HKM10_ASPHA
ID HKM10_ASPHA Reviewed; 1020 AA.
AC P0DUL8;
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=Non-canonical nonribosomal peptide synthetase hkm10 {ECO:0000303|PubMed:33242032};
DE EC=6.3.2.- {ECO:0000305|PubMed:33242032};
DE AltName: Full=Hancockiamides biosynthesis cluster protein 10 {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: Non-canonical nonribosomal peptide synthetase; part of the
CC gene cluster that mediates the biosynthesis of hancockiamides, an
CC unusual new family of N-cinnamoylated piperazines (PubMed:33242032).
CC The NRPS hkm10 and the NmrA-like reductase hkm9 are proposed to convert
CC two molecules of L-Phe to the intermediary piperazine called
CC xenocockiamide A (Probable). Xenocockiamide A is then converted to
CC hancockiamide D via a series of hydroxylations and O-methylations
CC (Probable). The tyrosinase hkm6 may catalyze an aromatic hydroxylation,
CC then the 2-oxoglutarate-dependent Fe(II) dioxygenase hkm4 and the FAD-
CC dependent phenol hydroxylase hkm7 may catalyze consecutive
CC hydroxylations to install 2 more hydroxy groups, and the
CC methyltransferase hkm8 probably catalyzes two methylations using 2
CC molecules of S-adenosyl-L-methionine (SAM) (Probable). The NRPS hkm11
CC activates and transfers trans-cinnamate supplied by the PAL hkm12 to
CC hancockiamide D and produces hancockiamide A (PubMed:33242032). NRPS
CC Hkm11 has the flexibility to tolerate the bulky hancockiamide G as a
CC substrate and the absence of the acetyl-transferase hkm3 opens up the
CC opportunity for hkm11 to introduce a second N-cinnamoyl moiety
CC (PubMed:33242032). The cytochrome P450 monooxygenase hkm5 catalyzes the
CC methylenedioxy bridge formation, converting hancockiamide A into
CC hancockiamide G (PubMed:33242032). Hkm5 can also convert hancockiamide
CC B into hancockiamide C, and hancockiamide D into hancockiamide H
CC (PubMed:33242032). The N-acetyltransferase hkm3 finally transfers an
CC acetyl group to 1-N of piperazine, converting hancockiamide A into
CC hancockiamide B and hancockiamide G into hancockiamide C
CC (PubMed:33242032). {ECO:0000269|PubMed:33242032,
CC ECO:0000305|PubMed:33242032}.
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000305|PubMed:33242032}.
CC -!- DOMAIN: NRP synthetases are composed of discrete domains (adenylation
CC (A), thiolation (T) or peptidyl carrier protein (PCP) and condensation
CC (C) domains) which when grouped together are referred to as a single
CC module. Each module is responsible for the recognition (via the A
CC domain) and incorporation of a single amino acid into the growing
CC peptide product. Thus, an NRP synthetase is generally composed of one
CC or more modules and can terminate in a thioesterase domain (TE) that
CC releases the newly synthesized peptide from the enzyme. Occasionally,
CC epimerase (E) domains (responsible for L- to D-amino acid conversion)
CC are present within the NRP synthetase. Hkm10 contains an amino acid
CC adenylation domain (A), a peptidyl carrier protein (PCP) domain with a
CC phosphopantetheine prosthetic group, and a short-chain
CC dehydrogenase/reductase terminus (R), but it does not have an
CC identifiable condensation (C) domain required for the formation of
CC peptide bonds during non-ribosomal peptide synthesis.
CC {ECO:0000305|PubMed:33242032}.
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 NRP synthetase family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; MBFL02000005; KAF7597140.1; -; Genomic_DNA.
DR AlphaFoldDB; P0DUL8; -.
DR SMR; P0DUL8; -.
DR GO; GO:0016874; F:ligase activity; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0009058; P:biosynthetic process; IEA:UniProt.
DR CDD; cd05235; SDR_e1; 1.
DR Gene3D; 1.10.1200.10; -; 1.
DR Gene3D; 3.30.300.30; -; 1.
DR Gene3D; 3.40.50.12780; -; 1.
DR InterPro; IPR036736; ACP-like_sf.
DR InterPro; IPR045851; AMP-bd_C_sf.
DR InterPro; IPR020845; AMP-binding_CS.
DR InterPro; IPR000873; AMP-dep_Synth/Lig.
DR InterPro; IPR042099; ANL_N_sf.
DR InterPro; IPR013120; Far_NAD-bd.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR009081; PP-bd_ACP.
DR InterPro; IPR010080; Thioester_reductase-like_dom.
DR Pfam; PF00501; AMP-binding; 1.
DR Pfam; PF07993; NAD_binding_4; 1.
DR Pfam; PF00550; PP-binding; 1.
DR SUPFAM; SSF47336; SSF47336; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR01746; Thioester-redct; 1.
DR PROSITE; PS00455; AMP_BINDING; 1.
DR PROSITE; PS50075; CARRIER; 1.
PE 1: Evidence at protein level;
KW Ligase; Multifunctional enzyme; Oxidoreductase; Phosphopantetheine;
KW Phosphoprotein.
FT CHAIN 1..1020
FT /note="Non-canonical nonribosomal peptide synthetase hkm10"
FT /id="PRO_0000452937"
FT DOMAIN 526..608
FT /note="Carrier"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258,
FT ECO:0000305|PubMed:33242032"
FT REGION 21..419
FT /note="Adenylation (A) domain"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:33242032"
FT REGION 652..894
FT /note="short-chain dehydrogenase/reductase (R) domain"
FT /evidence="ECO:0000255, ECO:0000305|PubMed:33242032"
FT MOD_RES 568
FT /note="O-(pantetheine 4'-phosphoryl)serine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00258"
SQ SEQUENCE 1020 AA; 111844 MW; 630C3715C5AC64B7 CRC64;
MSTFESLLAR NEGLGQLFYQ QMLEDPDAIA IVDGDYSLTY ASLHAQATHL AQRLDQNDFV
HEEPVGIVVQ HGILDAVAQV AIIYAGGTCV TLDPALPNQQ IERRLNRLRA RYILVDTPNK
SRGLPFSQIE VEDLPISTEL IPTDSPYPVN LSLEHRSHLI HTSGTTSESK AVQIVGRSIV
HVANYAPFEP VVKTDVVAHG NSTSFDVALF DIWAPLVQGA SIAVLSKATL LDLSAFEAAI
DRYKISVMAI TAPLVNLAAT TRPGMFSSMR VVLMGGEAVN IPAMRKIFEA GPPVHMVNAY
GPTECCVYCL ARKITLEDLD TGAVSIGKAI GNNIATVCDE MGKPVPDGEE GELLVGGPGV
SPGYVNLPGK NAASFIEVPD LVDANGTPYH MYRTGDLVKR RPDGQYDFVG RFDHQVKIRG
YRVELGAIET VLMDTGYFSE GVVMKVDSKA EGAGSALVAF AVLAPTAPPS AVTDATAALT
AALPHYMIPN IHIVESIPLT NHAKVDRKQL ADWCLQRQEK NMCAMQDKVP SEGASTRDQL
GALWATILAT PVREYSDNDD FFGLGGTSLQ ASLLISLIRR TFNTEVSLLA LYDNSTLGQL
AHIVDRNQGG ALATVQNLRE MWIADTMIGD ALETPVGPVV DWRRDTEGRV FLTGATGFVG
AFLLSDMLKM PGIHQVGCLV RAPDEATGVR RLRHALEKYN LWREEYLPKL LPLCGKLEDP
WLGLGEQRFR EIADWASVIF HLGALVNYTQ PYSWHRPANI EGTVNVVRLA CTGRSKALHY
CSSISCFGPT GIINGTKVVH EDGALMPHLN ALPYDHGYAQ SQWVAEELLR RLIHRRFPIA
VYRPGFITGH SETGACNPDD FFSRLIRACS SIGCYPGLPN QRKEFVPIDY VTSTMIHIAS
SSLSLGHAFH IVPPTREESP EMNDTMSLIG ELTGTSIQPV SYREWIEQLS STKDLSLQPL
LPMLAEVVID GMTRWEMYEN MPTYENTNTL RALASCPDLP KFPMVDEALL RKYLDYLADH
