|PubMed:21181147};
CC Vmax=0.20 umol/min/mg enzyme for the oxidation of ursodeoxycholate
CC {ECO:0000269|PubMed:6954878};
CC Vmax=0.25 umol/min/mg enzyme for the oxidation of
CC glycoursodeoxycholate {ECO:0000269|PubMed:6954878};
CC Vmax=0.24 umol/min/mg enzyme for the oxidation of
CC tauroursodeoxycholate {ECO:0000269|PubMed:6954878};
CC Note=kcat is 1179 min(-1) for the oxidation of ursodeoxycholate. kcat
CC is 951 min(-1) for the reduction of 7-oxolithocholate. kcat is 875
CC min(-1) for the reduction of dehydrocholate.
CC {ECO:0000269|PubMed:21181147};
CC pH dependence:
CC Optimum pH is 9-10 for the oxidation of ursodeoxycholate. Optimum pH
CC is 4-6 for the reduction of 7-oxo-LCA and dehydrocholate.
CC {ECO:0000269|PubMed:21181147};
CC Temperature dependence:
CC Is not very thermostable. The enzyme is completely inactivated at 50
CC degrees Celsius within 5 minutes and at 40 degrees Celsius within 400
CC minutes. {ECO:0000269|PubMed:21181147};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:21181147,
CC ECO:0000269|PubMed:28471355, ECO:0000269|PubMed:6954878}.
CC -!- INDUCTION: Is expressed constitutively and repressed by
CC ursodeoxycholate. {ECO:0000269|PubMed:6954878}.
CC -!- DOMAIN: The 27 C-terminal residues (residues 237-263) are absolutely
CC required for enzymatic activity; a deletion mutant lacking this C-
CC terminal region shows a complete absence of enzymatic activity.
CC {ECO:0000269|PubMed:27006087}.
CC -!- BIOTECHNOLOGY: Could be a useful biocatalyst for the preparative
CC production of ursodeoxycholate, an FDA-approved cholic acid derivate
CC drug used to treat cholesterol gallstones and current sole alternative
CC to surgical intervention. {ECO:0000269|PubMed:27006087}.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
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DR EMBL; AAVN02000010; EBA38809.1; -; Genomic_DNA.
DR RefSeq; WP_006236005.1; NZ_AAVN02000010.1.
DR PDB; 5FYD; X-ray; 1.60 A; A/B=1-263.
DR PDB; 5GT9; X-ray; 1.70 A; A/B=1-263.
DR PDBsum; 5FYD; -.
DR PDBsum; 5GT9; -.
DR AlphaFoldDB; A4ECA9; -.
DR SMR; A4ECA9; -.
DR OrthoDB; 1186478at2; -.
DR BRENDA; 1.1.1.201; 2181.
DR Proteomes; UP000002979; Unassembled WGS sequence.
DR GO; GO:0047022; F:7-beta-hydroxysteroid dehydrogenase (NADP+) activity; IEA:UniProtKB-EC.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0030573; P:bile acid catabolic process; IEA:UniProtKB-KW.
DR GO; GO:0016042; P:lipid catabolic process; IEA:UniProtKB-KW.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR002347; SDR_fam.
DR Pfam; PF00106; adh_short; 1.
DR PRINTS; PR00081; GDHRDH.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Bile acid catabolism; Lipid degradation; Lipid metabolism;
KW NADP; Nucleotide-binding; Oxidoreductase; Steroid metabolism.
FT CHAIN 1..263
FT /note="7beta-hydroxysteroid dehydrogenase"
FT /id="PRO_0000450344"
FT ACT_SITE 156
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:27006087"
FT BINDING 17..21
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:28471355,
FT ECO:0007744|PDB:5GT9"
FT BINDING 40..41
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:28471355,
FT ECO:0007744|PDB:5GT9"
FT BINDING 66..67
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:28471355,
FT ECO:0007744|PDB:5GT9"
FT BINDING 240
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:28471355,
FT ECO:0007744|PDB:5GT9"
FT SITE 143
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:27006087"
FT SITE 160
FT /note="Lowers pKa of active site Tyr"
FT /evidence="ECO:0000250|UniProtKB:P0AET8"
FT HELIX 3..7
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 9..15
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 19..30
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 34..40
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 42..56
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 60..64
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 72..80
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 85..90
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 100..102
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 105..115
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 117..132
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 136..143
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 145..148
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 154..173
FT /evidence="ECO:0007829|PDB:5FYD"
FT TURN 174..176
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 177..186
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 192..195
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 202..208
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 214..224
FT /evidence="ECO:0007829|PDB:5FYD"
FT TURN 225..227
FT /evidence="ECO:0007829|PDB:5FYD"
FT STRAND 229..234
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 235..247
FT /evidence="ECO:0007829|PDB:5FYD"
FT HELIX 250..258
FT /evidence="ECO:0007829|PDB:5FYD"
SQ SEQUENCE 263 AA; 28753 MW; FFF6762088326EAD CRC64;
MNLREKYGEW GLILGATEGV GKAFCEKIAA GGMNVVMVGR REEKLNVLAG EIRETYGVET
KVVRADFSQP GAAETVFAAT EGLDMGFMSY VACLHSFGKI QDTPWEKHEA MINVNVVTFL
KCFHHYMRIF AAQDRGAVIN VSSMTGISSS PWNGQYGAGK AFILKMTEAV ACECEGTGVD
VEVITLGTTL TPSLLSNLPG GPQGEAVMKI ALTPEECVDE AFEKLGKELS VIAGQRNKDS
VHDWKANHTE DEYIRYMGSF YRD
