HSDHB_CLOSR
ID HSDHB_CLOSR Reviewed; 261 AA.
AC G9FRD6;
DT 02-DEC-2020, integrated into UniProtKB/Swiss-Prot.
DT 22-FEB-2012, sequence version 1.
DT 03-AUG-2022, entry version 33.
DE RecName: Full=7beta-hydroxysteroid dehydrogenase {ECO:0000303|PubMed:22198717};
DE Short=7beta-HSDH {ECO:0000303|PubMed:22198717};
DE EC=1.1.1.201 {ECO:0000269|PubMed:22198717};
DE AltName: Full=NADP-dependent 7beta-hydroxysteroid dehydrogenase {ECO:0000303|PubMed:22198717};
GN Name=hdhb {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], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND 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).
CC -!- FUNCTION: 7beta-hydroxysteroid dehydrogenase that catalyzes the
CC reduction of the 7-oxo group of 7-oxosteroids, such as 3alpha,12alpha-
CC dihydroxy-7-oxo-5beta-cholanate, 7-oxolithocholate, 7,12-dioxo-
CC lithocholate and dehydrocholate, to the corresponding 7beta-
CC hydroxysteroids. Is also able to catalyze the reverse oxidation
CC reactions. Together with 7alpha-HSDH encoded in the adjacent gene, is
CC likely involved in the epimerization of the hydroxy group at C-7 of
CC primary bile acids through 7-keto bile acid intermediates.
CC {ECO:0000269|PubMed:22198717}.
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:22198717};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:20235;
CC Evidence={ECO:0000305|PubMed:22198717};
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:22198717};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:53858;
CC Evidence={ECO:0000305|PubMed:22198717};
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:22198717};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:47541;
CC Evidence={ECO:0000305|PubMed:22198717};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3alpha,7beta-dihydroxy-12-oxo-5beta-cholan-24-oate + NADP(+) =
CC 7,12-dioxo-lithocholate + H(+) + NADPH; Xref=Rhea:RHEA:53864,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:137789, ChEBI:CHEBI:137886;
CC Evidence={ECO:0000269|PubMed:22198717};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:53866;
CC Evidence={ECO:0000305|PubMed:22198717};
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:22198717};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:53862;
CC Evidence={ECO:0000305|PubMed:22198717};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.59 mM for ursocholate {ECO:0000269|PubMed:22198717};
CC KM=2.30 mM for 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholanate
CC {ECO:0000269|PubMed:22198717};
CC KM=3.06 mM for ursodeoxycholate {ECO:0000269|PubMed:22198717};
CC KM=2.65 mM for 7-oxolithocholate {ECO:0000269|PubMed:22198717};
CC KM=1.50 mM for 7,12-dioxo-lithocholate {ECO:0000269|PubMed:22198717};
CC KM=1.58 mM for dehydrocholate {ECO:0000269|PubMed:22198717};
CC Note=kcat is 407000 sec(-1) for the oxidation of ursocholate. kcat is
CC 203000 sec(-1) for the reduction of 3alpha,12alpha-dihydroxy-7-oxo-
CC 5beta-cholanate. kcat is 430000 sec(-1) for the oxidation of
CC ursodeoxycholate. kcat is 639000 sec(-1) for the reduction of 7-
CC oxolithocholate. kcat is 413000 sec(-1) for the reduction of 7,12-
CC dioxo-lithocholate. kcat is 146000 sec(-1) for the reduction of
CC dehydrocholate. {ECO:0000269|PubMed:22198717};
CC pH dependence:
CC Shows a maximum of activity at pH 8.5 in the oxidation of
CC ursocholate, and between pH 7.0 and 8.0 when tested in the reduction
CC reaction of 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholanate, with a
CC gradual drop on the alkaline side and a sharp drop on the acidic
CC side. {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: Homodimer. {ECO:0000269|PubMed:22198717}.
CC -!- INDUCTION: Induced 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; AET80684.1; -; Genomic_DNA.
DR AlphaFoldDB; G9FRD6; -.
DR SMR; G9FRD6; -.
DR SwissLipids; SLP:000001739; -.
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 Bile acid catabolism; Lipid degradation; Lipid metabolism; NADP;
KW Nucleotide-binding; Oxidoreductase; Steroid metabolism.
FT CHAIN 1..261
FT /note="7beta-hydroxysteroid dehydrogenase"
FT /id="PRO_0000451450"
FT ACT_SITE 156
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:A4ECA9"
FT BINDING 17..21
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:A4ECA9"
FT BINDING 40..41
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:A4ECA9"
FT BINDING 66..67
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:A4ECA9"
FT SITE 143
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:A4ECA9"
FT SITE 160
FT /note="Lowers pKa of active site Tyr"
FT /evidence="ECO:0000250|UniProtKB:P0AET8"
SQ SEQUENCE 261 AA; 29383 MW; 148C510A89EC17AC CRC64;
MNFREKYGQW GIVLGATEGI GKASAFELAK RGMDVILVGR RKEALEELAK AIHEETGKEI
RVLPQDLSEY DAAERLIEAT KDLDMGVIEY VACLHAMGQY NKVDYAKYEQ MYRVNIRTFS
KLLHHYIGEF KERDRGAFIT IGSLSGWTSL PFCAEYAAEK AYMMTVTEGV AYECANTNVD
VMLLSAGSTI TPTWLKNKPS DPKAVAAAMY PEDVIKDGFE QLGKKFTYLA GELNREKMKE
NNAMDRNDLI AKLGKMFDHM A
