HDHA_CLOSR
ID HDHA_CLOSR Reviewed; 262 AA.
AC G9FRD7;
DT 02-DEC-2020, integrated into UniProtKB/Swiss-Prot.
DT 22-FEB-2012, sequence version 1.
DT 03-AUG-2022, entry version 37.
DE RecName: Full=7alpha-hydroxysteroid dehydrogenase {ECO:0000303|PubMed:22198717, ECO:0000303|PubMed:26961171};
DE Short=7alpha-HSDH {ECO:0000303|PubMed:22198717, ECO:0000303|PubMed:26961171};
DE EC=1.1.1.- {ECO:0000269|PubMed:22198717, ECO:0000269|PubMed:24810359};
DE AltName: Full=NADP-dependent 7alpha-hydroxysteroid dehydrogenase {ECO:0000303|PubMed:22198717};
GN Name=hdha {ECO:0000303|PubMed:22198717};
OS Clostridium sardiniense (Clostridium absonum).
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae;
OC Clostridium.
OX NCBI_TaxID=29369;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-27, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND
RP BIOTECHNOLOGY.
RC STRAIN=ATCC 27555 / DSM 599 / CIP 104302 / JCM 1381 / NCTC 10984 / HA 7103;
RX PubMed=22198717; DOI=10.1007/s00253-011-3798-x;
RA Ferrandi E.E., Bertolesi G.M., Polentini F., Negri A., Riva S., Monti D.;
RT "In search of sustainable chemical processes: cloning, recombinant
RT expression, and functional characterization of the 7alpha- and 7beta-
RT hydroxysteroid dehydrogenases from Clostridium absonum.";
RL Appl. Microbiol. Biotechnol. 95:1221-1223(2012).
RN [2]
RP INDUCTION BY BILE ACIDS.
RX PubMed=6945134; DOI=10.1016/0005-2760(81)90011-4;
RA MacDonald I.A., Roach P.D.;
RT "Bile induction of 7 alpha- and 7 beta-hydroxysteroid dehydrogenases in
RT Clostridium absonum.";
RL Biochim. Biophys. Acta 665:262-269(1981).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND MUTAGENESIS OF ARG-38 AND 261-PRO-ARG-262.
RX PubMed=24810359; DOI=10.2174/0929866521666140507160050;
RA Lou D., Wang B., Tan J., Zhu L.;
RT "Carboxyl-terminal and Arg38 are essential for activity of the 7alpha-
RT hydroxysteroid dehydrogenase from Clostridium absonum.";
RL Protein Pept. Lett. 21:894-900(2014).
RN [4] {ECO:0007744|PDB:5EPO}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEX WITH NADP AND
RP TAUROCHENODEOXYCHOLATE, AND SUBUNIT.
RX PubMed=26961171; DOI=10.1038/srep22885;
RA Lou D., Wang B., Tan J., Zhu L., Cen X., Ji Q., Wang Y.;
RT "The three-dimensional structure of Clostridium absonum 7alpha-
RT hydroxysteroid dehydrogenase: new insights into the conserved arginines for
RT NADP(H) recognition.";
RL Sci. Rep. 6:22885-22885(2016).
CC -!- FUNCTION: 7alpha-hydroxysteroid dehydrogenase that catalyzes the
CC NADP(+)-dependent oxidation of the 7alpha-hydroxy group of 7alpha-
CC hydroxysteroids, such as cholate, chenodeoxycholate,
CC glycochenodeoxycholate and taurochenodeoxycholate, to the corresponding
CC 7-oxosteroids (PubMed:22198717, PubMed:24810359). Is also able to
CC catalyze the reverse reduction reactions (PubMed:22198717). Together
CC with 7beta-HSDH encoded in the adjacent gene, is likely involved in the
CC epimerization of the hydroxy group at C-7 of primary bile acids through
CC 7-keto bile acid intermediates (PubMed:22198717).
CC {ECO:0000269|PubMed:22198717, ECO:0000269|PubMed:24810359}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cholate + NADP(+) = 3alpha,12alpha-dihydroxy-7-oxo-5beta-
CC cholanate + H(+) + NADPH; Xref=Rhea:RHEA:48508, ChEBI:CHEBI:11893,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29747, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; Evidence={ECO:0000269|PubMed:22198717};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:48509;
CC Evidence={ECO:0000305|PubMed:22198717};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=chenodeoxycholate + NADP(+) = 7-oxolithocholate + H(+) +
CC NADPH; Xref=Rhea:RHEA:53820, ChEBI:CHEBI:15378, ChEBI:CHEBI:36234,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:78605;
CC Evidence={ECO:0000269|PubMed:22198717, ECO:0000269|PubMed:24810359};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:53821;
CC Evidence={ECO:0000305|PubMed:22198717};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate + NADP(+) =
CC 7,12-dioxo-lithocholate + H(+) + NADPH; Xref=Rhea:RHEA:53840,
CC ChEBI:CHEBI:11901, ChEBI:CHEBI:15378, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:137789;
CC Evidence={ECO:0000269|PubMed:22198717};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:53841;
CC Evidence={ECO:0000305|PubMed:22198717};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7alpha-hydroxy-3,12-dioxo-5beta-cholanate + NADP(+) =
CC dehydrocholate + H(+) + NADPH; Xref=Rhea:RHEA:65024,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:137881, ChEBI:CHEBI:156270;
CC Evidence={ECO:0000269|PubMed:22198717};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:65025;
CC Evidence={ECO:0000305|PubMed:22198717};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=glycochenodeoxycholate + NADP(+) = 7-oxoglycolithocholate +
CC H(+) + NADPH; Xref=Rhea:RHEA:65056, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:36252, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:137818; Evidence={ECO:0000269|PubMed:24810359};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:65057;
CC Evidence={ECO:0000305|PubMed:24810359};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NADP(+) + taurochenodeoxycholate = 7-oxotaurolithocholate +
CC H(+) + NADPH; Xref=Rhea:RHEA:65060, ChEBI:CHEBI:9407,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:137724; Evidence={ECO:0000269|PubMed:24810359};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:65061;
CC Evidence={ECO:0000305|PubMed:24810359};
CC -!- ACTIVITY REGULATION: Activated by metal ions such as Mg(2+), Na(+) and
CC K(+). {ECO:0000269|PubMed:24810359}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.06 mM for cholate {ECO:0000269|PubMed:22198717};
CC KM=0.96 mM for 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholanate
CC {ECO:0000269|PubMed:22198717};
CC KM=0.09 mM for chenodeoxycholate {ECO:0000269|PubMed:22198717};
CC KM=0.13 mM for 7-oxolithocholate {ECO:0000269|PubMed:22198717};
CC KM=5.70 mM for 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate
CC {ECO:0000269|PubMed:22198717};
CC KM=1.14 mM for 7,12-dioxo-lithocholate {ECO:0000269|PubMed:22198717};
CC KM=3.50 mM for dehydrocholate {ECO:0000269|PubMed:22198717};
CC KM=0.013 mM for chenodeoxycholate {ECO:0000269|PubMed:24810359};
CC KM=0.012 mM for glycochenodeoxycholate {ECO:0000269|PubMed:24810359};
CC KM=0.016 mM for taurochenodeoxycholate {ECO:0000269|PubMed:24810359};
CC Note=kcat is 343000 sec(-1) for the oxidation of cholate. kcat is
CC 13600 sec(-1) for the reduction of 3alpha,12alpha-dihydroxy-7-oxo-
CC 5beta-cholanate. kcat is 9390 sec(-1) for the oxidation of
CC chenodeoxycholate. kcat is 977 sec(-1) for the reduction of 7-
CC oxolithocholate. kcat is 222000 sec(-1) for the oxidation of
CC 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate. kcat is 30600 sec(-1)
CC for the reduction of 7,12-dioxo-lithocholate. kcat is 22600 sec(-1)
CC for the reduction of dehydrocholate (PubMed:22198717). kcat is 21.2
CC sec(-1) for the oxidation of chenodeoxycholate. kcat is 19.6 sec(-1)
CC for the oxidation of glycochenodeoxycholate. kcat is 11.5 sec(-1) for
CC the oxidation of taurochenodeoxycholate (PubMed:24810359).
CC {ECO:0000269|PubMed:22198717, ECO:0000269|PubMed:24810359};
CC pH dependence:
CC Shows a maximum of activity between pH 7.5 and 8.0 both in the
CC oxidation reaction of cholate and in the reduction reaction of
CC 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholanate.
CC {ECO:0000269|PubMed:22198717};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius. No activity is detected
CC above 60 degrees Celsius. {ECO:0000269|PubMed:22198717};
CC -!- SUBUNIT: Homotetramer (PubMed:22198717, PubMed:26961171). A dynamic
CC equilibrium between dimers and tetramers seems to exist
CC (PubMed:26961171). {ECO:0000269|PubMed:22198717,
CC ECO:0000269|PubMed:26961171}.
CC -!- INDUCTION: Up-regulated by bile acids, when grown in the presence of
CC deoxycholate or chenodeoxycholate. {ECO:0000269|PubMed:6945134}.
CC -!- BIOTECHNOLOGY: Could be a promising candidate for further applications
CC in the epimerization reaction of bile acids at the C-7 position and
CC thus may be used as a biocatalyst for the synthesis of bile acids
CC derivatives of pharmacological interest. {ECO:0000305|PubMed:22198717}.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
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DR EMBL; JN191345; AET80685.1; -; Genomic_DNA.
DR PDB; 5EPO; X-ray; 2.00 A; A/B/C/D=1-262.
DR PDBsum; 5EPO; -.
DR AlphaFoldDB; G9FRD7; -.
DR SMR; G9FRD7; -.
DR SwissLipids; SLP:000001737; -.
DR BRENDA; 1.1.1.159; 1450.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; 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 PRINTS; PR00081; GDHRDH.
DR PRINTS; PR00080; SDRFAMILY.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Bile acid catabolism; Direct protein sequencing;
KW Lipid degradation; Lipid metabolism; NADP; Nucleotide-binding;
KW Oxidoreductase; Steroid metabolism.
FT CHAIN 1..262
FT /note="7alpha-hydroxysteroid dehydrogenase"
FT /id="PRO_0000451449"
FT ACT_SITE 158
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:P0AET8"
FT BINDING 13..18
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 38
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 63..64
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 90
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 145
FT /ligand="taurochenodeoxycholate"
FT /ligand_id="ChEBI:CHEBI:9407"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 158
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 158
FT /ligand="taurochenodeoxycholate"
FT /ligand_id="ChEBI:CHEBI:9407"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 162
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT BINDING 191..195
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26961171,
FT ECO:0007744|PDB:5EPO"
FT SITE 162
FT /note="Lowers pKa of active site Tyr"
FT /evidence="ECO:0000250|UniProtKB:P0AET8"
FT MUTAGEN 38
FT /note="R->D: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:24810359"
FT MUTAGEN 261..262
FT /note="Missing: 5-fold reduction in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:24810359"
FT TURN 3..6
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 8..13
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 17..28
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 32..38
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 40..52
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 56..61
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 68..81
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 86..89
FT /evidence="ECO:0007829|PDB:5EPO"
FT TURN 96..98
FT /evidence="ECO:0007829|PDB:5EPO"
FT TURN 102..104
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 107..134
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 138..143
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 146..148
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 156..176
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 177..179
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 181..188
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 194..199
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 202..210
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 220..231
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 233..235
FT /evidence="ECO:0007829|PDB:5EPO"
FT STRAND 242..246
FT /evidence="ECO:0007829|PDB:5EPO"
FT TURN 247..250
FT /evidence="ECO:0007829|PDB:5EPO"
FT HELIX 256..259
FT /evidence="ECO:0007829|PDB:5EPO"
SQ SEQUENCE 262 AA; 28289 MW; 10D0D76D2A88EFB6 CRC64;
MKRLEGKVAI VTSSTRGIGR ASAEALAKEG ALVYLAARSE ELANEVIADI KKQGGVAKFV
YFNAREEETY TSMVEKVAEA EGRIDILVNN YGGTNVNLDK NLTAGDTDEF FRILKDNVQS
VYLPAKAAIP HMEKVGGGSI VNISTIGSVV PDISRIAYCV SKSAINSLTQ NIALQYARKN
IRCNAVLPGL IGTRAALENM TDEFRDSFLG HVPLNRVGRP EDIANAVLYY ASDDSGYVTG
MIHEVAGGFA LGTPQYSEYC PR
