BPHA_ASPNG
ID BPHA_ASPNG Reviewed; 517 AA.
AC P17549;
DT 01-AUG-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1990, sequence version 1.
DT 03-AUG-2022, entry version 110.
DE RecName: Full=Benzoate 4-monooxygenase bphA {ECO:0000303|PubMed:11594739};
DE EC=1.14.14.92 {ECO:0000269|PubMed:11594739};
DE AltName: Full=Benzoate-para-hydroxylase A {ECO:0000303|PubMed:11594739};
DE Short=BpH {ECO:0000303|PubMed:11594739};
DE AltName: Full=Cytochrome P450 monooxygenase cyp53A1 {ECO:0000303|PubMed:11594739};
GN Name=bphA {ECO:0000303|PubMed:1868576};
GN Synonyms=cyp53A1 {ECO:0000303|PubMed:11594739};
OS Aspergillus niger.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=5061;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 1015 / NV DSM 2061;
RX PubMed=2250647; DOI=10.1007/bf00265053;
RA van Gorcom R.F.M., Boschloo J.G., Kuijvenhoven A., Lange J., van Vark A.J.,
RA Bos C.J., van Balken J.A.M., Pouwels P.H., van den Hondel C.A.M.J.J.;
RT "Isolation and molecular characterisation of the benzoate-para-hydroxylase
RT gene (bphA) of Aspergillus niger: a member of a new gene family of the
RT cytochrome P450 superfamily.";
RL Mol. Gen. Genet. 223:192-197(1990).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA].
RA Higgins U.M., Darby R.M., Maddison A., Lamb D.C., Draper J.;
RT "Yeast heterologous expression of a benzoate 4-hydroxylase (CYP53) from
RT Aspergillus niger.";
RL Submitted (SEP-2001) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP IDENTIFICATION.
RX PubMed=1868576; DOI=10.1007/bf00355052;
RA Boschloo J.G., Moonen E., van Gorcom R.F., Hermes H.F., Bos C.J.;
RT "Genetic analysis of Aspergillus niger mutants defective in benzoate-4-
RT hydroxylase function.";
RL Curr. Genet. 19:261-264(1991).
RN [4]
RP INDUCTION.
RX PubMed=10852481; DOI=10.1007/s004380051207;
RA van den Brink J.M., Punt P.J., van Gorcom R.F., van den Hondel C.A.;
RT "Regulation of expression of the Aspergillus niger benzoate para-
RT hydroxylase cytochrome P450 system.";
RL Mol. Gen. Genet. 263:601-609(2000).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP CHARACTERIZATION.
RC STRAIN=T18.5;
RX PubMed=11594739; DOI=10.1006/abbi.2001.2534;
RA Faber B.W., van Gorcom R.F.M., Duine J.A.;
RT "Purification and characterization of benzoate-para-hydroxylase, a
RT cytochrome P450 (CYP53A1), from Aspergillus niger.";
RL Arch. Biochem. Biophys. 394:245-254(2001).
RN [6]
RP FUNCTION, INDUCTION, AND DISRUPTION PHENOTYPE.
RX DOI=10.1021/acssuschemeng.9b04918;
RA Lubbers R.J.M., Dilokpimol A., Peng M., Visser J., Makela M.R.,
RA Hilden K.S., de Vries R.P.;
RT "Discovery of novel p-hydroxybenzoate-m-hydroxylase, protocatechuate 3,4
RT ring-cleavage dioxygenase, and hydroxyquinol 1,2 ring-cleavage dioxygenase
RT from the filamentous fungus Aspergillus niger.";
RL ACS Sustain. Chem. Eng. 7:19081-19089(2019).
CC -!- FUNCTION: Cytochrome P450 monooxygenase; part of the benzoic acid
CC degradation pathway also known as the protocatechuic acid pathway
CC (PubMed:11594739, Ref.6). Benzoic acid debradation begins with the
CC conversion of benzoic acid into 4-hydroxybenzoic acid through
CC hydroxylation by the benzoate-4-monooxygenase bphA, and its partner
CC NADPH-cytochrome P450 reductase cprA which act as a mediator in
CC electron donation from NADPH (PubMed:11594739). 4-Hydroxybenzoic acid
CC is then converted into 3,4-dihydroxybenzoic acid (also called
CC protocatechuic acid) by the p-hydroxybenzoate-m-hydroxylase phhA
CC (Ref.6). Protocatechuic acid is converted into 3-carboxy-cis,cis-
CC muconic acid by the intradiol ring-cleavage dioxygenase prcA, which is
CC further metabolized through the 3-oxoadipate pathway to finally enter
CC the tricarboxylic acid cycle (TCA) (Ref.6).
CC {ECO:0000269|PubMed:11594739, ECO:0000269|Ref.6}.
CC -!- FUNCTION: Responsible for cytochrome P450 dependent benzoate
CC hydroxylation in microsomes; requires cprA as the mediator in electron
CC donation from NADPH. {ECO:0000269|PubMed:11594739}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=benzoate + O2 + reduced [NADPH--hemoprotein reductase] = 4-
CC hydroxybenzoate + H(+) + H2O + oxidized [NADPH--hemoprotein
CC reductase]; Xref=Rhea:RHEA:18033, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16150, ChEBI:CHEBI:17879, ChEBI:CHEBI:57618,
CC ChEBI:CHEBI:58210; EC=1.14.14.92;
CC Evidence={ECO:0000269|PubMed:11594739};
CC -!- COFACTOR:
CC Name=heme; Xref=ChEBI:CHEBI:30413;
CC Evidence={ECO:0000250|UniProtKB:P04798};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.083 mM for benzoate {ECO:0000269|PubMed:11594739};
CC KM=0.097 mM for 2-fluorobenzoate {ECO:0000269|PubMed:11594739};
CC KM=0.31 mM for 2-chlorobenzoate {ECO:0000269|PubMed:11594739};
CC KM=0.28 mM for 2-hydroxybenzoate {ECO:0000269|PubMed:11594739};
CC KM=1.6 mM for 2-methylbenzoate {ECO:0000269|PubMed:11594739};
CC KM=0.086 mM for 3-fluorobenzoate {ECO:0000269|PubMed:11594739};
CC KM=0.127 mM for 3-chlorobenzoate {ECO:0000269|PubMed:11594739};
CC KM=0.673 mM for 3-hydroxybenzoate {ECO:0000269|PubMed:11594739};
CC KM=0.189 mM for 3-methylbenzoate {ECO:0000269|PubMed:11594739};
CC Note=kcat is 270 min(-1) for benzoate. {ECO:0000269|PubMed:11594739};
CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC protein {ECO:0000255}.
CC -!- INDUCTION: Expression is induced in the presence of caffeic acid, p-
CC coumaric acid, p-hydroxybenzoic acid, protocatechuic acid, and benzoic
CC acid. {ECO:0000269|PubMed:10852481, ECO:0000269|Ref.6}.
CC -!- DISRUPTION PHENOTYPE: Leads to reduced growth on benzoic acid (Ref.6).
CC Growth is also reduced on benzaldehyde, benzyl alcohol, and cinnamic
CC acid (Ref.6). {ECO:0000269|Ref.6}.
CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}.
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DR EMBL; X52521; CAA36753.1; -; Genomic_DNA.
DR EMBL; AJ347748; CAC69995.1; -; mRNA.
DR PIR; S12015; S12015.
DR AlphaFoldDB; P17549; -.
DR SMR; P17549; -.
DR STRING; 5061.CADANGAP00007746; -.
DR VEuPathDB; FungiDB:An09g03500; -.
DR VEuPathDB; FungiDB:ASPNIDRAFT2_1148703; -.
DR VEuPathDB; FungiDB:ATCC64974_10190; -.
DR VEuPathDB; FungiDB:M747DRAFT_282415; -.
DR eggNOG; KOG0157; Eukaryota.
DR BioCyc; MetaCyc:MON-15438; -.
DR BRENDA; 1.14.14.92; 518.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0043231; C:intracellular membrane-bounded organelle; IDA:UniProtKB.
DR GO; GO:0018664; F:benzoate 4-monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0020037; F:heme binding; IEA:InterPro.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR Gene3D; 1.10.630.10; -; 1.
DR InterPro; IPR001128; Cyt_P450.
DR InterPro; IPR017972; Cyt_P450_CS.
DR InterPro; IPR002401; Cyt_P450_E_grp-I.
DR InterPro; IPR036396; Cyt_P450_sf.
DR Pfam; PF00067; p450; 1.
DR PRINTS; PR00463; EP450I.
DR PRINTS; PR00385; P450.
DR SUPFAM; SSF48264; SSF48264; 1.
DR PROSITE; PS00086; CYTOCHROME_P450; 1.
PE 1: Evidence at protein level;
KW Glycoprotein; Heme; Iron; Membrane; Metal-binding; Monooxygenase;
KW Oxidoreductase; Transmembrane; Transmembrane helix.
FT CHAIN 1..517
FT /note="Benzoate 4-monooxygenase bphA"
FT /id="PRO_0000052038"
FT TRANSMEM 4..24
FT /note="Helical"
FT /evidence="ECO:0000255"
FT BINDING 461
FT /ligand="heme"
FT /ligand_id="ChEBI:CHEBI:30413"
FT /ligand_part="Fe"
FT /ligand_part_id="ChEBI:CHEBI:18248"
FT /note="axial binding residue"
FT /evidence="ECO:0000250|UniProtKB:P04798"
FT CARBOHYD 282
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 325
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
SQ SEQUENCE 517 AA; 58016 MW; 63FFBFDF703EC30B CRC64;
MLALLLSPYG AYLGLALLVL YYLLPYLKRA HLRDIPAPGL AAFTNFWLLL QTRRGHRFVV
VDNAHKKYGK LVRIAPRHTS IADDGAIQAV YGHGNGFLKS DFYDAFVSIH RGLFNTRDRA
EHTRKRKTVS HTFSMKSIGQ FEQYIHGNIE LFVKQWNRMA DTQRNPKTGF ASLDALNWFN
YLAFDIIGDL AFGAPFGMLD KGKDFAEMRK TPDSPPSYVQ AVEVLNRRGE VSATLGCYPA
LKPFAKYLPD SFFRDGIQAV EDLAGIAVAR VNERLRPEVM ANNTRVDLLA RLMEGKDSNG
EKLGRAELTA EALTQLIAGS DTTSNTSCAI LYWCMRTPGV IEKLHKALDE AIPQDVDVPT
HAMVKDIPYL QWVIWETMRI HSTSAMGLPR EIPAGNPPVT ISGHTFYPGD VVSVPSYTIH
RSKEIWGPDA EQFVPERWDP ARLTPRQKAA FIPFSTGPRA CVGRNVAEME LLVICGTVFR
LFEFEMQQEG PMETREGFLR KPLGLQVGMK RRQPGSA
