HKM12_ASPHA
ID HKM12_ASPHA Reviewed; 700 AA.
AC P0DUM0;
DT 02-JUN-2021, integrated into UniProtKB/Swiss-Prot.
DT 02-JUN-2021, sequence version 1.
DT 03-AUG-2022, entry version 5.
DE RecName: Full=Phenylalanine ammonia-lyase hkm12 {ECO:0000303|PubMed:33242032};
DE Short=PAL {ECO:0000303|PubMed:33242032};
DE EC=4.3.1.24 {ECO:0000305|PubMed:33242032};
DE AltName: Full=Hancockiamides biosynthesis cluster protein 12 {ECO:0000303|PubMed:33242032};
OS Aspergillus hancockii.
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus.
OX NCBI_TaxID=1873369;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=FRR 3425 / CBS 142004 / DTO 360-G7;
RA Gilchrist C.L.M., Chooi Y.H.;
RL Submitted (APR-2019) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, PATHWAY, AND BIOTECHNOLOGY.
RX PubMed=33242032; DOI=10.1039/d0ob02243h;
RA Li H., Lacey A.E., Shu S., Kalaitzis J.A., Vuong D., Crombie A., Hu J.,
RA Gilchrist C.L.M., Lacey E., Piggott A.M., Chooi Y.H.;
RT "Hancockiamides: phenylpropanoid piperazines from Aspergillus hancockii are
RT biosynthesised by a versatile dual single-module NRPS pathway.";
RL Org. Biomol. Chem. 19:587-595(2021).
CC -!- FUNCTION: Phenylalanine ammonia-lyase; part of the gene cluster that
CC mediates the biosynthesis of hancockiamides, an unusual new family of
CC N-cinnamoylated piperazines (PubMed:33242032). The NRPS hkm10 and the
CC NmrA-like reductase hkm9 are proposed to convert two molecules of L-Phe
CC to the intermediary piperazine called xenocockiamide A (Probable).
CC Xenocockiamide A is then converted to hancockiamide D via a series of
CC hydroxylations and O-methylations (Probable). The tyrosinase hkm6 may
CC catalyze an aromatic hydroxylation, then the 2-oxoglutarate-dependent
CC Fe(II) dioxygenase hkm4 and the FAD-dependent phenol hydroxylase hkm7
CC may catalyze consecutive hydroxylations to install 2 more hydroxy
CC groups, and the methyltransferase hkm8 probably catalyzes two
CC methylations using 2 molecules of S-adenosyl-L-methionine (SAM)
CC (Probable). The NRPS hkm11 activates and transfers trans-cinnamate
CC supplied by the PAL hkm12 to hancockiamide D and produces hancockiamide
CC A (PubMed:33242032). NRPS Hkm11 has the flexibility to tolerate the
CC bulky hancockiamide G as a substrate and the absence of the acetyl-
CC transferase hkm3 opens up the opportunity for hkm11 to introduce a
CC second N-cinnamoyl moiety (PubMed:33242032). The cytochrome P450
CC monooxygenase hkm5 catalyzes the methylenedioxy bridge formation,
CC converting hancockiamide A into hancockiamide G (PubMed:33242032). Hkm5
CC can also convert hancockiamide B into hancockiamide C, and
CC hancockiamide D into hancockiamide H (PubMed:33242032). The N-
CC acetyltransferase hkm3 finally transfers an acetyl group to 1-N of
CC piperazine, converting hancockiamide A into hancockiamide B and
CC hancockiamide G into hancockiamide C (PubMed:33242032).
CC {ECO:0000269|PubMed:33242032, ECO:0000305|PubMed:33242032}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-phenylalanine = (E)-cinnamate + NH4(+);
CC Xref=Rhea:RHEA:21384, ChEBI:CHEBI:15669, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:58095; EC=4.3.1.24;
CC Evidence={ECO:0000255|RuleBase:RU003955};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:21385;
CC Evidence={ECO:0000305|PubMed:33242032};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000305|PubMed:33242032}.
CC -!- PTM: Contains an active site 4-methylidene-imidazol-5-one (MIO), which
CC is formed autocatalytically by cyclization and dehydration of residues
CC Ala-Ser-Gly. {ECO:0000250|UniProtKB:Q68G84}.
CC -!- BIOTECHNOLOGY: Hancockiamide D displays potent cytotoxic activity
CC against murine myeloma NS-1 cells, suggesting a potential antitumour
CC application (PubMed:33242032). More interestingly, hancockiamide C, the
CC likely end metabolite of the hkm pathway, shows potent Arabidopsis
CC thaliana seed anti-germination activity, but is inactive against the
CC monocot Eragrostis tef seed, suggesting that it could be a herbicidal
CC lead targeting monocots (PubMed:33242032). The herbicidal activity of
CC hancockiamide C could be due to its phenylpropanoid-like structural
CC features, which may act on the plant lignan pathways, and hence
CC warrants further investigations (PubMed:33242032).
CC {ECO:0000269|PubMed:33242032}.
CC -!- SIMILARITY: Belongs to the PAL/histidase family. {ECO:0000305}.
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DR EMBL; MBFL02000005; KAF7597138.1; -; Genomic_DNA.
DR AlphaFoldDB; P0DUM0; -.
DR SMR; P0DUM0; -.
DR GO; GO:0005737; C:cytoplasm; IEA:InterPro.
DR GO; GO:0045548; F:phenylalanine ammonia-lyase activity; IEA:UniProtKB-EC.
DR GO; GO:0006559; P:L-phenylalanine catabolic process; IEA:UniProtKB-KW.
DR CDD; cd00332; PAL-HAL; 1.
DR Gene3D; 1.10.274.20; -; 1.
DR Gene3D; 1.10.275.10; -; 1.
DR InterPro; IPR001106; Aromatic_Lyase.
DR InterPro; IPR024083; Fumarase/histidase_N.
DR InterPro; IPR008948; L-Aspartase-like.
DR InterPro; IPR022313; Phe/His_NH3-lyase_AS.
DR InterPro; IPR005922; Phe_NH3-lyase.
DR InterPro; IPR023144; Phe_NH3-lyase_shielding_dom_sf.
DR PANTHER; PTHR10362; PTHR10362; 1.
DR Pfam; PF00221; Lyase_aromatic; 1.
DR SUPFAM; SSF48557; SSF48557; 1.
DR TIGRFAMs; TIGR01226; phe_am_lyase; 1.
DR PROSITE; PS00488; PAL_HISTIDASE; 1.
PE 1: Evidence at protein level;
KW Lyase; Phenylalanine catabolism.
FT CHAIN 1..700
FT /note="Phenylalanine ammonia-lyase hkm12"
FT /id="PRO_0000452939"
FT ACT_SITE 82
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
FT BINDING 242
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
FT BINDING 325
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
FT BINDING 331
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
FT BINDING 361
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
FT BINDING 463
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
FT MOD_RES 184
FT /note="2,3-didehydroalanine (Ser)"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10122"
FT CROSSLNK 183..185
FT /note="5-imidazolinone (Ala-Gly)"
FT /evidence="ECO:0000250|UniProtKB:Q68G84"
SQ SEQUENCE 700 AA; 75940 MW; FF5216B73B2C7A86 CRC64;
MLSKYQPSAH IAVTRAHWED LHQAISSGKV TIDGNSLTLA DVVAVSKFGC YARLSENRET
IDAINESVST LQECLDEGHH IYGVNTGFGG SADSRTDHLA SLQRALLQLL QSGILTKADI
GSGDTPSQSH AMPPEWVKAI MVVRSNSVAR GHSAVSIGSI EAILRLLQRD ITPVVPLRGT
ISASGDLMPL AYIVGAIEGN PGVFARAGKS PHGQALPAQQ VLEQLGIPRI TLGPKEALGL
VNGTAASAAL SSLVLYEAHR LALLSQVTTA LTVEALRGSA ESFHPFISQA RPHDGQMEAA
SNILTVMRGS RLAMGTSEVQ TGLVQDRYSL RTASQWIGPQ LEDLLLADRQ ITVELNSTTD
NPLIDSVSRH FYCGGNFQAT SVTSAMEKTR LALQMLGKLM FAQCSEMIDP SLNNGLPTNL
VADDPSLSFT MKGVDISMAA YMSELAYLAN PVSSHVQTAE MHNQAVNSLA FVSARYTMQA
VDIVSMMSAC SLYVACQALD LRVLQLNFFR ELHPIVCNGT HDAFHTILAP KELERITQQL
VTAIQDAWLT TSRMDAGDRC QRVIKLSLPI LLNEMRGAIP SDRQQVDLLT SIGNWEEATC
YKMLEAYKQT HERFCRTQNT VEYLGAGSKA IYHAIRHKVG VPFHQGFVEQ PSADDLDTTA
IINGREKKTT GGWISLIYEA LRDDSLTGVI LEAVQPVRSI
