FDH_CANBO
ID FDH_CANBO Reviewed; 364 AA.
AC O13437; O93968; Q00498; Q1PAH3;
DT 18-MAY-2010, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1998, sequence version 1.
DT 03-AUG-2022, entry version 102.
DE RecName: Full=Formate dehydrogenase {ECO:0000255|HAMAP-Rule:MF_03210, ECO:0000312|EMBL:CAB54834.1};
DE Short=FDH {ECO:0000255|HAMAP-Rule:MF_03210};
DE EC=1.17.1.9 {ECO:0000255|HAMAP-Rule:MF_03210, ECO:0000269|PubMed:10691964, ECO:0000269|PubMed:11171126, ECO:0000269|PubMed:1248477, ECO:0000269|PubMed:17525463, ECO:0000269|PubMed:9226256};
DE AltName: Full=NAD-dependent formate dehydrogenase {ECO:0000255|HAMAP-Rule:MF_03210, ECO:0000312|EMBL:AAC49766.1};
GN Name=FDH1 {ECO:0000303|PubMed:9226256, ECO:0000312|EMBL:AAC49766.1};
GN Synonyms=FDH {ECO:0000312|EMBL:CAA09466.2},
GN FDH3 {ECO:0000312|EMBL:CAB54834.1};
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], AND PROTEIN SEQUENCE OF 1-25.
RC STRAIN=ATCC 56294 / CBS 8030 / CCRC 21757 / NRRL Y-17325;
RX PubMed=7557425; DOI=10.1016/0378-1119(95)00347-9;
RA Allen S.J., Holbrook J.J.;
RT "Isolation, sequence and overexpression of the gene encoding NAD-dependent
RT formate dehydrogenase from the methylotrophic yeast Candida methylica.";
RL Gene 162:99-104(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 2-45; 57-76; 87-103;
RP 190-201; 207-236; 242-246; 292-326 AND 329-354, FUNCTION, CATALYTIC
RP ACTIVITY, DISRUPTION PHENOTYPE, AND INDUCTION.
RC STRAIN=S2 {ECO:0000312|EMBL:AAC49766.1};
RX PubMed=9226256; DOI=10.1128/jb.179.14.4480-4485.1997;
RA Sakai Y., Murdanoto A.P., Konishi T., Iwamatsu A., Kato N.;
RT "Regulation of the formate dehydrogenase gene, FDH1, in the methylotrophic
RT yeast Candida boidinii and growth characteristics of an FDH1-disrupted
RT strain on methanol, methylamine, and choline.";
RL J. Bacteriol. 179:4480-4485(1997).
RN [3]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 1-15, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, BIOTECHNOLOGY, AND MUTAGENESIS OF CYS-23 AND
RP CYS-262.
RC STRAIN=ATCC 32195 {ECO:0000312|EMBL:CAB54834.1};
RX PubMed=10691964; DOI=10.1046/j.1432-1327.2000.01123.x;
RA Slusarczyk H., Felber S., Kula M.R., Pohl M.;
RT "Stabilization of NAD-dependent formate dehydrogenase from Candida boidinii
RT by site-directed mutagenesis of cysteine residues.";
RL Eur. J. Biochem. 267:1280-1289(2000).
RN [4]
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 1-30 AND 132-140, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF PHE-69;
RP ASN-119; ILE-175; GLN-197; ARG-258; GLN-287; PRO-288 AND HIS-311.
RC STRAIN=NCYC 1513 {ECO:0000312|EMBL:CAA09466.2};
RX PubMed=11171126; DOI=10.1042/0264-6021:3540455;
RA Labrou N.E., Rigden D.J.;
RT "Active-site characterization of Candida boidinii formate dehydrogenase.";
RL Biochem. J. 354:455-463(2001).
RN [5]
RP NUCLEOTIDE SEQUENCE [MRNA].
RC STRAIN=2.2159 {ECO:0000312|EMBL:ABE69165.2};
RA Zhang G., Yang G., Cao Z., Liu M.;
RL Submitted (JUL-2007) to the EMBL/GenBank/DDBJ databases.
RN [6]
RP PROTEIN SEQUENCE OF 357-363, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS
RP OF LYS-360.
RX PubMed=11054119; DOI=10.1046/j.1432-1327.2000.01761.x;
RA Labrou N.E., Rigden D.J., Clonis Y.D.;
RT "Characterization of the NAD+ binding site of Candida boidinii formate
RT dehydrogenase by affinity labelling and site-directed mutagenesis.";
RL Eur. J. Biochem. 267:6657-6664(2000).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RX PubMed=1248477; DOI=10.1111/j.1432-1033.1976.tb10108.x;
RA Schute H., Flossdorf J., Sahm H., Kula M.R.;
RT "Purification and properties of formaldehyde dehydrogenase and formate
RT dehydrogenase from Candida boidinii.";
RL Eur. J. Biochem. 62:151-160(1976).
RN [8]
RP BIOTECHNOLOGY.
RX DOI=10.1002/ceat.270170211;
RA Weuster-Botz D., Paschold H., Striegel B., Gieren H., Kula M.R.,
RA Wandrey C.;
RT "Continuous computer controlled production of formate dehydrogenase (FDH)
RT and isolation on a pilot scale.";
RL Chem. Eng. Technol. 17:131-137(1994).
RN [9]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF MUTANT GLU-47 AND (1.55 ANGSTROMS)
RP OF MUTANT VAL-328, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBUNIT, CATALYTIC AND COENZYME-BINDING REGIONS, AND MUTAGENESIS OF LYS-47
RP AND LYS-328.
RX PubMed=17525463; DOI=10.1110/ps.062741707;
RA Schirwitz K., Schmidt A., Lamzin V.S.;
RT "High-resolution structures of formate dehydrogenase from Candida
RT boidinii.";
RL Protein Sci. 16:1146-1156(2007).
CC -!- FUNCTION: Catalyzes the NAD(+)-dependent oxidation of formate to carbon
CC dioxide. Formate oxidation is the final step in the methanol oxidation
CC pathway in methylotrophic microorganisms. Has a role in the
CC detoxification of exogenous formate in non-methylotrophic organisms.
CC {ECO:0000255|HAMAP-Rule:MF_03210, ECO:0000269|PubMed:1248477,
CC ECO:0000269|PubMed:9226256}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=formate + NAD(+) = CO2 + NADH; Xref=Rhea:RHEA:15985,
CC ChEBI:CHEBI:15740, ChEBI:CHEBI:16526, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945; EC=1.17.1.9; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_03210, ECO:0000269|PubMed:10691964,
CC ECO:0000269|PubMed:11171126, ECO:0000269|PubMed:1248477,
CC ECO:0000269|PubMed:17525463, ECO:0000269|PubMed:9226256};
CC -!- ACTIVITY REGULATION: Cu(2+), Hg and p-chloromercuribenzoate are strong
CC inhibitors of enzyme activity and Ca(2+), Mg(2+), Zn(2+), Mn(2+),
CC Cd(2+) and Sn(2+) have no effect on activity indicating a cysteine
CC residue in the protein is essential for enzyme activity or to maintain
CC the proper structure of the enzyme. Nitrite and nitrate inhibit some
CC enzyme activity, however cyanide, azide, thiocyanate and cyanate are
CC strong inhibitors of the enzymatic reaction. The inhibition of cyanide
CC is competitive with formate and reversible.
CC {ECO:0000269|PubMed:1248477}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=13 mM for formate (at 30 degrees Celsius and at pH 7.5)
CC {ECO:0000269|PubMed:1248477};
CC KM=0.09 mM for NAD (at 30 degrees Celsius and at pH 7.5)
CC {ECO:0000269|PubMed:1248477};
CC KM=5.6 mM for formate (at 30 degrees Celsius and at pH 7.5)
CC {ECO:0000269|PubMed:10691964};
CC KM=0.045 mM for NAD (at 30 degrees Celsius and at pH 7.5)
CC {ECO:0000269|PubMed:10691964};
CC KM=2.42 mM for formate (at 25 degrees Celsius and at pH 7.5)
CC {ECO:0000269|PubMed:11171126};
CC KM=0.04 mM for NAD (at 25 degrees Celsius and at pH 7.5)
CC {ECO:0000269|PubMed:11171126};
CC KM=2.4 mM for formate (at 25 degrees Celsius and at pH 7.6)
CC {ECO:0000269|PubMed:11054119};
CC KM=0.04 mM for NAD (at 25 degrees Celsius and at pH 7.6)
CC {ECO:0000269|PubMed:11054119};
CC KM=20.0 mM for formate (at 20 degrees Celsius, at pH 7.5 and after 2
CC weeks of storage at 4 degrees Celsius in GF buffer)
CC {ECO:0000269|PubMed:17525463};
CC KM=0.05 mM for NAD (at 20 degrees Celsius, at pH 7.5 and after 2
CC weeks of storage at 4 degrees Celsius in GF buffer)
CC {ECO:0000269|PubMed:17525463};
CC KM=35.0 mM for formate (at 20 degrees Celsius, at pH 7.5 and after 4
CC months of storage at 4 degrees Celsius in GF buffer)
CC {ECO:0000269|PubMed:17525463};
CC KM=0.09 mM for NAD (at 20 degrees Celsius, at pH 7.5 and after 4
CC months of storage at 4 degrees Celsius in GF buffer)
CC {ECO:0000269|PubMed:17525463};
CC Vmax=6 uM/min/mg enzyme {ECO:0000269|PubMed:10691964};
CC pH dependence:
CC Optimum pH is 7.5-8.5. {ECO:0000269|PubMed:10691964,
CC ECO:0000269|PubMed:11054119, ECO:0000269|PubMed:11171126,
CC ECO:0000269|PubMed:1248477, ECO:0000269|PubMed:17525463};
CC Temperature dependence:
CC Broad temperature optima between 45 and 55 degrees Celsius. Reaction
CC rate increases steeply up to 55 degrees Celsius. 50% of activity lost
CC after incubation for 20 minutes at 57 degrees Celsius. Thermal
CC stability increases in the presence of glycerol.
CC {ECO:0000269|PubMed:10691964, ECO:0000269|PubMed:11054119,
CC ECO:0000269|PubMed:11171126, ECO:0000269|PubMed:1248477,
CC ECO:0000269|PubMed:17525463};
CC -!- SUBUNIT: Homodimer. {ECO:0000255|HAMAP-Rule:MF_03210,
CC ECO:0000269|PubMed:1248477, ECO:0000269|PubMed:17525463}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_03210}.
CC -!- INDUCTION: Expression is strongly induced by methanol, but is
CC completely repressed in the presence of glucose. However, methanol
CC induced expression is equally strong in cells grown on glucose when
CC formate, methylamine or choline is added. No expression is detected in
CC cells grown on glycerol. When formate, methylamine or choline is added
CC to the culture medium of glycerol- or glucose-grown cells, they exhibit
CC an induction of FDH1 expression. {ECO:0000269|PubMed:9226256}.
CC -!- DISRUPTION PHENOTYPE: Is able to grow on methanol in a batch culture
CC experiment, but its growth is greatly inhibited and a toxic level of
CC formate accumulates in the medium. Formate is not detected in the
CC medium in a methanol-limited chemostat culture but deletion mutant
CC shows only one-fourth of the growth yield of the wild-type.
CC {ECO:0000269|PubMed:9226256}.
CC -!- BIOTECHNOLOGY: Ideal catalyst for synthesizing chiral compounds of high
CC enantiomeric purity from prochiral precursors due to a favorable
CC thermodynamic equilibrium, the oxidation of formate to carbon dioxide
CC while also reducing NAD to NADH. However, the necessesity for the
CC presence of large quantities of the enzyme and its rapid inactivation
CC under biotransformation conditions results in higher costs for the
CC biocatalyst industry. In order to make this enzymatic reduction viable
CC and to perform it on a larger scale a more efficient and cost effective
CC process has been established. Site-directed mutagenesis has been
CC effective in stabilizing this commercially important enzyme for its
CC application in the biotransformation of trimethyl pyruvate to L-tert
CC leucine. {ECO:0000269|PubMed:10691964, ECO:0000269|Ref.8}.
CC -!- SIMILARITY: Belongs to the D-isomer specific 2-hydroxyacid
CC dehydrogenase family. FDH subfamily. {ECO:0000255|HAMAP-Rule:MF_03210}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; X81129; CAA57036.1; -; Genomic_DNA.
DR EMBL; AF004096; AAC49766.1; -; Genomic_DNA.
DR EMBL; AJ245934; CAB54834.1; -; Genomic_DNA.
DR EMBL; AJ011046; CAA09466.2; -; Genomic_DNA.
DR EMBL; DQ458777; ABE69165.2; -; Genomic_DNA.
DR PIR; JC4252; JC4252.
DR PDB; 2FSS; X-ray; 1.70 A; A/B/C/D=2-364.
DR PDB; 2J6I; X-ray; 1.55 A; A/B/C/D=2-364.
DR PDBsum; 2FSS; -.
DR PDBsum; 2J6I; -.
DR AlphaFoldDB; O13437; -.
DR SMR; O13437; -.
DR OrthoDB; 700058at2759; -.
DR BioCyc; MetaCyc:MON-17206; -.
DR BRENDA; 1.17.1.9; 1100.
DR SABIO-RK; O13437; -.
DR EvolutionaryTrace; O13437; -.
DR GO; GO:0005829; C:cytosol; ISS:UniProtKB.
DR GO; GO:0009326; C:formate dehydrogenase complex; IEA:UniProtKB-EC.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0008863; F:formate dehydrogenase (NAD+) activity; IDA:UniProtKB.
DR GO; GO:0070403; F:NAD+ binding; IDA:UniProtKB.
DR GO; GO:0016616; F:oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor; IEA:InterPro.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:0042426; P:choline catabolic process; IMP:UniProtKB.
DR GO; GO:0042183; P:formate catabolic process; IDA:UniProtKB.
DR GO; GO:0015946; P:methanol oxidation; IMP:UniProtKB.
DR GO; GO:0030416; P:methylamine metabolic process; IMP:UniProtKB.
DR GO; GO:0006734; P:NADH metabolic process; IMP:UniProtKB.
DR GO; GO:0006735; P:NADH regeneration; IDA:UniProtKB.
DR CDD; cd05302; FDH; 1.
DR HAMAP; MF_03210; Formate_dehydrogenase; 1.
DR InterPro; IPR006139; D-isomer_2_OHA_DH_cat_dom.
DR InterPro; IPR029753; D-isomer_DH_CS.
DR InterPro; IPR029752; D-isomer_DH_CS1.
DR InterPro; IPR006140; D-isomer_DH_NAD-bd.
DR InterPro; IPR033689; FDH_NAD-dep.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF00389; 2-Hacid_dh; 1.
DR Pfam; PF02826; 2-Hacid_dh_C; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR PROSITE; PS00065; D_2_HYDROXYACID_DH_1; 1.
DR PROSITE; PS00670; D_2_HYDROXYACID_DH_2; 1.
DR PROSITE; PS00671; D_2_HYDROXYACID_DH_3; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Cytoplasm; Direct protein sequencing; NAD;
KW Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..364
FT /note="Formate dehydrogenase"
FT /id="PRO_0000393949"
FT BINDING 93
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 119
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 174..175
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 195
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 230..234
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 256
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 282
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT BINDING 311..314
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210"
FT SITE 258
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210,
FT ECO:0000305|PubMed:17525463"
FT SITE 311
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_03210,
FT ECO:0000305|PubMed:17525463"
FT VARIANT 9
FT /note="D -> G (in strain: 2.2159)"
FT /evidence="ECO:0000305, ECO:0000312|EMBL:ABE69165.2"
FT VARIANT 50..51
FT /note="ET -> GN (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 53
FT /note="E -> V (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 56
FT /note="K -> Q (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 79
FT /note="L -> I (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 84
FT /note="N -> K (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 87
FT /note="L -> S (in strain: ATCC 56294 / CBS 8030 / CCRC
FT 21757 / NRRL Y-17325)"
FT /evidence="ECO:0000269|PubMed:7557425"
FT VARIANT 108
FT /note="K -> R (in strain: 2.2159)"
FT /evidence="ECO:0000305, ECO:0000312|EMBL:ABE69165.2"
FT VARIANT 145
FT /note="I -> N (in strain: 2.2159)"
FT /evidence="ECO:0000305, ECO:0000312|EMBL:ABE69165.2"
FT VARIANT 184
FT /note="L -> V (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 202
FT /note="E -> D (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT VARIANT 308
FT /note="M -> T (in strain: 2.2159)"
FT /evidence="ECO:0000305, ECO:0000312|EMBL:ABE69165.2"
FT VARIANT 325
FT /note="E -> Q (in strain: 2.2159 and NCYC 1513)"
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 23
FT /note="C->S: Slight increase in substrate affinity for
FT formate but no change in affinity for NAD, 9 degrees
FT Celsius decrease in thermal stability compared to the wild-
FT type, significantly higher stability compared to wild-type
FT under biotransformation conditions, significantly more
FT stable in the presence of CuCl(2); when associated with A-
FT 262. Large increase in substrate affinity for formate but
FT no significant change in affinity for NAD, 13 degrees
FT Celsius decrease in thermal stability compared to the wild-
FT type, significantly more stable in the presence of CuCl(2);
FT when associated with V-262. No significant change in
FT affinity for formate or NAD, 5 degrees Celsius decrease in
FT thermal stability compared to the wild-type, significantly
FT higher stability compared to wild-type under
FT biotransformation conditions, and significantly more stable
FT in the presence of CuCl(2)."
FT /evidence="ECO:0000269|PubMed:10691964"
FT MUTAGEN 47
FT /note="K->E: Slight increase in substrate affinity for
FT formate and also affinity for NAD increases by half after 2
FT weeks. Also after 4 months affinity for formate increases
FT by more than half and affinity for NAD increases by more
FT than half. Retains 84% of residual activity after
FT incubation for 20 minutes at a thermal inactivation
FT temperature of 55 degrees Celsius in samples stored for 2
FT weeks compared to wild-type which loses 50% of its activity
FT at 55 degrees Celsius."
FT /evidence="ECO:0000269|PubMed:17525463"
FT MUTAGEN 69
FT /note="F->A: 2-fold decrease in substrate affinity for
FT formate, but no significant change in affinity for NAD. A
FT significant reduction in catalytic activity compared to the
FT wild-type."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 119
FT /note="N->A: 94-fold decrease in substrate affinity for
FT formate and 2700-fold decrease in substrate affinity for
FT NAD. A significant reduction in catalytic activity compared
FT to the wild-type; when associated with A-311."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 119
FT /note="N->H: 80-fold decrease in substrate affinity for
FT formate and a 1250-fold decrease in substrate affinity for
FT NAD. A significant reduction in catalytic activity compared
FT to the wild-type."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 175
FT /note="I->A: 2-fold decrease in substrate affinity for
FT formate and a 12-fold decrease in substrate affinity for
FT NAD. A significant reduction in catalytic activity compared
FT to the wild-type."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 197
FT /note="Q->L: 4-fold decrease in substrate affinity for
FT formate but no significant change in affinity for NAD
FT compared to the wild-type."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 258
FT /note="R->A: No catalytic activity."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 262
FT /note="C->A: Slight increase in substrate affinity for
FT formate but no change in affinity for NAD, 9 degrees
FT Celsius decrease in thermal stability compared to the wild-
FT type, greater stability at a higher pH compared to the
FT wild-type; when associated with S-23."
FT /evidence="ECO:0000269|PubMed:10691964"
FT MUTAGEN 262
FT /note="C->V: Large increase in substrate affinity for
FT formate but no significant change in affinity for NAD, 13
FT degrees Celsius decrease in thermal stability compared to
FT the wild-type; when associated with S-23. Great increase in
FT substrate affinity for formate and NAD and 8 degrees
FT Celsius decrease in thermal stability compared to the wild-
FT type."
FT /evidence="ECO:0000269|PubMed:10691964"
FT MUTAGEN 287
FT /note="Q->A: 2-fold decrease in substrate affinity for
FT formate and 3-fold decrease in substrate affinity for NAD
FT compared to the wild-type; when associated with A-311."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 287
FT /note="Q->E: 380-fold decrease in substrate affinity for
FT formate and 3-fold decrease in substrate affinity for NAD
FT compared to the wild-type; when associated with T-288. No
FT significant decrease in substrate affinity for formate but
FT a 4-fold decrease in substrate affinity for NAD and a
FT significant reduction in catalytic activity compared to the
FT wild-type, a more acidic pH is seen than in the wild-type,
FT preventing formate binding by a single ionization of a
FT group compared to that of the wild-type."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 288
FT /note="P->T: 380-fold decrease in substrate affinity for
FT formate and 3-fold decrease in substrate affinity for NAD
FT compared to the wild-type; when associated with E-287."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 311
FT /note="H->A: 2-fold decrease in substrate affinity for
FT formate and 3-fold decrease in substrate affinity for NAD
FT compared to the wild-type; when associated with A-287. 93-
FT fold decrease in substrate affinity for formate and 2700-
FT fold decrease in substrate affinity for NAD, and a
FT significant reduction in catalytic activity compared to the
FT wild-type; when associated with A-119."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 311
FT /note="H->Q: 10-fold decrease in substrate affinity for
FT formate and significant reduction in the catalytic activity
FT compared to the wild-type."
FT /evidence="ECO:0000269|PubMed:11171126"
FT MUTAGEN 328
FT /note="K->V: A 75% increase in substrate affinity for
FT formate after 2 weeks and a 50% increase in affinity for
FT NAD. However, after 4 months the affinity for formate
FT increases 7-fold and affinity for NAD increases by 2
FT thirds. Retains 70% of residual activity after incubation
FT for 20 minutes at a thermal inactivation temperature of 55
FT degrees Celsius in samples stored for 2 weeks compared to
FT wild-type which loses 50% of its activity at 55 degrees
FT Celsius."
FT /evidence="ECO:0000269|PubMed:17525463"
FT MUTAGEN 360
FT /note="K->A: Exhibits no change in substrate affinity for
FT formate, but shows a 4-fold decrease in substrate affinity
FT for NAD implying that L-360 side chain forms strong
FT interactions with the cofactor. A higher reaction rate is
FT observed at an acidic and basic pH values."
FT /evidence="ECO:0000269|PubMed:11054119"
FT CONFLICT 19..23
FT /note="KLYGC -> EKLYG (in Ref. 4; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 23
FT /note="C -> T (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 2..6
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 12..16
FT /evidence="ECO:0007829|PDB:2J6I"
FT TURN 24..26
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 27..29
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 31..36
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 40..45
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 49..52
FT /evidence="ECO:0007829|PDB:2FSS"
FT HELIX 53..57
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 58..60
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 62..66
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 76..81
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 87..93
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 100..106
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 111..114
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 120..135
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 138..146
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 152..156
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 166..170
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 174..183
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 184..186
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 189..194
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 201..206
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 209..211
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 215..220
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 223..227
FT /evidence="ECO:0007829|PDB:2J6I"
FT TURN 233..237
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 241..244
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 251..255
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 259..261
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 264..272
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 275..282
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 285..288
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 294..297
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 306..308
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 313..315
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 317..335
FT /evidence="ECO:0007829|PDB:2J6I"
FT HELIX 343..345
FT /evidence="ECO:0007829|PDB:2J6I"
FT STRAND 346..349
FT /evidence="ECO:0007829|PDB:2J6I"
SQ SEQUENCE 364 AA; 40370 MW; 1B30982E0D5B77E8 CRC64;
MKIVLVLYDA GKHAADEEKL YGCTENKLGI ANWLKDQGHE LITTSDKEGE TSELDKHIPD
ADIIITTPFH PAYITKERLD KAKNLKLVVV AGVGSDHIDL DYINQTGKKI SVLEVTGSNV
VSVAEHVVMT MLVLVRNFVP AHEQIINHDW EVAAIAKDAY DIEGKTIATI GAGRIGYRVL
ERLLPFNPKE LLYYDYQALP KEAEEKVGAR RVENIEELVA QADIVTVNAP LHAGTKGLIN
KELLSKFKKG AWLVNTARGA ICVAEDVAAA LESGQLRGYG GDVWFPQPAP KDHPWRDMRN
KYGAGNAMTP HYSGTTLDAQ TRYAEGTKNI LESFFTGKFD YRPQDIILLN GEYVTKAYGK
HDKK
