DNTB_BURSP
ID DNTB_BURSP Reviewed; 548 AA.
AC Q2PWU9; P71029;
DT 28-MAR-2018, integrated into UniProtKB/Swiss-Prot.
DT 24-JAN-2006, sequence version 1.
DT 03-AUG-2022, entry version 53.
DE RecName: Full=4-methyl-5-nitrocatechol 5-monooxygenase {ECO:0000303|PubMed:16751499};
DE Short=4M5NC monooxygenase {ECO:0000303|PubMed:16751499};
DE Short=MNC monooxygenase {ECO:0000303|PubMed:8830701};
DE EC=1.14.13.210 {ECO:0000269|PubMed:16751499, ECO:0000269|PubMed:8830701};
DE AltName: Full=4-methyl-5-nitrocatechol oxygenase {ECO:0000303|PubMed:8830701};
GN Name=dntB {ECO:0000303|PubMed:8830701};
OS Burkholderia sp.
OC Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales;
OC Burkholderiaceae; Burkholderia.
OX NCBI_TaxID=36773;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, COFACTOR, AND SUBUNIT.
RC STRAIN=DNT {ECO:0000312|EMBL:AAC44479.1};
RX PubMed=8830701; DOI=10.1128/jb.178.20.6019-6024.1996;
RA Haigler B.E., Suen W.C., Spain J.C.;
RT "Purification and sequence analysis of 4-methyl-5-nitrocatechol oxygenase
RT from Burkholderia sp. strain DNT.";
RL J. Bacteriol. 178:6019-6024(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF MET-22 AND LEU-380.
RC STRAIN=DNT {ECO:0000312|EMBL:ABC00744.1};
RX PubMed=16751499; DOI=10.1128/aem.02966-05;
RA Leungsakul T., Johnson G.R., Wood T.K.;
RT "Protein engineering of the 4-methyl-5-nitrocatechol monooxygenase from
RT Burkholderia sp. strain DNT for enhanced degradation of nitroaromatics.";
RL Appl. Environ. Microbiol. 72:3933-3939(2006).
CC -!- FUNCTION: Involved in the degradation of 2,4-dinitrotoluene (2,4-DNT).
CC Catalyzes the removal of the nitro group from 4-methyl-5-nitrocatechol
CC (MNC) to yield 2-hydroxy-5-methylquinone (PubMed:8830701,
CC PubMed:16751499). It can use both NADH and NADPH as electron donors,
CC but prefers NADPH (PubMed:8830701). Also able to use 4-nitrocatechol as
CC substrate (PubMed:8830701). {ECO:0000269|PubMed:16751499,
CC ECO:0000269|PubMed:8830701}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-methyl-5-nitrocatechol + NADPH + O2 = 2-hydroxy-5-
CC methylquinone + H(+) + H2O + NADP(+) + nitrite; Xref=Rhea:RHEA:48012,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16301, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:81666, ChEBI:CHEBI:88190; EC=1.14.13.210;
CC Evidence={ECO:0000269|PubMed:16751499, ECO:0000269|PubMed:8830701};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=4-methyl-5-nitrocatechol + NADH + O2 = 2-hydroxy-5-
CC methylquinone + H(+) + H2O + NAD(+) + nitrite; Xref=Rhea:RHEA:48016,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16301, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945,
CC ChEBI:CHEBI:81666, ChEBI:CHEBI:88190; EC=1.14.13.210;
CC Evidence={ECO:0000269|PubMed:16751499, ECO:0000269|PubMed:8830701};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:8830701};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:8830701};
CC -!- ACTIVITY REGULATION: Activated by magnesium or manganese ions.
CC Inhibited by concentrations of 4-methyl-5-nitrocatechol (MNC) above 2
CC mM. {ECO:0000269|PubMed:8830701}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=15.5 uM for NADPH {ECO:0000269|PubMed:8830701};
CC KM=50 uM for oxygen {ECO:0000269|PubMed:8830701};
CC KM=350 uM for 4-nitrophenol (4NP) {ECO:0000269|PubMed:16751499};
CC KM=1220 uM for NADH {ECO:0000269|PubMed:8830701};
CC Vmax=14 nmol/min/mg enzyme with 4-nitrophenol (4NP) as substrate
CC {ECO:0000269|PubMed:16751499};
CC Note=kcat is 0.014 sec(-1) with 4-nitrophenol (4NP) as substrate.
CC {ECO:0000269|PubMed:16751499};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:8830701}.
CC -!- SIMILARITY: Belongs to the PheA/TfdB FAD monooxygenase family.
CC {ECO:0000305}.
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; U68411; AAC44479.1; -; Genomic_DNA.
DR EMBL; DQ298257; ABC00744.1; -; Genomic_DNA.
DR AlphaFoldDB; Q2PWU9; -.
DR SMR; Q2PWU9; -.
DR KEGG; ag:AAC44479; -.
DR KEGG; ag:ABC00744; -.
DR BioCyc; MetaCyc:MON-13337; -.
DR BRENDA; 1.14.13.210; 1033.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW.
DR GO; GO:0019439; P:aromatic compound catabolic process; IEA:UniProtKB-KW.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR002938; FAD-bd.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR Pfam; PF01494; FAD_binding_3; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW Aromatic hydrocarbons catabolism; FAD; Flavoprotein; Monooxygenase; NAD;
KW NADP; Oxidoreductase.
FT CHAIN 1..548
FT /note="4-methyl-5-nitrocatechol 5-monooxygenase"
FT /id="PRO_0000443520"
FT MUTAGEN 22
FT /note="M->L: Able to accept the two new substrates 4-
FT nitrophenol (4NP) and 3-methyl-4-nitrophenol (3M4NP); when
FT associated with I-380."
FT /evidence="ECO:0000269|PubMed:16751499"
FT MUTAGEN 380
FT /note="L->I: Able to accept the two new substrates 4-
FT nitrophenol (4NP) and 3-methyl-4-nitrophenol (3M4NP); when
FT associated with L-22."
FT /evidence="ECO:0000269|PubMed:16751499"
FT CONFLICT 312
FT /note="A -> R (in Ref. 1; AAC44479)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 548 AA; 59083 MW; 846E65119A8F6593 CRC64;
MTRPLETPPD IEVPVLIVGG SMVGLSTALF LSHYGIQAMA VERHERTAIH PRAGHFHLRT
LELLRSVGLE EVVARTSAEA FFPNGGINAV QSLAGGETAS FISNLNAGVE EFSPTRRLFI
AQQALEPILR SRAEELGADL RYSTEVVSVV DDGEGVTTVI RDKASGQERT VRSRYLVASD
GWRSQRRAQL GIETRGQGLL SRSATIYFRA DCRELLAGTH LGVIYVLNER LRGFFRFEKS
LQSGFLGVAT LGDPTRPGAL DVSAGFTTDT AVELVRAAIG VPDIDVEIQD VAHWEATAAL
ADRYRGGRIF LAGDAAHVVP PYGGFGGNTG VQDAHNLASK LALVLDGTAG EALLDTYEAE
RRPVGALTVD QAFSRYIRRL APEFLDEQTP ELVDDFSMEL GYRYHSPAVL TEDDDKAVDQ
AVVGHPREAL GRPGSRAPHV ALRVDDHDRS VLDLLGRDFV VLAGPAGQVW AEAAERASKE
LGLPLSAYVV GSDTPVADVE GRFADAYGLS DAGVALVRPD GFIAWRSRDL AEDPEAALTD
ALRAVLCR
