MNO_AMYME
ID MNO_AMYME Reviewed; 429 AA.
AC Q9RCG0;
DT 07-JAN-2015, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-2000, sequence version 1.
DT 25-MAY-2022, entry version 44.
DE RecName: Full=Methanol:N,N-dimethyl-4-nitrosoaniline oxidoreductase {ECO:0000303|PubMed:8449887};
DE Short=MNO {ECO:0000303|PubMed:8449887};
DE EC=1.1.99.37 {ECO:0000269|PubMed:8449887, ECO:0000269|Ref.3};
DE AltName: Full=Methanol dehydrogenase (nicotinoprotein) {ECO:0000305};
DE AltName: Full=Methanol:NDMA oxidoreductase {ECO:0000303|PubMed:8449887};
GN Name=mno;
OS Amycolatopsis methanolica.
OC Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae;
OC Amycolatopsis; Amycolatopsis methanolica group.
OX NCBI_TaxID=1814;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA Hektor H.J., Bron D., Dijkhuizen L.;
RL Submitted (FEB-1997) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP PROTEIN SEQUENCE OF 2-47, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND
RP SUBUNIT.
RX PubMed=8449887; DOI=10.1128/jb.175.6.1814-1822.1993;
RA Bystrykh L.V., Vonck J., van Bruggen E.F., van Beeumen J., Samyn B.,
RA Govorukhina N.I., Arfman N., Duine J.A., Dijkhuizen L.;
RT "Electron microscopic analysis and structural characterization of novel
RT NADP(H)-containing methanol: N,N'-dimethyl-4-nitrosoaniline oxidoreductases
RT from the Gram-positive methylotrophic bacteria Amycolatopsis methanolica
RT and Mycobacterium gastri MB19.";
RL J. Bacteriol. 175:1814-1822(1993).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, AND SUBSTRATE SPECIFICITY.
RC STRAIN=DSM 44096 / JCM 8087 / NBRC 15065 / NCIMB 11946 / NRRL B-24139 / LMD
RC 80.32 / 239;
RA Bystrykh L.V., Govorukhina N.I., van Ophem P.W., Hektor H.J.,
RA Dijkhuizen L., Duine J.A.;
RT "Formaldehyde dismutase activities in Gram-positive bacteria oxidizing
RT methanol.";
RL (er) J Gen Microbiol 139, 1979-1985(1993).
RN [4]
RP FUNCTION, DISRUPTION PHENOTYPE, AND INDUCTION.
RC STRAIN=DSM 44096 / JCM 8087 / NBRC 15065 / NCIMB 11946 / NRRL B-24139 / LMD
RC 80.32 / 239;
RX DOI=10.1016/0378-1097(96)00341-2;
RA Hektor H.J., Dijkhuizen L.;
RT "Mutational analysis of primary alcohol metabolism in the methylotrophic
RT actinomycete Amycolatopsis methanolica.";
RL (er) FEMS Microbiol Lett 144, 73-79(1996).
CC -!- FUNCTION: Catalyzes the oxidation of methanol to yield formaldehyde.
CC While the in vivo electron acceptor is not known, N,N-dimethyl-4-
CC nitrosoaniline (NDMA) can serve this function in vitro and is reduced
CC to 4-(hydroxylamino)-N,N-dimethylaniline. It can also use various other
CC primary alcohols, polyols and formaldehyde. In addition, MNO is able to
CC produce methylformate from methanol plus formaldehyde, and possesses a
CC formaldehyde dismutase and a NADH-dependent formaldehyde reductase
CC activity. {ECO:0000269|PubMed:8449887, ECO:0000269|Ref.1,
CC ECO:0000269|Ref.3, ECO:0000269|Ref.4}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=A + methanol = AH2 + formaldehyde; Xref=Rhea:RHEA:33571,
CC ChEBI:CHEBI:13193, ChEBI:CHEBI:16842, ChEBI:CHEBI:17499,
CC ChEBI:CHEBI:17790; EC=1.1.99.37;
CC Evidence={ECO:0000269|PubMed:8449887, ECO:0000269|Ref.3};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:8449887};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:8449887};
CC -!- COFACTOR:
CC Name=NADPH; Xref=ChEBI:CHEBI:57783;
CC Evidence={ECO:0000269|PubMed:8449887};
CC -!- ACTIVITY REGULATION: Inhibited by azide and hydrazine.
CC {ECO:0000269|Ref.3}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1 uM for isobutanol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=2 uM for butanol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=2 uM for propanol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=7 uM for formaldehyde (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=10 uM for 2-methoxyethanol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=18 uM for NDMA (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=47 uM for ethyleneglycol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=57 uM for ethanol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=110 uM for propan-2-ol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=970 uM for glycerol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=2.65 mM for methanol (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC KM=17 mM for ethanolamine (at pH 6.3 and 30 degrees Celsius)
CC {ECO:0000269|Ref.3};
CC Vmax=40 umol/min/mg enzyme with ethanolamine as substrate (at pH 6.3
CC and 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=51 umol/min/mg enzyme with formaldehyde as substrate (at pH 6.3
CC and 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=57 umol/min/mg enzyme with methanol as substrate (at pH 6.3 and
CC 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=63 umol/min/mg enzyme with 2-methoxyethanol as substrate (at pH
CC 6.3 and 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=65 umol/min/mg enzyme with NDMA as substrate (at pH 6.3 and 30
CC degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=66 umol/min/mg enzyme with ethyleneglycol as substrate (at pH
CC 6.3 and 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=72 umol/min/mg enzyme with glycerol as substrate (at pH 6.3 and
CC 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=73 umol/min/mg enzyme with isobutanol as substrate (at pH 6.3
CC and 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=73 umol/min/mg enzyme with propan-2-ol as substrate (at pH 6.3
CC and 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=76 umol/min/mg enzyme with butanol as substrate (at pH 6.3 and
CC 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=77 umol/min/mg enzyme with propanol as substrate (at pH 6.3 and
CC 30 degrees Celsius) {ECO:0000269|Ref.3};
CC Vmax=90 umol/min/mg enzyme with ethanol as substrate (at pH 6.3 and
CC 30 degrees Celsius) {ECO:0000269|Ref.3};
CC pH dependence:
CC Optimum pH is between 4.5 and 5. {ECO:0000269|Ref.3};
CC Temperature dependence:
CC The MNO activity increases with temperature up to 55 degrees Celsius.
CC {ECO:0000269|Ref.3};
CC -!- SUBUNIT: Homodecamer. {ECO:0000269|PubMed:8449887}.
CC -!- INDUCTION: By methanol and ethanol. {ECO:0000269|Ref.4}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene are unable to grow
CC methanol, ethanol, propan-1-ol and butan-1-ol, but are still able to
CC grow on formaldehyde. {ECO:0000269|Ref.4}.
CC -!- SIMILARITY: Belongs to the iron-containing alcohol dehydrogenase
CC family. {ECO:0000305}.
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DR EMBL; U89273; AAF21473.1; -; Genomic_DNA.
DR AlphaFoldDB; Q9RCG0; -.
DR SMR; Q9RCG0; -.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0070402; F:NADPH binding; IDA:UniProtKB.
DR GO; GO:0016491; F:oxidoreductase activity; IDA:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0015946; P:methanol oxidation; IDA:UniProtKB.
DR InterPro; IPR001670; ADH_Fe/GldA.
DR InterPro; IPR039697; Alcohol_dehydrogenase_Fe.
DR InterPro; IPR026338; NDMA_methanol_DH.
DR PANTHER; PTHR11496; PTHR11496; 1.
DR Pfam; PF00465; Fe-ADH; 1.
DR TIGRFAMs; TIGR04266; NDMA_methanol; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Magnesium; Methanol utilization; NADP;
KW Oxidoreductase; Zinc.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:8449887"
FT CHAIN 2..429
FT /note="Methanol:N,N-dimethyl-4-nitrosoaniline
FT oxidoreductase"
FT /id="PRO_0000431560"
SQ SEQUENCE 429 AA; 46843 MW; 8E462A5EFD4D32D2 CRC64;
MQVDELLKPF PIKEFHPFPR ALLGPGAHEM IGPEALKLGF KKTLVMTSGL RGSDIVHKIT
ESMKYHGLEV VLYDKVESNP KDYNVMDAVK LYQENKCDSF VSIGGGSSHD ACKGARISVA
HDGRNVNDFE GFNKSENPRN PPHIAVSTTA GTGSETSWAY VITDTTTDPD NPHKYVAFDD
ASVATLAIDD PVLYYSCPID YTAQCGFDVL AHASEPYVSR LNFEPSLGNA LRAIKLTAEN
LRQATWNPSE LSGREGMMYA QYIAAQAFNS GGLGIIHSIS HAVSAFYDTH HGLNNAIALP
RVWAFNMPVA YKRFADMAEA MGVDTHGMTD VQAADALAAA IRLLRDVGIP EKFTDVTQDS
YSKNRLGQGP TKFYEQASVI KGDDEDVDRI TNHVLGDACT PGNAKECTFE TVRPVVDHCM
NGDLDDLLS
