| IED ID | IndEnz0016000067 |
| Enzyme Type ID | tyrosinase000067 |
| Protein Name |
Phenylalanine ammonia-lyase hkm12 PAL EC 4.3.1.24 Hancockiamides biosynthesis cluster protein 12 |
| Gene Name | |
| Organism | Aspergillus hancockii |
| Taxonomic Lineage | cellular organisms Eukaryota Opisthokonta Fungi Dikarya Ascomycota saccharomyceta Pezizomycotina leotiomyceta Eurotiomycetes Eurotiomycetidae Eurotiales (green and blue molds) Aspergillaceae Aspergillus Aspergillus hancockii |
| Enzyme Sequence | MLSKYQPSAHIAVTRAHWEDLHQAISSGKVTIDGNSLTLADVVAVSKFGCYARLSENRETIDAINESVSTLQECLDEGHHIYGVNTGFGGSADSRTDHLASLQRALLQLLQSGILTKADIGSGDTPSQSHAMPPEWVKAIMVVRSNSVARGHSAVSIGSIEAILRLLQRDITPVVPLRGTISASGDLMPLAYIVGAIEGNPGVFARAGKSPHGQALPAQQVLEQLGIPRITLGPKEALGLVNGTAASAALSSLVLYEAHRLALLSQVTTALTVEALRGSAESFHPFISQARPHDGQMEAASNILTVMRGSRLAMGTSEVQTGLVQDRYSLRTASQWIGPQLEDLLLADRQITVELNSTTDNPLIDSVSRHFYCGGNFQATSVTSAMEKTRLALQMLGKLMFAQCSEMIDPSLNNGLPTNLVADDPSLSFTMKGVDISMAAYMSELAYLANPVSSHVQTAEMHNQAVNSLAFVSARYTMQAVDIVSMMSACSLYVACQALDLRVLQLNFFRELHPIVCNGTHDAFHTILAPKELERITQQLVTAIQDAWLTTSRMDAGDRCQRVIKLSLPILLNEMRGAIPSDRQQVDLLTSIGNWEEATCYKMLEAYKQTHERFCRTQNTVEYLGAGSKAIYHAIRHKVGVPFHQGFVEQPSADDLDTTAIINGREKKTTGGWISLIYEALRDDSLTGVILEAVQPVRSI |
| Enzyme Length | 700 |
| Uniprot Accession Number | P0DUM0 |
| Absorption | |
| Active Site | ACT_SITE 82; /note=Proton donor/acceptor; /evidence=ECO:0000250|UniProtKB:Q68G84 |
| Activity Regulation | |
| Binding Site | BINDING 242; /note=Substrate; /evidence=ECO:0000250|UniProtKB:Q68G84; BINDING 325; /note=Substrate; /evidence=ECO:0000250|UniProtKB:Q68G84; BINDING 331; /note=Substrate; /evidence=ECO:0000250|UniProtKB:Q68G84; BINDING 361; /note=Substrate; /evidence=ECO:0000250|UniProtKB:Q68G84; BINDING 463; /note=Substrate; /evidence=ECO:0000250|UniProtKB:Q68G84 |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=L-phenylalanine = (E)-cinnamate + NH4(+); Xref=Rhea:RHEA:21384, ChEBI:CHEBI:15669, ChEBI:CHEBI:28938, ChEBI:CHEBI:58095; EC=4.3.1.24; Evidence={ECO:0000255|RuleBase:RU003955};PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:21385; Evidence={ECO:0000305|PubMed:33242032}; |
| DNA Binding | |
| EC Number | 4.3.1.24 |
| Enzyme Function | FUNCTION: Phenylalanine ammonia-lyase; part of the gene cluster that mediates the biosynthesis of hancockiamides, an unusual new family of N-cinnamoylated piperazines (PubMed:33242032). The NRPS hkm10 and the NmrA-like reductase hkm9 are proposed to convert two molecules of L-Phe to the intermediary piperazine called xenocockiamide A (Probable). Xenocockiamide A is then converted to hancockiamide D via a series of hydroxylations and O-methylations (Probable). The tyrosinase hkm6 may catalyze an aromatic hydroxylation, then the 2-oxoglutarate-dependent Fe(II) dioxygenase hkm4 and the FAD-dependent phenol hydroxylase hkm7 may catalyze consecutive hydroxylations to install 2 more hydroxy groups, and the methyltransferase hkm8 probably catalyzes two methylations using 2 molecules of S-adenosyl-L-methionine (SAM) (Probable). The NRPS hkm11 activates and transfers trans-cinnamate supplied by the PAL hkm12 to hancockiamide D and produces hancockiamide A (PubMed:33242032). NRPS Hkm11 has the flexibility to tolerate the bulky hancockiamide G as a substrate and the absence of the acetyl-transferase hkm3 opens up the opportunity for hkm11 to introduce a second N-cinnamoyl moiety (PubMed:33242032). The cytochrome P450 monooxygenase hkm5 catalyzes the methylenedioxy bridge formation, converting hancockiamide A into hancockiamide G (PubMed:33242032). Hkm5 can also convert hancockiamide B into hancockiamide C, and hancockiamide D into hancockiamide H (PubMed:33242032). The N-acetyltransferase hkm3 finally transfers an acetyl group to 1-N of piperazine, converting hancockiamide A into hancockiamide B and hancockiamide G into hancockiamide C (PubMed:33242032). {ECO:0000269|PubMed:33242032, ECO:0000305|PubMed:33242032}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | PATHWAY: Secondary metabolite biosynthesis. {ECO:0000305|PubMed:33242032}. |
| nucleotide Binding | |
| Features | Active site (1); Binding site (5); Chain (1); Cross-link (1); Modified residue (1) |
| Keywords | Lyase;Phenylalanine catabolism |
| Interact With | |
| Induction | |
| Subcellular Location | |
| Modified Residue | MOD_RES 184; /note="2,3-didehydroalanine (Ser)"; /evidence="ECO:0000255|PROSITE-ProRule:PRU10122" |
| Post Translational Modification | PTM: Contains an active site 4-methylidene-imidazol-5-one (MIO), which is formed autocatalytically by cyclization and dehydration of residues Ala-Ser-Gly. {ECO:0000250|UniProtKB:Q68G84}. |
| Signal Peptide | |
| Structure 3D | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 75,940 |
| Kinetics | |
| Metal Binding | |
| Rhea ID | RHEA:21384; RHEA:21385 |
| Cross Reference Brenda |