PHNH_PENHR
ID PHNH_PENHR Reviewed; 149 AA.
AC A0A1S6PUA4;
DT 22-APR-2020, integrated into UniProtKB/Swiss-Prot.
DT 10-MAY-2017, sequence version 1.
DT 25-MAY-2022, entry version 14.
DE RecName: Full=Hydroalkoxylation enzyme phnH {ECO:0000303|PubMed:26978228};
DE EC=4.-.-.- {ECO:0000269|PubMed:28240554};
DE AltName: Full=Phenalenone biosynthesis cluster protein H {ECO:0000303|PubMed:26978228};
DE Flags: Precursor;
GN Name=phnH {ECO:0000303|PubMed:28240554};
OS Penicillium herquei.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Penicillium.
OX NCBI_TaxID=69774;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RC STRAIN=ATCC 10118 / CBS 336.48 / NBRC 31747 / NRRL 1040;
RX PubMed=28240554; DOI=10.1021/jacs.7b01089;
RA Gao S.S., Garcia-Borras M., Barber J.S., Hai Y., Duan A., Garg N.K.,
RA Houk K.N., Tang Y.;
RT "Enzyme-catalyzed intramolecular enantioselective hydroalkoxylation.";
RL J. Am. Chem. Soc. 139:3639-3642(2017).
RN [2]
RP FUNCTION.
RX PubMed=26978228; DOI=10.1021/jacs.6b01528;
RA Gao S.S., Duan A., Xu W., Yu P., Hang L., Houk K.N., Tang Y.;
RT "Phenalenone polyketide cyclization catalyzed by fungal polyketide synthase
RT and flavin-dependent monooxygenase.";
RL J. Am. Chem. Soc. 138:4249-4259(2016).
CC -!- FUNCTION: Hydroalkoxylation enzyme; part of the gene cluster that
CC mediates the biosynthesis of phenalenones such as herqueinone,
CC compounds that have been reported to treat tumors, bacterial infections
CC and/or mycoses, and rheumatic diseases (PubMed:26978228). The non-
CC reducing polyketide synthase phnA synthesizes the heptaketide backbone
CC and cyclizes it into the angular, hemiketal-containing naphtho-gamma-
CC pyrone prephenalenone. The product template (PT) domain of phnA
CC catalyzes only the C4-C9 aldol condensation, which is unprecedented
CC among known PT domains (PubMed:28240554, PubMed:26978228). The
CC transformation of prephenalenone to phenalenones requires an FAD-
CC dependent monooxygenase phnB, which catalyzes the C2 aromatic
CC hydroxylation of prephenalenone and ring opening of the gamma-pyrone
CC ring simultaneously (PubMed:28240554, PubMed:26978228). Subsequent
CC intramolecular deprotonation of C3 phenolic oxygen accelerates
CC phenalenone ring closure to yield the tricyclic phenalenone core with a
CC C2 hydroxylation (PubMed:28240554, PubMed:26978228). The
CC prenyltransferase phnF further catalyzes reverse C-prenylation of
CC phenalenone by direct electrophilic substitution at C6, or possibly via
CC first a forward O-prenylation of a neighboring phenol in phenalenone,
CC followed by a Claisen rearrangement (PubMed:28240554). The
CC hydroalkoxylation enzyme phnH catalyzes the 5-exo-trigcyclization via
CC acid catalysis after the spontaneous deprotonation of 7-OH, which leads
CC to the formation of the dihydrobenzofuran atrovenetin
CC (PubMed:28240554). Atrovenetin is further converted to deoxyherqueinone
CC by the O-methyltransferase phnC which can methylate C2-OH to stabilize
CC the northern portion of the phenalenone core (PubMed:28240554).
CC Finally, the oxidoreductase phnG converts deoxyherqueinone to
CC herqueinone via C6 hydroxylation (PubMed:28240554).
CC {ECO:0000269|PubMed:26978228, ECO:0000269|PubMed:28240554}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2,4,7,9-tetrahydroxy-6-methyl-8-(2-methylbut-3-en-2-yl)-1-oxo-
CC 1H-phenalen-3-ol = (2'R)-atrovenetin; Xref=Rhea:RHEA:62660,
CC ChEBI:CHEBI:145870, ChEBI:CHEBI:145872;
CC Evidence={ECO:0000269|PubMed:28240554};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62661;
CC Evidence={ECO:0000269|PubMed:28240554};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=216 uM for the atrovenetin precursor
CC {ECO:0000269|PubMed:28240554};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:28240554};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:28240554}.
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DR EMBL; KY475538; AQV03780.1; -; mRNA.
DR PDB; 6JJS; X-ray; 1.62 A; A=1-149.
DR PDB; 6JJT; X-ray; 1.33 A; A/B/C/D=1-149.
DR PDBsum; 6JJS; -.
DR PDBsum; 6JJT; -.
DR AlphaFoldDB; A0A1S6PUA4; -.
DR SMR; A0A1S6PUA4; -.
DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW.
PE 1: Evidence at protein level;
KW 3D-structure; Glycoprotein; Lyase; Signal.
FT SIGNAL 1..18
FT /evidence="ECO:0000255"
FT CHAIN 19..149
FT /note="Hydroalkoxylation enzyme phnH"
FT /id="PRO_5013249856"
FT CARBOHYD 33
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT CARBOHYD 127
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT STRAND 25..36
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 40..45
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 48..61
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 66..79
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 84..94
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 99..107
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 110..116
FT /evidence="ECO:0007829|PDB:6JJT"
FT TURN 120..122
FT /evidence="ECO:0007829|PDB:6JJT"
FT HELIX 123..127
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 130..133
FT /evidence="ECO:0007829|PDB:6JJT"
FT STRAND 142..149
FT /evidence="ECO:0007829|PDB:6JJT"
SQ SEQUENCE 149 AA; 16038 MW; 51CE55ACB1D04179 CRC64;
MKFTYLVSLA AFAVTALGSR PTPPNLEFLF SANLTKGPAY IYDQSDAQIK ALQTLTGGII
AGPNFDGTVI GGTALSTRGA DGTIRADAHY LIQTSDGANI LVTESAAIPY VAVLFDTSSE
KYNWLNNVTA WGTPPNLNEI NFLEYWQIE
