MEDH_BACMT
ID MEDH_BACMT Reviewed; 381 AA.
AC P31005;
DT 01-JUL-1993, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 75.
DE RecName: Full=NAD-dependent methanol dehydrogenase {ECO:0000303|PubMed:1995642};
DE Short=MDH {ECO:0000303|PubMed:1995642};
DE Short=MEDH {ECO:0000303|PubMed:2673121};
DE EC=1.1.1.244 {ECO:0000269|PubMed:12089158, ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1995643, ECO:0000269|PubMed:2673121};
DE AltName: Full=Type 3 alcohol dehydrogenase {ECO:0000303|PubMed:1644761};
GN Name=mdh {ECO:0000303|PubMed:2673121};
OS Bacillus methanolicus.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=1471;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-15, FUNCTION,
RP COFACTOR, AND ACTIVITY REGULATION.
RC STRAIN=C1;
RX PubMed=1644761; DOI=10.1128/jb.174.16.5346-5353.1992;
RA de Vries G.E., Arfman N., Terpstra P., Dijkhuizen L.;
RT "Cloning, expression, and sequence analysis of the Bacillus methanolicus C1
RT methanol dehydrogenase gene.";
RL J. Bacteriol. 174:5346-5353(1992).
RN [2]
RP PROTEIN SEQUENCE OF 2-61, FUNCTION, COFACTOR, ACTIVITY REGULATION, AND
RP SUBUNIT.
RC STRAIN=C1;
RX PubMed=1995642; DOI=10.1016/s0021-9258(19)67885-3;
RA Vonck J., Arfman N., de Vries G.E., van Beeumen J., van Bruggen E.F.J.,
RA Dijkhuizen L.;
RT "Electron microscopic analysis and biochemical characterization of a novel
RT methanol dehydrogenase from the thermotolerant Bacillus sp. C1.";
RL J. Biol. Chem. 266:3949-3954(1991).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, SUBUNIT, AND SUBSTRATE
RP SPECIFICITY.
RC STRAIN=C1;
RX PubMed=2673121; DOI=10.1007/bf00409664;
RA Arfman N., Watling E.M., Clement W., van Oosterwijk R.J., de Vries G.E.,
RA Harder W., Attwood M.M., Dijkhuizen L.;
RT "Methanol metabolism in thermotolerant methylotrophic Bacillus strains
RT involving a novel catabolic NAD-dependent methanol dehydrogenase as a key
RT enzyme.";
RL Arch. Microbiol. 152:280-288(1989).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY
RP REGULATION.
RC STRAIN=C1;
RX PubMed=1995643; DOI=10.1016/s0021-9258(19)67886-5;
RA Arfman N., Van Beeumen J., De Vries G.E., Harder W., Dijkhuizen L.;
RT "Purification and characterization of an activator protein for methanol
RT dehydrogenase from thermotolerant Bacillus spp.";
RL J. Biol. Chem. 266:3955-3960(1991).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND REACTION
RP MECHANISM.
RC STRAIN=C1;
RX PubMed=12089158; DOI=10.1074/jbc.m205617200;
RA Kloosterman H., Vrijbloed J.W., Dijkhuizen L.;
RT "Molecular, biochemical, and functional characterization of a Nudix
RT hydrolase protein that stimulates the activity of a nicotinoprotein alcohol
RT dehydrogenase.";
RL J. Biol. Chem. 277:34785-34792(2002).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF
RP GLY-13; GLY-15; ASP-88; GLY-95; SER-97; ASP-100 AND LYS-103, AND COFACTOR.
RC STRAIN=C1;
RX PubMed=12351635; DOI=10.1074/jbc.m207547200;
RA Hektor H.J., Kloosterman H., Dijkhuizen L.;
RT "Identification of a magnesium-dependent NAD(P)(H)-binding domain in the
RT nicotinoprotein methanol dehydrogenase from Bacillus methanolicus.";
RL J. Biol. Chem. 277:46966-46973(2002).
CC -!- FUNCTION: Catalyzes the oxidation of methanol to yield formaldehyde. It
CC possesses a NADH-dependent formaldehyde reductase activity and cannot
CC use NADP. {ECO:0000269|PubMed:12089158, ECO:0000269|PubMed:12351635,
CC ECO:0000269|PubMed:1644761, ECO:0000269|PubMed:1995642,
CC ECO:0000269|PubMed:1995643, ECO:0000269|PubMed:2673121}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=methanol + NAD(+) = formaldehyde + H(+) + NADH;
CC Xref=Rhea:RHEA:19401, ChEBI:CHEBI:15378, ChEBI:CHEBI:16842,
CC ChEBI:CHEBI:17790, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945;
CC EC=1.1.1.244; Evidence={ECO:0000269|PubMed:12089158,
CC ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1995643,
CC ECO:0000269|PubMed:2673121};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1644761,
CC ECO:0000269|PubMed:1995642, ECO:0000269|PubMed:2673121};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1644761,
CC ECO:0000269|PubMed:1995642};
CC -!- ACTIVITY REGULATION: Stimulated by the activator protein Act wich
CC requires the presence of magnesium ions. Inhibited by 1,10-
CC phenanthroline. {ECO:0000269|PubMed:1644761,
CC ECO:0000269|PubMed:1995642, ECO:0000269|PubMed:1995643}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=9 uM for NADH {ECO:0000269|PubMed:12351635};
CC KM=0.02 mM for NAD {ECO:0000269|PubMed:12089158,
CC ECO:0000269|PubMed:12351635};
CC KM=0.7 mM for formaldehyde (without activator protein at pH 9.5 and
CC 50 degrees Celsius) {ECO:0000269|PubMed:1995643};
CC KM=2 mM for formaldehyde (with activator protein at pH 9.5 and 50
CC degrees Celsius) {ECO:0000269|PubMed:1995643};
CC KM=57 mM for ethanol (with the activator protein at pH 9.5 and 50
CC degrees Celsius) {ECO:0000269|PubMed:1995643};
CC KM=94 mM for ethanol (without activator protein at pH 9.5 and 50
CC degrees Celsius) {ECO:0000269|PubMed:1995643};
CC KM=140 mM for methanol (with the activator protein at pH 9.5 and 50
CC degrees Celsius) {ECO:0000269|PubMed:1995643};
CC KM=230 mM for methanol (without activator protein at pH 9.5 and 50
CC degrees Celsius) {ECO:0000269|PubMed:1995643};
CC Vmax=1310 umol/min/mg enzyme {ECO:0000269|PubMed:12089158,
CC ECO:0000269|PubMed:12351635};
CC -!- PATHWAY: One-carbon metabolism; methanol degradation; formaldehyde from
CC methanol: step 1/1. {ECO:0000305}.
CC -!- SUBUNIT: Homodecamer. {ECO:0000269|PubMed:1995642,
CC ECO:0000269|PubMed:2673121}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:2673121}.
CC -!- SIMILARITY: Belongs to the iron-containing alcohol dehydrogenase
CC family. {ECO:0000305}.
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DR EMBL; M65004; AAA22593.1; -; Genomic_DNA.
DR AlphaFoldDB; P31005; -.
DR SMR; P31005; -.
DR KEGG; ag:AAA22593; -.
DR BioCyc; MetaCyc:MON-15636; -.
DR SABIO-RK; P31005; -.
DR UniPathway; UPA00928; UER00893.
DR GO; GO:0005737; C:cytoplasm; IDA:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0050093; F:methanol dehydrogenase activity; IDA:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0046170; P:methanol catabolic process; IEA:UniProtKB-UniPathway.
DR InterPro; IPR001670; ADH_Fe/GldA.
DR InterPro; IPR018211; ADH_Fe_CS.
DR InterPro; IPR039697; Alcohol_dehydrogenase_Fe.
DR PANTHER; PTHR11496; PTHR11496; 1.
DR Pfam; PF00465; Fe-ADH; 1.
DR PROSITE; PS00913; ADH_IRON_1; 1.
DR PROSITE; PS00060; ADH_IRON_2; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Direct protein sequencing; Magnesium; Methanol utilization; NAD;
KW Oxidoreductase; Zinc.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:1644761,
FT ECO:0000269|PubMed:1995642"
FT CHAIN 2..381
FT /note="NAD-dependent methanol dehydrogenase"
FT /id="PRO_0000087834"
FT MUTAGEN 13
FT /note="G->A: Shows a reduced dehydrogenase activity."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 15
FT /note="G->A: Shows almost the same dehydrogenase activity
FT as the wild-type."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 88
FT /note="D->N: Shows almost the same dehydrogenase activity
FT as the wild-type."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 95
FT /note="G->A: Shows a 10-fold decreased affinity for NAD and
FT NADH and a strongly reduced dehydrogenase activity.
FT Completely insensitive to the stimulating effect of the
FT activator protein Act."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 97
FT /note="S->G: Shows an increase of the dehydrogenase
FT activity and a decrease of the affinity for NAD and NADH.
FT Completely insensitive to the stimulating effect of the
FT activator protein Act. It does not bind NAD."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 97
FT /note="S->T: Shows an increase of the dehydrogenase
FT activity and affinity for NAD and NADH."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 100
FT /note="D->N: Loss of dehydrogenase activity. It still binds
FT NADH."
FT /evidence="ECO:0000269|PubMed:12351635"
FT MUTAGEN 103
FT /note="K->R: Loss of dehydrogenase activity. It does not
FT bind NADH."
FT /evidence="ECO:0000269|PubMed:12351635"
FT CONFLICT 18
FT /note="K -> L (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 42
FT /note="S -> A (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 50
FT /note="A -> D (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 59
FT /note="D -> N (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 381 AA; 40045 MW; 8EA251B36F73749D CRC64;
MTNFFIPPAS VIGRGAVKEV GTRLKQIGAK KALIVTDAFL HSTGLSEEVA KNIREAGLDV
AIFPKAQPDP ADTQVHEGVD VFKQENCDAL VSIGGGSSHD TAKAIGLVAA NGGRINDYQG
VNSVEKPVVP VVAITTTAGT GSETTSLAVI TDSARKVKMP VIDEKITPTV AIVDPELMVK
KPAGLTIATG MDALSHAIEA YVAKGATPVT DAFAIQAMKL INEYLPKAVA NGEDIEAREA
MAYAQYMAGV AFNNGGLGLV HSISHQVGGV YKLQHGICNS VNMPHVCAFN LIAKTERFAH
IAELLGENVS GLSTAAAAER AIVALERYNK NFGIPSGYAE MGVKEEDIEL LAKNAFEDVC
TQSNPRVATV QDIAQIIKNA L
