AFR_ENSAD
ID AFR_ENSAD Reviewed; 333 AA.
AC Q2I8V6;
DT 01-SEP-2009, integrated into UniProtKB/Swiss-Prot.
DT 07-MAR-2006, sequence version 1.
DT 03-AUG-2022, entry version 57.
DE RecName: Full=1,5-anhydro-D-fructose reductase;
DE Short=Anhydrofructose reductase;
DE EC=1.1.1.292;
DE AltName: Full=1,5-anhydro-D-fructose reductase (1,5-anhydro-D-mannitol-forming);
GN Name=afr;
OS Ensifer adhaerens (Sinorhizobium morelense).
OC Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC Rhizobiaceae; Sinorhizobium/Ensifer group; Ensifer.
OX NCBI_TaxID=106592;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-25, CATALYTIC
RP ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, MASS
RP SPECTROMETRY, AND SUBUNIT.
RC STRAIN=S-30.7.5;
RX PubMed=16461673; DOI=10.1128/aem.72.2.1248-1257.2006;
RA Kuehn A., Yu S., Giffhorn F.;
RT "Catabolism of 1,5-anhydro-D-fructose in Sinorhizobium morelense S-30.7.5:
RT discovery, characterization, and overexpression of a new 1,5-anhydro-D-
RT fructose reductase and its application in sugar analysis and rare sugar
RT synthesis.";
RL Appl. Environ. Microbiol. 72:1248-1257(2006).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 2-333 IN COMPLEX WITH NADP, AND
RP MUTAGENESIS OF SER-10; ALA-13; SER-33; LYS-94; ASP-176; HIS-180 AND
RP GLY-206.
RX PubMed=16906761; DOI=10.1021/bi052589q;
RA Dambe T.R., Kuhn A.M., Brossette T., Giffhorn F., Scheidig A.J.;
RT "Crystal structure of NADP(H)-dependent 1,5-anhydro-D-fructose reductase
RT from Sinorhizobium morelense at 2.2 A resolution: construction of a NADH-
RT accepting mutant and its application in rare sugar synthesis.";
RL Biochemistry 45:10030-10042(2006).
CC -!- FUNCTION: Catalyzes the NADPH-specific reduction of 1,5-anhydro-D-
CC fructose to 1,5-anhydro-D-mannitol. Also shows some activity against
CC structurally related compounds such as 3-keto-1,5-anhydro-D-fructose,
CC D-glucosone and D-xylosone. The enzyme cannot use NADH as cosubstrate.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=1,5-anhydro-D-mannitol + NADP(+) = 1,5-anhydro-D-fructose +
CC H(+) + NADPH; Xref=Rhea:RHEA:24208, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16715, ChEBI:CHEBI:49182, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.1.1.292;
CC Evidence={ECO:0000269|PubMed:16461673};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=8.4 mM for 1,5-anhydro-D-fructose {ECO:0000269|PubMed:16461673};
CC KM=11 mM for D-glucosone {ECO:0000269|PubMed:16461673};
CC KM=0.2 mM for NADPH {ECO:0000269|PubMed:16461673};
CC Vmax=500 umol/min/mg enzyme {ECO:0000269|PubMed:16461673};
CC pH dependence:
CC Optimum pH is 6.5 at 30 degrees Celsius.
CC {ECO:0000269|PubMed:16461673};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:16461673,
CC ECO:0000269|PubMed:16906761}.
CC -!- MASS SPECTROMETRY: Mass=35100; Mass_error=130; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:16461673};
CC -!- SIMILARITY: Belongs to the Gfo/Idh/MocA family. {ECO:0000305}.
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DR EMBL; DQ140417; ABA25865.1; -; Genomic_DNA.
DR PDB; 2GLX; X-ray; 2.20 A; A/B/C/D/E/F=2-333.
DR PDBsum; 2GLX; -.
DR AlphaFoldDB; Q2I8V6; -.
DR SMR; Q2I8V6; -.
DR KEGG; ag:ABA25865; -.
DR BioCyc; MetaCyc:MON-16296; -.
DR BRENDA; 1.1.1.292; 8538.
DR SABIO-RK; Q2I8V6; -.
DR EvolutionaryTrace; Q2I8V6; -.
DR GO; GO:0033712; F:1,5-anhydro-D-fructose reductase (1,5-anhydro-D-mannitol-forming) activity; IEA:UniProtKB-EC.
DR GO; GO:0000166; F:nucleotide binding; IEA:InterPro.
DR InterPro; IPR004104; Gfo/Idh/MocA-like_OxRdtase_C.
DR InterPro; IPR000683; Gfo/Idh/MocA-like_OxRdtase_N.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF01408; GFO_IDH_MocA; 1.
DR Pfam; PF02894; GFO_IDH_MocA_C; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; NADP; Oxidoreductase.
FT CHAIN 1..333
FT /note="1,5-anhydro-D-fructose reductase"
FT /id="PRO_0000382696"
FT BINDING 9..12
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 33..34
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 38
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 71..76
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 93..94
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 120
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 162..163
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT BINDING 283
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 10
FT /note="S->G: Almost no effect."
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 13
FT /note="A->G: Can use NAD as cosubstrate as well as NADP."
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 33
FT /note="S->D: No activity."
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 94
FT /note="K->G: Less than 1% remaining activity."
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 176
FT /note="D->A: Less than 1% remaining activity."
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 180
FT /note="H->A: Less than 2% remaining activity."
FT /evidence="ECO:0000269|PubMed:16906761"
FT MUTAGEN 206
FT /note="G->I: No effect."
FT /evidence="ECO:0000269|PubMed:16906761"
FT STRAND 3..8
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 11..15
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 17..23
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 27..32
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 36..45
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 55..59
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 66..69
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 73..75
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 76..85
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 89..92
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 94..96
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 100..113
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 117..119
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 123..125
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 127..137
FT /evidence="ECO:0007829|PDB:2GLX"
FT TURN 138..141
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 142..152
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 157..159
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 162..164
FT /evidence="ECO:0007829|PDB:2GLX"
FT TURN 167..170
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 173..176
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 178..189
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 193..201
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 204..206
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 212..220
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 225..233
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 240..247
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 249..254
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 256..260
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 263..268
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 271..274
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 282..294
FT /evidence="ECO:0007829|PDB:2GLX"
FT STRAND 301..303
FT /evidence="ECO:0007829|PDB:2GLX"
FT HELIX 304..323
FT /evidence="ECO:0007829|PDB:2GLX"
SQ SEQUENCE 333 AA; 35010 MW; A6A088EC1075C917 CRC64;
MNRWGLIGAS TIAREWVIGA IRATGGEVVS MMSTSAERGA AYATENGIGK SVTSVEELVG
DPDVDAVYVS TTNELHREQT LAAIRAGKHV LCEKPLAMTL EDAREMVVAA REAGVVLGTN
HHLRNAAAHR AMRDAIAEGR IGRPIAARVF HAVYLPPHLQ GWRLERPEAG GGVILDITVH
DADTLRFVLN DDPAEAVAIS HSAGMGKEGV EDGVMGGVRF QSGVIAQFHD AFTTKFAETG
FEVHGTEGSL IGRNVMTQKP VGTVTLRNAE GESQLPLDPA NLYETALAAF HSAIEGHGQP
SATGEDGVWS LATGLAVVKA AATGQAAEIE TGL
