XYL1_CANTE
ID XYL1_CANTE Reviewed; 322 AA.
AC O74237;
DT 11-OCT-2004, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1998, sequence version 1.
DT 03-AUG-2022, entry version 87.
DE RecName: Full=NAD(P)H-dependent D-xylose reductase;
DE Short=XR;
DE EC=1.1.1.307;
GN Name=XYL1; Synonyms=XYLR;
OS Candida tenuis (Yeast) (Yamadazyma tenuis).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC Saccharomycetales; Debaryomycetaceae; Yamadazyma.
OX NCBI_TaxID=2315449;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=CBS 4435;
RX PubMed=10661866; DOI=10.1515/bc.1999.179;
RA Hacker B., Habenicht A., Kiess M., Mattes R.;
RT "Xylose utilisation: cloning and characterisation of the xylose reductase
RT from Candida tenuis.";
RL Biol. Chem. 380:1395-1403(1999).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS), AND SUBUNIT.
RX PubMed=12102621; DOI=10.1021/bi025786n;
RA Kavanagh K.L., Klimacek M., Nidetzky B., Wilson D.K.;
RT "The structure of apo and holo forms of xylose reductase, a dimeric aldo-
RT keto reductase from Candida tenuis.";
RL Biochemistry 41:8785-8795(2002).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) IN COMPLEX WITH NAD, AND SUBUNIT.
RX PubMed=12733986; DOI=10.1042/bj20030286;
RA Kavanagh K.L., Klimacek M., Nidetzky B., Wilson D.K.;
RT "Structure of xylose reductase bound to NAD+ and the basis for single and
RT dual co-substrate specificity in family 2 aldo-keto reductases.";
RL Biochem. J. 373:319-326(2003).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT ARG-274/ASP-276 IN COMPLEX
RP WITH NAD, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF LYS-274;
RP SER-275; ASN-276 AND ARG-280.
RX PubMed=15320875; DOI=10.1042/bj20040363;
RA Petschacher B., Leitgeb S., Kavanagh K.L., Wilson D.K., Nidetzky B.;
RT "The coenzyme specificity of Candida tenuis xylose reductase (AKR2B5)
RT explored by site-directed mutagenesis and X-ray crystallography.";
RL Biochem. J. 385:75-83(2005).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS) OF MUTANT ARG-274 IN COMPLEXES WITH
RP NAD AND NADP.
RX PubMed=15670843; DOI=10.1016/j.febslet.2004.12.063;
RA Leitgeb S., Petschacher B., Wilson D.K., Nidetzky B.;
RT "Fine tuning of coenzyme specificity in family 2 aldo-keto reductases
RT revealed by crystal structures of the Lys-274-->Arg mutant of Candida
RT tenuis xylose reductase (AKR2B5) bound to NAD+ and NADP+.";
RL FEBS Lett. 579:763-767(2005).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 2-322 OF MUTANT ASP-309 IN COMPLEX
RP WITH NAD, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF TRP-24; ASP-51
RP AND ASN-310.
RX PubMed=16336198; DOI=10.1042/bj20050831;
RA Kratzer R., Leitgeb S., Wilson D.K., Nidetzky B.;
RT "Probing the substrate binding site of Candida tenuis xylose reductase
RT (AKR2B5) with site-directed mutagenesis.";
RL Biochem. J. 393:51-58(2006).
CC -!- FUNCTION: Reduces D-xylose into xylitol. Has a preference for NADPH,
CC but can also utilize NADH as cosubstrate.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NAD(+) + xylitol = D-xylose + H(+) + NADH;
CC Xref=Rhea:RHEA:27441, ChEBI:CHEBI:15378, ChEBI:CHEBI:17151,
CC ChEBI:CHEBI:53455, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945;
CC EC=1.1.1.307;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=NADP(+) + xylitol = D-xylose + H(+) + NADPH;
CC Xref=Rhea:RHEA:27445, ChEBI:CHEBI:15378, ChEBI:CHEBI:17151,
CC ChEBI:CHEBI:53455, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC EC=1.1.1.307;
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=142 mM for xylose {ECO:0000269|PubMed:15320875,
CC ECO:0000269|PubMed:16336198};
CC KM=38 uM for NADH {ECO:0000269|PubMed:15320875,
CC ECO:0000269|PubMed:16336198};
CC KM=3 uM for NADPH {ECO:0000269|PubMed:15320875,
CC ECO:0000269|PubMed:16336198};
CC -!- PATHWAY: Carbohydrate metabolism; D-xylose degradation.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:12102621,
CC ECO:0000269|PubMed:12733986, ECO:0000269|PubMed:15320875,
CC ECO:0000269|PubMed:16336198}.
CC -!- SIMILARITY: Belongs to the aldo/keto reductase family. {ECO:0000305}.
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DR EMBL; AF074484; AAC25601.1; -; Genomic_DNA.
DR PDB; 1JEZ; X-ray; 2.20 A; A/B=1-322.
DR PDB; 1K8C; X-ray; 2.10 A; A/B/C/D=1-322.
DR PDB; 1MI3; X-ray; 1.80 A; A/B/C/D=1-322.
DR PDB; 1R38; X-ray; 2.20 A; A/B/C/D=1-322.
DR PDB; 1SM9; X-ray; 2.20 A; A/B/C/D=1-322.
DR PDB; 1YE4; X-ray; 2.40 A; A/B/C/D=1-322.
DR PDB; 1YE6; X-ray; 2.30 A; A/B/C/D=1-322.
DR PDB; 1Z9A; X-ray; 2.40 A; A/B/C/D=2-322.
DR PDBsum; 1JEZ; -.
DR PDBsum; 1K8C; -.
DR PDBsum; 1MI3; -.
DR PDBsum; 1R38; -.
DR PDBsum; 1SM9; -.
DR PDBsum; 1YE4; -.
DR PDBsum; 1YE6; -.
DR PDBsum; 1Z9A; -.
DR AlphaFoldDB; O74237; -.
DR SMR; O74237; -.
DR BRENDA; 1.1.1.307; 1144.
DR SABIO-RK; O74237; -.
DR UniPathway; UPA00810; -.
DR EvolutionaryTrace; O74237; -.
DR GO; GO:0032866; F:D-xylose:NADP reductase activity; IEA:InterPro.
DR GO; GO:0042843; P:D-xylose catabolic process; IEA:UniProtKB-UniPathway.
DR CDD; cd19113; AKR_AKR2B1-10; 1.
DR Gene3D; 3.20.20.100; -; 1.
DR InterPro; IPR020471; AKR.
DR InterPro; IPR044486; AKR2B1.
DR InterPro; IPR018170; Aldo/ket_reductase_CS.
DR InterPro; IPR023210; NADP_OxRdtase_dom.
DR InterPro; IPR036812; NADP_OxRdtase_dom_sf.
DR Pfam; PF00248; Aldo_ket_red; 1.
DR PIRSF; PIRSF000097; AKR; 1.
DR PRINTS; PR00069; ALDKETRDTASE.
DR SUPFAM; SSF51430; SSF51430; 1.
DR PROSITE; PS00798; ALDOKETO_REDUCTASE_1; 1.
DR PROSITE; PS00062; ALDOKETO_REDUCTASE_2; 1.
DR PROSITE; PS00063; ALDOKETO_REDUCTASE_3; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; NAD; NADP; Oxidoreductase;
KW Xylose metabolism.
FT CHAIN 1..322
FT /note="NAD(P)H-dependent D-xylose reductase"
FT /id="PRO_0000124661"
FT ACT_SITE 52
FT /note="Proton donor"
FT BINDING 114
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 169..170
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:12733986,
FT ECO:0000269|PubMed:15320875, ECO:0000269|PubMed:16336198"
FT BINDING 218..227
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:12733986,
FT ECO:0000269|PubMed:15320875, ECO:0000269|PubMed:16336198"
FT BINDING 274..284
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:12733986,
FT ECO:0000269|PubMed:15320875, ECO:0000269|PubMed:16336198"
FT SITE 81
FT /note="Lowers pKa of active site Tyr"
FT /evidence="ECO:0000250"
FT MUTAGEN 24
FT /note="W->F,Y: Strongly reduced affinity for xylose.
FT Reduces NADH-dependent enzyme activity by over 96%."
FT /evidence="ECO:0000269|PubMed:16336198"
FT MUTAGEN 51
FT /note="D->A: Slightly reduced enzyme activity."
FT /evidence="ECO:0000269|PubMed:16336198"
FT MUTAGEN 274
FT /note="K->E: Reduces enzyme activity about 1000-fold."
FT /evidence="ECO:0000269|PubMed:15320875"
FT MUTAGEN 274
FT /note="K->G,M: Reduces affinity for NAD and NADP."
FT /evidence="ECO:0000269|PubMed:15320875"
FT MUTAGEN 274
FT /note="K->R: Increases affinity for NAD. Strongly reduced
FT enzyme activity with NADP; when associated with D-276."
FT /evidence="ECO:0000269|PubMed:15320875"
FT MUTAGEN 275
FT /note="S->A: Decreases affinity for NAD and NADP."
FT /evidence="ECO:0000269|PubMed:15320875"
FT MUTAGEN 276
FT /note="N->D: Increases affinity for NAD. Decreases affinity
FT for NADP. Strongly reduced enzyme activity with NADP; when
FT associated with R-274."
FT /evidence="ECO:0000269|PubMed:15320875"
FT MUTAGEN 280
FT /note="R->H: Increases affinity for NAD."
FT /evidence="ECO:0000269|PubMed:15320875"
FT MUTAGEN 310
FT /note="N->A,D: Strongly decreased affinity for xylose."
FT /evidence="ECO:0000269|PubMed:16336198"
FT STRAND 7..9
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 15..19
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 28..40
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 45..47
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 50..52
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 55..67
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 73..75
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 77..82
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 84..86
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 89..103
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 108..113
FT /evidence="ECO:0007829|PDB:1MI3"
FT TURN 124..126
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 147..159
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 162..170
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 173..182
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 189..193
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 201..209
FT /evidence="ECO:0007829|PDB:1MI3"
FT STRAND 213..217
FT /evidence="ECO:0007829|PDB:1MI3"
FT TURN 219..222
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 223..226
FT /evidence="ECO:0007829|PDB:1MI3"
FT TURN 227..229
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 231..234
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 239..241
FT /evidence="ECO:0007829|PDB:1K8C"
FT HELIX 243..252
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 256..265
FT /evidence="ECO:0007829|PDB:1MI3"
FT TURN 266..268
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 278..283
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 294..301
FT /evidence="ECO:0007829|PDB:1MI3"
FT HELIX 313..316
FT /evidence="ECO:0007829|PDB:1MI3"
SQ SEQUENCE 322 AA; 36021 MW; 4C74A8FBC9357690 CRC64;
MSASIPDIKL SSGHLMPSIG FGCWKLANAT AGEQVYQAIK AGYRLFDGAE DYGNEKEVGD
GVKRAIDEGL VKREEIFLTS KLWNNYHDPK NVETALNKTL ADLKVDYVDL FLIHFPIAFK
FVPIEEKYPP GFYCGDGNNF VYEDVPILET WKALEKLVAA GKIKSIGVSN FPGALLLDLL
RGATIKPAVL QVEHHPYLQQ PKLIEFAQKA GVTITAYSSF GPQSFVEMNQ GRALNTPTLF
AHDTIKAIAA KYNKTPAEVL LRWAAQRGIA VIPKSNLPER LVQNRSFNTF DLTKEDFEEI
AKLDIGLRFN DPWDWDNIPI FV
