GLOH_GLAL2
ID GLOH_GLAL2 Reviewed; 617 AA.
AC S3D9F8;
DT 20-JUN-2018, integrated into UniProtKB/Swiss-Prot.
DT 18-SEP-2013, sequence version 1.
DT 25-MAY-2022, entry version 31.
DE RecName: Full=Isopropyl malate synthase gloH {ECO:0000250|UniProtKB:K0E4E5};
DE EC=2.3.3.13 {ECO:0000250|UniProtKB:K0E4E5};
DE AltName: Full=L-homotyrosine biosynthesis sub-cluster protein gloH {ECO:0000303|PubMed:23688303};
DE AltName: Full=Pneumocandin biosynthesis cluster protein H {ECO:0000303|PubMed:25270390};
GN Name=gloH {ECO:0000303|PubMed:25270390};
GN Synonyms=GLHtyA {ECO:0000303|PubMed:25879325}; ORFNames=GLAREA_10039;
OS Glarea lozoyensis (strain ATCC 20868 / MF5171).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Leotiomycetes;
OC Helotiales; Helotiaceae; Glarea.
OX NCBI_TaxID=1116229;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA], IDENTIFICATION, AND
RP FUNCTION.
RC STRAIN=ATCC 20868 / MF5171;
RX PubMed=23688303; DOI=10.1186/1471-2164-14-339;
RA Chen L., Yue Q., Zhang X., Xiang M., Wang C., Li S., Che Y.,
RA Ortiz-Lopez F.J., Bills G.F., Liu X., An Z.;
RT "Genomics-driven discovery of the pneumocandin biosynthetic gene cluster in
RT the fungus Glarea lozoyensis.";
RL BMC Genomics 14:339-339(2013).
RN [2]
RP FUNCTION.
RX PubMed=25270390; DOI=10.1002/cbic.201402175;
RA Houwaart S., Youssar L., Huettel W.;
RT "Pneumocandin biosynthesis: involvement of a trans-selective proline
RT hydroxylase.";
RL ChemBioChem 15:2365-2369(2014).
RN [3]
RP FUNCTION, PATHWAY, AND BIOTECHNOLOGY.
RX PubMed=25879325; DOI=10.1021/acschembio.5b00013;
RA Li Y., Chen L., Yue Q., Liu X., An Z., Bills G.F.;
RT "Genetic manipulation of the pneumocandin biosynthetic pathway for
RT generation of analogues and evaluation of their antifungal activity.";
RL ACS Chem. Biol. 10:1702-1710(2015).
RN [4]
RP FUNCTION, AND BIOTECHNOLOGY.
RX PubMed=25527531; DOI=10.1128/aem.03256-14;
RA Chen L., Yue Q., Li Y., Niu X., Xiang M., Wang W., Bills G.F., Liu X.,
RA An Z.;
RT "Engineering of Glarea lozoyensis for exclusive production of the
RT pneumocandin B0 precursor of the antifungal drug caspofungin acetate.";
RL Appl. Environ. Microbiol. 81:1550-1558(2015).
RN [5]
RP FUNCTION, AND BIOTECHNOLOGY.
RX PubMed=27494047; DOI=10.1021/acschembio.6b00604;
RA Chen L., Li Y., Yue Q., Loksztejn A., Yokoyama K., Felix E.A., Liu X.,
RA Zhang N., An Z., Bills G.F.;
RT "Engineering of new pneumocandin side-chain analogues from Glarea
RT lozoyensis by mutasynthesis and evaluation of their antifungal activity.";
RL ACS Chem. Biol. 11:2724-2733(2016).
RN [6]
RP FUNCTION.
RX PubMed=29352089; DOI=10.1128/aem.02370-17;
RA Mattay J., Houwaart S., Huettel W.;
RT "Cryptic production of trans-3-hydroxyproline in echinocandin B
RT biosynthesis.";
RL Appl. Environ. Microbiol. 0:0-0(2018).
RN [7]
RP REVIEW.
RX PubMed=27705900; DOI=10.1515/znc-2016-0156;
RA Huettel W.;
RT "Structural diversity in echinocandin biosynthesis: the impact of oxidation
RT steps and approaches toward an evolutionary explanation.";
RL Z. Naturforsch. C 72:1-20(2017).
CC -!- FUNCTION: 2-isopropylmalate synthase; part of the gene cluster that
CC mediates the biosynthesis of pneumocandins, lipohexapeptides of the
CC echinocandin family that prevent fungal cell wall formation by non-
CC competitive inhibition of beta-1,3-glucan synthase (PubMed:27705900).
CC The 10,12-dimethylmyristoyl side chain is synthesized by the reducing
CC polyketide synthase gloL/GLPKS4 (PubMed:27494047). The thioesterase
CC gloN/GLHYD exclusively interacts with gloL/GLPKS4 to maintain turnover
CC of the polyketide side chain (PubMed:27494047). The 10R,12S-
CC dimethylmyristic acid is then transferred to the first thiolation
CC domain of the nonribosomal peptide synthetase gloA/GLNRPS4 by the acyl-
CC AMP ligase gloD/GLligase, followed by its acylation to L-ornithine to
CC trigger elongation of the cyclic hexapeptide (PubMed:27494047). L-
CC ornithine, 4R-hydroxyl-L-proline (generated from L-proline by the
CC dioxygenase gloF/GLOXY2), 3S-hydroxyl-L-homotyrosine (generated by
CC gloG/GLHtyB, gloH/GLHtyA, gloI/GLHtyC, gloJ/GLHtyD and hydroxylated at
CC C-3 by the dioxygenase gloM/GLOXY1), 3R-hydroxyl-L-glutamine (generated
CC from L-glutamine probably by the dioxygenase gloE/GLOXY3) and 3S-
CC hydroxyl-L-proline (generated from L-proline by the dioxygenase
CC gloF/GLOXY2 to yield pneumocandin B0), or 3S-hydroxyl-4S-methyl-L-
CC proline (generated from L-leucine by the dioxygenase gloC/GLOXY4 to
CC yield pneumocandin A0) are sequentially added to the growing chain
CC (PubMed:25270390, PubMed:25879325, PubMed:25527531). The last C domain
CC of gloA/GLNRPS4 is proposed to be responsible for cyclization by
CC condensation to form the peptide bond between L-ornithine and 3S-
CC hydroxyl-4S-methyl-L-proline (for pneumocandin A0) or 3S-hydroxyl-L-
CC proline (for pneumocandin B0). Finally, the subsequent C-4
CC hydroxylation of 3S-hydroxyl-L-homotyrosine and L-ornithine
CC dihydroxylation at C-4 and C-5 are performed by the cytochrome P450
CC monooxygenases gloP/GLP450-1 and gloO/GLP450-2, respectively
CC (PubMed:25879325). {ECO:0000269|PubMed:25270390,
CC ECO:0000269|PubMed:25527531, ECO:0000269|PubMed:25879325,
CC ECO:0000269|PubMed:27494047, ECO:0000269|PubMed:29352089,
CC ECO:0000303|PubMed:27705900}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-methyl-2-oxobutanoate + acetyl-CoA + H2O = (2S)-2-
CC isopropylmalate + CoA + H(+); Xref=Rhea:RHEA:21524, ChEBI:CHEBI:1178,
CC ChEBI:CHEBI:11851, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57287, ChEBI:CHEBI:57288; EC=2.3.3.13;
CC Evidence={ECO:0000250|UniProtKB:K0E4E5};
CC -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000305|PubMed:25879325}.
CC -!- BIOTECHNOLOGY: Pneumocandin B0 is the starting molecule for the first
CC semisynthetic echinocandin antifungal drug, caspofungin acetate
CC (PubMed:25527531). Pneumocandin B0 is a minor fermentation product, and
CC its industrial production was achieved by a combination of extensive
CC mutation and medium optimization (PubMed:25527531). Inactivation of
CC three of gloP/GLP450-1, gloO/GLP450-2, and gloM/GLOXY1 generates 13
CC different pneumocandin analogs that lack one, two, three, or four
CC hydroxyl groups on 4R,5R-dihydroxy-ornithine and 3S,4S-dihydroxy-
CC homotyrosine of the parent hexapeptide (PubMed:25879325). All of these
CC cyclic lipopeptides show potent antifungal activities, and two new
CC metabolites pneumocandins F and G are more potent in vitro against
CC Candida species and Aspergillus fumigatus than the principal
CC fermentation products, pneumocandins A0 and B0 (PubMed:25879325).
CC Moreover, feeding alternative side chain precursors yields acrophiarin
CC and 4 additional pneumocandin congeners with straight C14, C15, and C16
CC side chains. One of those compounds, pneumocandin I, has elevated
CC antifungal activity and similar hemolytic activity compared to
CC pneumocandin B0, the starting molecule for caspofungin, demonstrating
CC the potential for using gloD/GLligase for future engineering of new
CC echinocandin analogs (PubMed:27494047). {ECO:0000269|PubMed:25527531,
CC ECO:0000269|PubMed:25879325, ECO:0000269|PubMed:27494047}.
CC -!- SIMILARITY: Belongs to the alpha-IPM synthase/homocitrate synthase
CC family. LeuA type 2 subfamily. {ECO:0000305}.
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DR EMBL; KE145356; EPE34345.1; -; Genomic_DNA.
DR RefSeq; XP_008078280.1; XM_008080089.1.
DR AlphaFoldDB; S3D9F8; -.
DR SMR; S3D9F8; -.
DR STRING; 1116229.S3D9F8; -.
DR EnsemblFungi; EPE34345; EPE34345; GLAREA_10039.
DR GeneID; 19469086; -.
DR KEGG; glz:GLAREA_10039; -.
DR eggNOG; KOG2367; Eukaryota.
DR HOGENOM; CLU_004588_3_0_1; -.
DR OrthoDB; 882838at2759; -.
DR Proteomes; UP000016922; Unassembled WGS sequence.
DR GO; GO:0003852; F:2-isopropylmalate synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0009098; P:leucine biosynthetic process; IEA:UniProtKB-KW.
DR Gene3D; 3.20.20.70; -; 1.
DR Gene3D; 3.30.160.270; -; 1.
DR InterPro; IPR002034; AIPM/Hcit_synth_CS.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR036230; LeuA_allosteric_dom_sf.
DR InterPro; IPR000891; PYR_CT.
DR Pfam; PF00682; HMGL-like; 1.
DR PROSITE; PS00815; AIPM_HOMOCIT_SYNTH_1; 1.
DR PROSITE; PS00816; AIPM_HOMOCIT_SYNTH_2; 1.
DR PROSITE; PS50991; PYR_CT; 1.
PE 1: Evidence at protein level;
KW Amino-acid biosynthesis; Branched-chain amino acid biosynthesis;
KW Leucine biosynthesis; Reference proteome; Transferase.
FT CHAIN 1..617
FT /note="Isopropyl malate synthase gloH"
FT /id="PRO_0000444495"
FT DOMAIN 47..325
FT /note="Pyruvate carboxyltransferase"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01151"
SQ SEQUENCE 617 AA; 68088 MW; B53774CCFF138F67 CRC64;
MKNTDERYSM ATCCAGGQSL RSKYGERAPN IQLSDRQWPS KKLTKSPIWL STDLRDGNQA
LPNPMTLQQK WRMFQLLVSI GFTQIEVSFP CASQTEFDFT RQLIETPGAT PDDVTLEVLS
PCREDAIATS VRSLKGAKQA ILFLYLATSD NYRETVLQLS EAEWLDQAKR CVEYARSITK
DDPENCRTKW SFGFGFEDFA NARPEAALRL GETIKTAWGP SKDNPVILGL ATSVEATTPN
VFADQVEYFS RNITEREKVC ISIHTHNDRG GAVASAELAC LAGGDRVEGC LFGNGERAGN
LDLITCSMNM FTQGIETGLD FSDLLEIRRV YEEVTNLPVH PRQPYSGDFY FRAFAGAHQD
AIRKGLAKRG RLAAMPSGLT QMSKGMIING TSNEKIPSST WRVPYLPLDP ADLGFSFDSV
IGVNSQSGKG GIAWLIQQGL GFNIPNELAA HYSQIIKKRS VSLERGLAAE EICDFFLEIY
DLKSSYNTQL LKRLHSGFSQ TIDMQQELED ATAPANNAAM DISKTMGIDV HCTMASSHVI
SSHDGEQLSN VAFVQCEIEQ TTPCVWGVGV ALSREEAVNR SLLLAARRKP GTETPDTKTS
VNAIIQPLEV YSFSQKA
