HCHL_PSEFL
ID HCHL_PSEFL Reviewed; 276 AA.
AC O69762;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1998, sequence version 1.
DT 03-AUG-2022, entry version 90.
DE RecName: Full=Hydroxycinnamoyl-CoA hydratase-lyase;
DE Short=HCHL;
DE EC=4.1.2.61 {ECO:0000269|PubMed:10222033, ECO:0000269|PubMed:9461612};
DE AltName: Full=P-hydroxycinnamoyl CoA hydratase/lyase;
DE AltName: Full=Trans-feruloyl-CoA hydratase/vanillin synthase;
OS Pseudomonas fluorescens.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=294;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-21, FUNCTION, AND
RP CATALYTIC ACTIVITY.
RC STRAIN=AN103;
RX PubMed=9461612; DOI=10.1074/jbc.273.7.4163;
RA Gasson M.J., Kitamura Y., McLauchlan W.R., Narbad A., Parr A.J.,
RA Parsons E.L., Payne J., Rhodes M.J., Walton N.J.;
RT "Metabolism of ferulic acid to vanillin. A bacterial gene of the enoyl-SCoA
RT hydratase/isomerase superfamily encodes an enzyme for the hydration and
RT cleavage of a hydroxycinnamic acid SCoA thioester.";
RL J. Biol. Chem. 273:4163-4170(1998).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=AN103;
RX PubMed=10222033; DOI=10.1006/abbi.1999.1140;
RA Mitra A., Kitamura Y., Gasson M.J., Narbad A., Parr A.J., Payne J.,
RA Rhodes M.J., Sewter C., Walton N.J.;
RT "4-hydroxycinnamoyl-CoA hydratase/lyase (HCHL)--An enzyme of
RT phenylpropanoid chain cleavage from Pseudomonas.";
RL Arch. Biochem. Biophys. 365:10-16(1999).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS), AND SUBUNIT.
RC STRAIN=AN103;
RX PubMed=17139085; DOI=10.1107/s0907444906039199;
RA Leonard P.M., Brzozowski A.M., Lebedev A., Marshall C.M., Smith D.J.,
RA Verma C.S., Walton N.J., Grogan G.;
RT "The 1.8 A resolution structure of hydroxycinnamoyl-coenzyme A hydratase-
RT lyase (HCHL) from Pseudomonas fluorescens, an enzyme that catalyses the
RT transformation of feruloyl-coenzyme A to vanillin.";
RL Acta Crystallogr. D 62:1494-1501(2006).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH
RP 4-HYDROXY-3-METHOXYBENZALDEHYDE AND ACETYL-COA, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBUNIT, AND MUTAGENESIS OF SER-123; GLU-143 AND TYR-239.
RC STRAIN=AN103;
RX PubMed=18479250; DOI=10.1042/bj20080714;
RA Bennett J.P., Bertin L., Moulton B., Fairlamb I.J., Brzozowski A.M.,
RA Walton N.J., Grogan G.;
RT "A ternary complex of hydroxycinnamoyl-CoA hydratase-lyase (HCHL) with
RT acetyl-CoA and vanillin gives insights into substrate specificity and
RT mechanism.";
RL Biochem. J. 414:281-289(2008).
CC -!- FUNCTION: Catalyzes the hydration of the acyl-CoA thioester of ferulic
CC acid and the subsequent retro-aldol cleavage of the hydrated
CC intermediate to yield vanillin (4-hydroxy-3-methoxy-benzaldehyde)
CC (PubMed:9461612, PubMed:10222033). The enzyme is also active with
CC caffeoyl-CoA and 4-coumaroyl-CoA producing 3,4-dihydroxybenzaldehyde
CC and 4-hydroxybenaldehyde, respectively (PubMed:10222033).
CC {ECO:0000269|PubMed:10222033, ECO:0000269|PubMed:9461612}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(E)-caffeoyl-CoA + H2O = 3,4-dihydroxybenzaldehyde + acetyl-
CC CoA; Xref=Rhea:RHEA:36307, ChEBI:CHEBI:15377, ChEBI:CHEBI:50205,
CC ChEBI:CHEBI:57288, ChEBI:CHEBI:87136; EC=4.1.2.61;
CC Evidence={ECO:0000269|PubMed:10222033};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(E)-feruloyl-CoA + H2O = acetyl-CoA + vanillin;
CC Xref=Rhea:RHEA:62412, ChEBI:CHEBI:15377, ChEBI:CHEBI:18346,
CC ChEBI:CHEBI:57288, ChEBI:CHEBI:87305; EC=4.1.2.61;
CC Evidence={ECO:0000269|PubMed:10222033, ECO:0000269|PubMed:9461612};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(E)-4-coumaroyl-CoA + H2O = 4-hydroxybenzaldehyde + acetyl-
CC CoA; Xref=Rhea:RHEA:62416, ChEBI:CHEBI:15377, ChEBI:CHEBI:17597,
CC ChEBI:CHEBI:57288, ChEBI:CHEBI:85008; EC=4.1.2.61;
CC Evidence={ECO:0000269|PubMed:10222033};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=11.8 uM for feruloyl-CoA {ECO:0000269|PubMed:18479250};
CC KM=2.4 uM for feruloyl-CoA {ECO:0000269|PubMed:10222033};
CC KM=5.3 uM for 4-coumaroyl-CoA {ECO:0000269|PubMed:10222033};
CC KM=1.6 uM for caffeoyl-CoA {ECO:0000269|PubMed:10222033};
CC Vmax=36.5 nmol/sec/mg enzyme with feruloyl-CoA as substrate
CC {ECO:0000269|PubMed:10222033};
CC Vmax=73.3 nmol/sec/mg enzyme with 4-coumaroyl-CoA as substrate
CC {ECO:0000269|PubMed:10222033};
CC Vmax=15.2 nmol/sec/mg enzyme with caffeoyl-CoA as substrate
CC {ECO:0000269|PubMed:10222033};
CC Note=kcat is 3.72 sec(-1) with feruloyl-CoA as substrate.
CC {ECO:0000269|PubMed:18479250};
CC pH dependence:
CC Optimum pH is 8.5-9.5. {ECO:0000269|PubMed:10222033};
CC -!- SUBUNIT: Homohexamer; dimer of trimers. {ECO:0000269|PubMed:17139085,
CC ECO:0000269|PubMed:18479250}.
CC -!- SIMILARITY: Belongs to the enoyl-CoA hydratase/isomerase family.
CC {ECO:0000305}.
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DR EMBL; Y13067; CAA73502.1; -; Genomic_DNA.
DR RefSeq; WP_027620477.1; NZ_CP078138.1.
DR PDB; 2J5I; X-ray; 1.80 A; A/B/C/D/E/F/G/H/I/J/K/L=1-276.
DR PDB; 2VSS; X-ray; 2.22 A; A/B/C/D/E/F=1-276.
DR PDB; 2VSU; X-ray; 1.90 A; A/B/C/D/E/F=1-276.
DR PDBsum; 2J5I; -.
DR PDBsum; 2VSS; -.
DR PDBsum; 2VSU; -.
DR AlphaFoldDB; O69762; -.
DR SMR; O69762; -.
DR STRING; 690597.JH730982_gene2032; -.
DR eggNOG; COG1024; Bacteria.
DR BRENDA; 4.1.2.61; 5121.
DR EvolutionaryTrace; O69762; -.
DR GO; GO:0050547; F:vanillin synthase activity; IEA:UniProtKB-EC.
DR InterPro; IPR029045; ClpP/crotonase-like_dom_sf.
DR InterPro; IPR018376; Enoyl-CoA_hyd/isom_CS.
DR InterPro; IPR001753; Enoyl-CoA_hydra/iso.
DR Pfam; PF00378; ECH_1; 1.
DR SUPFAM; SSF52096; SSF52096; 1.
DR PROSITE; PS00166; ENOYL_COA_HYDRATASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Lyase.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:9461612"
FT CHAIN 2..276
FT /note="Hydroxycinnamoyl-CoA hydratase-lyase"
FT /id="PRO_0000418727"
FT BINDING 29
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT BINDING 70..72
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT BINDING 75
FT /ligand="substrate"
FT BINDING 120
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT BINDING 142..146
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT BINDING 151
FT /ligand="substrate"
FT BINDING 239
FT /ligand="substrate"
FT MUTAGEN 123
FT /note="S->A: Reduced kcat compared to wild-type but not
FT markerdly."
FT /evidence="ECO:0000269|PubMed:18479250"
FT MUTAGEN 143
FT /note="E->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:18479250"
FT MUTAGEN 239
FT /note="Y->F: Increased KM but retains a significant amount
FT of catalytic activity with a kcat 10 times less than that
FT of the wild-type."
FT /evidence="ECO:0000269|PubMed:18479250"
FT CONFLICT 16
FT /note="E -> Q (in Ref. 1; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 9..16
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 18..24
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 27..29
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 35..49
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 54..62
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 65..67
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 72..81
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 86..98
FT /evidence="ECO:0007829|PDB:2J5I"
FT TURN 99..104
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 109..113
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 115..118
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 119..121
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 122..127
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 128..134
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 138..140
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 142..146
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 154..161
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 164..173
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 176..178
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 179..184
FT /evidence="ECO:0007829|PDB:2J5I"
FT STRAND 187..192
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 194..209
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 213..226
FT /evidence="ECO:0007829|PDB:2J5I"
FT HELIX 231..248
FT /evidence="ECO:0007829|PDB:2J5I"
SQ SEQUENCE 276 AA; 31008 MW; FDB5AD1539CA7F9A CRC64;
MSTYEGRWKT VKVEIEDGIA FVILNRPEKR NAMSPTLNRE MIDVLETLEQ DPAAGVLVLT
GAGEAWTAGM DLKEYFREVD AGPEILQEKI RREASQWQWK LLRMYAKPTI AMVNGWCFGG
GFSPLVACDL AICADEATFG LSEINWGIPP GNLVSKAMAD TVGHRQSLYY IMTGKTFGGQ
KAAEMGLVNE SVPLAQLREV TIELARNLLE KNPVVLRAAK HGFKRCRELT WEQNEDYLYA
KLDQSRLLDT EGGREQGMKQ FLDDKSIKPG LQAYKR
