RADH_LENKE
ID RADH_LENKE Reviewed; 252 AA.
AC Q6WVP7;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 80.
DE RecName: Full=NADP-dependent (R)-specific alcohol dehydrogenase {ECO:0000303|Ref.1};
DE Short=(R)-specific ADH {ECO:0000303|Ref.1};
DE EC=1.1.1.- {ECO:0000269|Ref.1};
DE AltName: Full=Ketoreductase {ECO:0000303|PubMed:26644568};
DE Short=KRED {ECO:0000303|PubMed:26644568};
GN Name=adh {ECO:0000303|Ref.1};
GN ORFNames=DNL43_05835 {ECO:0000312|EMBL:QGV24812.1},
GN LKE01_04370 {ECO:0000312|EMBL:GEL27617.1};
OS Lentilactobacillus kefiri (Lactobacillus kefiri).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae;
OC Lentilactobacillus.
OX NCBI_TaxID=33962;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY, AND SUBUNIT.
RC STRAIN=ATCC 35411 / DSM 20587 / BCRC 14011 / JCM 5818 / NBRC 15888 / NCIMB
RC 12798 / VKM B-2244 / AK;
RX DOI=10.1080/10242420600893827;
RA Weckbecker A., Hummel W.;
RT "Cloning, expression, and characterization of an (R)-specific alcohol
RT dehydrogenase from Lactobacillus kefir.";
RL Biocatal. Biotransformation 24:380-389(2006).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DH5;
RA Kim D.-H., Seo K.-H.;
RT "Genome sequence of Lactobacillus kefiri DH5 isolated from kefir.";
RL Submitted (JUN-2018) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 35411 / DSM 20587 / BCRC 14011 / JCM 5818 / NBRC 15888 / NCIMB
RC 12798 / VKM B-2244 / AK;
RA Hosoyama A., Uohara A., Ohji S., Ichikawa N.;
RT "Whole genome shotgun sequence of Lactobacillus kefiri NBRC 15888.";
RL Submitted (JUL-2019) to the EMBL/GenBank/DDBJ databases.
RN [4] {ECO:0007744|PDB:4RF2, ECO:0007744|PDB:4RF3, ECO:0007744|PDB:4RF4, ECO:0007744|PDB:4RF5}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF WILD-TYPE AND SEVERAL MUTANTS IN
RP COMPLEX WITH MAGNESIUM AND NADP, ACTIVE SITE, AND ENZYME ENGINEERING.
RX PubMed=26644568; DOI=10.1073/pnas.1507910112;
RA Noey E.L., Tibrewal N., Jimenez-Oses G., Osuna S., Park J., Bond C.M.,
RA Cascio D., Liang J., Zhang X., Huisman G.W., Tang Y., Houk K.N.;
RT "Origins of stereoselectivity in evolved ketoreductases.";
RL Proc. Natl. Acad. Sci. U.S.A. 112:E7065-E7072(2015).
CC -!- FUNCTION: NADP-dependent (R)-specific alcohol dehydrogenase (ADH) with
CC a broad substrate specificity, able to catalyze in vitro the
CC stereoselective reduction of several aliphatic and aromatic ketones as
CC well as beta-keto esters to the corresponding enantiomerically pure
CC alcohols. {ECO:0000269|Ref.1}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a secondary alcohol + NADP(+) = a ketone + H(+) + NADPH;
CC Xref=Rhea:RHEA:19257, ChEBI:CHEBI:15378, ChEBI:CHEBI:17087,
CC ChEBI:CHEBI:35681, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC Evidence={ECO:0000269|Ref.1};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=acetophenone + H(+) + NADPH = (R)-1-phenylethanol + NADP(+);
CC Xref=Rhea:RHEA:68136, ChEBI:CHEBI:15378, ChEBI:CHEBI:27632,
CC ChEBI:CHEBI:45616, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC Evidence={ECO:0000269|Ref.1};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2,5-hexanedione + 2 H(+) + 2 NADPH = (2R,5R)-hexanediol + 2
CC NADP(+); Xref=Rhea:RHEA:68140, ChEBI:CHEBI:15378, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:85014, ChEBI:CHEBI:177024;
CC Evidence={ECO:0000269|Ref.1};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ethyl 3-oxobutanoate + H(+) + NADPH = ethyl (R)-3-
CC hydroxybutanoate + NADP(+); Xref=Rhea:RHEA:68144, ChEBI:CHEBI:4893,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:28707, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; Evidence={ECO:0000269|Ref.1};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-octanone + H(+) + NADPH = (2R)-octan-2-ol + NADP(+);
CC Xref=Rhea:RHEA:68148, ChEBI:CHEBI:15378, ChEBI:CHEBI:37871,
CC ChEBI:CHEBI:57783, ChEBI:CHEBI:58349, ChEBI:CHEBI:87434;
CC Evidence={ECO:0000269|Ref.1};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000269|Ref.1};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.90 mM for acetophenone {ECO:0000269|Ref.1};
CC KM=3.09 mM for (R,S)-1-phenylethanol {ECO:0000269|Ref.1};
CC KM=0.14 mM for NADPH {ECO:0000269|Ref.1};
CC KM=0.03 mM for NADP {ECO:0000269|Ref.1};
CC KM=0.40 mM for NADH {ECO:0000269|Ref.1};
CC KM=0.78 mM for NAD {ECO:0000269|Ref.1};
CC Vmax=400 umol/min/mg enzyme for the NADPH-dependent reduction of
CC acetophenone {ECO:0000269|Ref.1};
CC Vmax=2.9 umol/min/mg enzyme for the NADH-dependent reduction of
CC acetophenone {ECO:0000269|Ref.1};
CC Vmax=15.3 umol/min/mg enzyme for the NADP-dependent oxidation of
CC (R,S)-1-phenylethanol {ECO:0000269|Ref.1};
CC Vmax=0.49 umol/min/mg enzyme for the NAD-dependent oxidation of
CC (R,S)-1-phenylethanol {ECO:0000269|Ref.1};
CC pH dependence:
CC Optimum pH is 7.0 with acetophenone as substrate.
CC {ECO:0000269|Ref.1};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius with acetophenone as
CC substrate. {ECO:0000269|Ref.1};
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|Ref.1}.
CC -!- BIOTECHNOLOGY: Because of its broad substrate specificity and its
CC ability to produce chiral alcohols with high enantiomeric excesses,
CC this protein from L.kefiri is a very interesting enzyme for preparative
CC applications in the synthesis of pharmaceuticals and agrochemicals.
CC {ECO:0000305|Ref.1}.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
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DR EMBL; AY267012; AAP94029.1; -; Genomic_DNA.
DR EMBL; BJVK01000003; GEL27617.1; -; Genomic_DNA.
DR EMBL; CP029971; QGV24812.1; -; Genomic_DNA.
DR RefSeq; WP_054768785.1; NZ_QFFL01000001.1.
DR PDB; 4RF2; X-ray; 2.09 A; A/B=1-252.
DR PDB; 4RF3; X-ray; 1.69 A; A/B=1-252.
DR PDB; 4RF4; X-ray; 2.20 A; A/B=1-252.
DR PDB; 4RF5; X-ray; 1.60 A; A/B=1-252.
DR PDB; 7P36; X-ray; 1.14 A; A=2-252.
DR PDB; 7P7Y; X-ray; 1.25 A; A=2-252.
DR PDBsum; 4RF2; -.
DR PDBsum; 4RF3; -.
DR PDBsum; 4RF4; -.
DR PDBsum; 4RF5; -.
DR PDBsum; 7P36; -.
DR PDBsum; 7P7Y; -.
DR SMR; Q6WVP7; -.
DR STRING; 1423764.FC95_GL000191; -.
DR EnsemblBacteria; PAK84286; PAK84286; B8W85_01105.
DR KEGG; lkf:DNL43_05835; -.
DR Proteomes; UP000321893; Unassembled WGS sequence.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR020904; Sc_DH/Rdtase_CS.
DR InterPro; IPR002347; SDR_fam.
DR PRINTS; PR00081; GDHRDH.
DR PRINTS; PR00080; SDRFAMILY.
DR SUPFAM; SSF51735; SSF51735; 1.
DR PROSITE; PS00061; ADH_SHORT; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Magnesium; Metal-binding; NADP; Nucleotide-binding;
KW Oxidoreductase.
FT CHAIN 1..252
FT /note="NADP-dependent (R)-specific alcohol dehydrogenase"
FT /id="PRO_0000454238"
FT ACT_SITE 156
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000305|PubMed:26644568"
FT BINDING 16..19
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 39..40
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 63..64
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 90
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 156
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 160
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 191..195
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF2"
FT BINDING 252
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:26644568,
FT ECO:0007744|PDB:4RF4"
FT TURN 4..7
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 9..12
FT /evidence="ECO:0007829|PDB:7P36"
FT TURN 13..16
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 18..29
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 33..39
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 41..51
FT /evidence="ECO:0007829|PDB:7P36"
FT TURN 54..56
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 57..61
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 67..81
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 86..89
FT /evidence="ECO:0007829|PDB:7P36"
FT TURN 99..101
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 104..114
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 116..129
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 132..134
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 136..141
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 144..146
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 154..176
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 180..188
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 194..197
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 202..207
FT /evidence="ECO:0007829|PDB:7P36"
FT TURN 209..211
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 220..231
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 233..235
FT /evidence="ECO:0007829|PDB:7P36"
FT STRAND 242..246
FT /evidence="ECO:0007829|PDB:7P36"
FT HELIX 249..251
FT /evidence="ECO:0007829|PDB:7P36"
SQ SEQUENCE 252 AA; 26781 MW; 46A3A79077787670 CRC64;
MTDRLKGKVA IVTGGTLGIG LAIADKFVEE GAKVVITGRH ADVGEKAAKS IGGTDVIRFV
QHDASDEAGW TKLFDTTEEA FGPVTTVVNN AGIAVSKSVE DTTTEEWRKL LSVNLDGVFF
GTRLGIQRMK NKGLGASIIN MSSIEGFVGD PTLGAYNASK GAVRIMSKSA ALDCALKDYD
VRVNTVHPGY IKTPLVDDLE GAEEMMSQRT KTPMGHIGEP NDIAWICVYL ASDESKFATG
AEFVVDGGYT AQ
