ID XYL1_CANBO Reviewed; 321 AA.
AC Q8X195;
DT 03-MAY-2011, integrated into UniProtKB/Swiss-Prot.
DT 09-JAN-2007, sequence version 2.
DT 22-FEB-2023, entry version 69.
DE RecName: Full=NADPH-dependent D-xylose reductase;
DE Short=XR;
DE EC=1.1.1.307;
GN Name=XYL1; Synonyms=cbXR;
OS Candida boidinii (Yeast).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC Saccharomycetales; Pichiaceae; Ogataea; Ogataea/Candida clade.
OX NCBI_TaxID=5477;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND SUBSTRATE SPECIFICITY.
RC STRAIN=NRRL Y-17213;
RX PubMed=12721450; DOI=10.1385/abab:106:1-3:265;
RA Kang M.H., Ni H., Jeffries T.W.;
RT "Molecular characterization of a gene for aldose reductase (CbXYL1) from
RT Candida boidinii and its expression in Saccharomyces cerevisiae.";
RL Appl. Biochem. Biotechnol. 106:265-276(2003).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=NRRL Y-17213;
RX PubMed=16838379; DOI=10.1002/bit.21082;
RA Cirino P.C., Chin J.W., Ingram L.O.;
RT "Engineering Escherichia coli for xylitol production from glucose-xylose
RT mixtures.";
RL Biotechnol. Bioeng. 95:1167-1176(2006).
RN [3]
RP FUNCTION.
RX PubMed=18359531; DOI=10.1016/j.jbiotec.2008.02.003;
RA Khankal R., Chin J.W., Cirino P.C.;
RT "Role of xylose transporters in xylitol production from engineered
RT Escherichia coli.";
RL J. Biotechnol. 134:246-252(2008).
RN [4]
RP FUNCTION.
RX PubMed=18698648; DOI=10.1002/bit.22060;
RA Chin J.W., Khankal R., Monroe C.A., Maranas C.D., Cirino P.C.;
RT "Analysis of NADPH supply during xylitol production by engineered
RT Escherichia coli.";
RL Biotechnol. Bioeng. 102:209-220(2009).
RN [5]
RP FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, DOMAIN, AND
RP MUTAGENESIS OF 272-LYS--ASN-274.
RX PubMed=19693930; DOI=10.1002/pro.227;
RA Khoury G.A., Fazelinia H., Chin J.W., Pantazes R.J., Cirino P.C.,
RA Maranas C.D.;
RT "Computational design of Candida boidinii xylose reductase for altered
RT cofactor specificity.";
RL Protein Sci. 18:2125-2138(2009).
CC -!- FUNCTION: Reduces D-xylose into xylitol. Preferentially utilizes NADPH
CC as a cosubstrate. {ECO:0000269|PubMed:12721450,
CC ECO:0000269|PubMed:16838379, ECO:0000269|PubMed:18359531,
CC ECO:0000269|PubMed:18698648, ECO:0000269|PubMed:19693930}.
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=307 uM for NADPH {ECO:0000269|PubMed:19693930};
CC Vmax=152 nmol/min/mg enzyme (in the presence of NADPH)
CC {ECO:0000269|PubMed:19693930};
CC Note=Mutations at positions 272 to 274 modify the cofactor
CC specificity and increase specificity for NADH over NADPH.;
CC -!- PATHWAY: Carbohydrate metabolism; D-xylose degradation.
CC -!- DOMAIN: The Lys-Ser-Asn motif at positions 272 to 274 is important for
CC the cofactor specificity for NADPH over NADH.
CC {ECO:0000269|PubMed:19693930}.
CC -!- SIMILARITY: Belongs to the aldo/keto reductase family. {ECO:0000305}.
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DR EMBL; AF451326; AAL47846.2; -; Genomic_DNA.
DR AlphaFoldDB; Q8X195; -.
DR SMR; Q8X195; -.
DR OrthoDB; 5305445at2759; -.
DR BRENDA; 1.1.1.307; 1100.
DR UniPathway; UPA00810; -.
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; NADP-dependent oxidoreductase domain; 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 PANTHER; PTHR11732; ALDO/KETO REDUCTASE; 1.
DR PANTHER; PTHR11732:SF218; NADPH-DEPENDENT ALDOSE REDUCTASE GRE3; 1.
DR Pfam; PF00248; Aldo_ket_red; 1.
DR PIRSF; PIRSF000097; AKR; 1.
DR PRINTS; PR00069; ALDKETRDTASE.
DR SUPFAM; SSF51430; NAD(P)-linked oxidoreductase; 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 Carbohydrate metabolism; NAD; NADP; Oxidoreductase; Xylose metabolism.
FT CHAIN 1..321
FT /note="NADPH-dependent D-xylose reductase"
FT /id="PRO_0000407987"
FT ACT_SITE 50
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT BINDING 112
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 167..168
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250"
FT BINDING 216..225
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250"
FT BINDING 272..282
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250"
FT SITE 79
FT /note="Lowers pKa of active site Tyr"
FT /evidence="ECO:0000250"
FT MUTAGEN 272..274
FT /note="KSN->EDR: Increases cofactor specificity for NADH
FT over NADPH 240 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->EDR: Increases cofactor specificity for NADH
FT over NADPH more than 1900 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->EDS: Increases cofactor specificity for NADH
FT over NADPH 240 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->EQR: Increases cofactor specificity for NADH
FT over NADPH 6 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->GGD: Increases cofactor specificity for NADH
FT over NADPH 440 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->MAE: Increases cofactor specificity for NADH
FT over NADPH more than 5400 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->MES: Increases cofactor specificity for NADH
FT over NADPH more than 1800 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->MGD: Increases cofactor specificity for NADH
FT over NADPH 890 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->REG: Increases cofactor specificity for NADH
FT over NADPH more than 11200 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->RSE: Increases cofactor specificity for NADH
FT over NADPH 37 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272..274
FT /note="KSN->RTT: Increases cofactor specificity for NADH
FT over NADPH 13 times."
FT /evidence="ECO:0000269|PubMed:19693930"
FT MUTAGEN 272
FT /note="K->R: Increases cofactor specificity for NADH over
FT NADPH 50 times."
SQ SEQUENCE 321 AA; 36470 MW; FCFA8F2A900C86EB CRC64;
MSSPLLTLNN GLKMPQIGFG CWKVDNATCA ETIYEAIKVG YRLFDGAMDY GNEKEVGEGV
NKAIKDGLVK REELFIVSKL WNNFHHPDSV KLAIKKVLSD LNLEYIDLFY MHFPIAQKFV
PIEKKYPPNF YCGDGDKWSF EDVPLLTTWR AMEELVEEGL VKSIGISNFV GALIQDLLRG
CKIRPAVLEI EHHPYLVQPR LIEYAKTEGI HVTAYSSFGP QSFVELDHPK VKDCTTLFKH
ETITSIASAH DVPPAKVLLR WATQRGLAVI PKSNKKERLL GNLKINDFDL TEAELEKIEA
LDIGLRFNDP WTWGYNIPTF I
//
