ID GLOO_GLAL2 Reviewed; 521 AA. AC S3D775; DT 20-JUN-2018, integrated into UniProtKB/Swiss-Prot. DT 18-SEP-2013, sequence version 1. DT 25-MAY-2022, entry version 28. DE RecName: Full=Cytochrome P450 monooxygenase gloO {ECO:0000303|PubMed:25270390}; DE EC=1.-.-.- {ECO:0000305|PubMed:25879325}; DE AltName: Full=Ornithine 3,4-hydroxylase {ECO:0000303|PubMed:27705900}; DE AltName: Full=Pneumocandin biosynthesis cluster protein O {ECO:0000303|PubMed:25270390}; DE Flags: Precursor; GN Name=gloO {ECO:0000303|PubMed:25270390}; GN Synonyms=GLP450-2 {ECO:0000303|PubMed:25879325}; ORFNames=GLAREA_10031; 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, DISRUPTION PHENOTYPE, 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: Cytochrome P450 monooxygenase; 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 -!- COFACTOR: CC Name=heme; Xref=ChEBI:CHEBI:30413; CC Evidence={ECO:0000250|UniProtKB:P04798}; CC -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000269|PubMed:25879325}. CC -!- DISRUPTION PHENOTYPE: Leads to the production of pneumocandin analogs CC with non-hydroxylated ornithine (PubMed:25879325). CC {ECO:0000269|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 cytochrome P450 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; KE145356; EPE34337.1; -; Genomic_DNA. DR RefSeq; XP_008078272.1; XM_008080081.1. DR AlphaFoldDB; S3D775; -. DR SMR; S3D775; -. DR EnsemblFungi; EPE34337; EPE34337; GLAREA_10031. DR GeneID; 19469078; -. DR KEGG; glz:GLAREA_10031; -. DR eggNOG; KOG0156; Eukaryota. DR HOGENOM; CLU_022195_9_2_1; -. DR OrthoDB; 595327at2759; -. DR Proteomes; UP000016922; Unassembled WGS sequence. DR GO; GO:0020037; F:heme binding; IEA:InterPro. DR GO; GO:0005506; F:iron ion binding; IEA:InterPro. DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW. DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro. DR Gene3D; 1.10.630.10; -; 1. DR InterPro; IPR001128; Cyt_P450. DR InterPro; IPR017972; Cyt_P450_CS. DR InterPro; IPR002403; Cyt_P450_E_grp-IV. DR InterPro; IPR036396; Cyt_P450_sf. DR Pfam; PF00067; p450; 1. DR PRINTS; PR00465; EP450IV. DR SUPFAM; SSF48264; SSF48264; 1. DR PROSITE; PS00086; CYTOCHROME_P450; 1. PE 1: Evidence at protein level; KW Heme; Iron; Metal-binding; Monooxygenase; Oxidoreductase; KW Reference proteome; Signal. FT SIGNAL 1..26 FT /evidence="ECO:0000255" FT CHAIN 27..521 FT /note="Cytochrome P450 monooxygenase gloO" FT /evidence="ECO:0000255" FT /id="PRO_0000444489" FT METAL 464 FT /note="Iron (heme axial ligand)" FT /evidence="ECO:0000250|UniProtKB:P04798" SQ SEQUENCE 521 AA; 58099 MW; BE0523C57DA14944 CRC64; MIAALFTTNL QLGAVGVFIF ALLAFAFNKL TTWEYSIPKE VQWVDRRTQP FSYLRAKARA LARSKENTLE AYFRFNKLGK AAALAVPFGR PLLLLPQTFV RWIVDQPESI ISLDPIHDDF HVFVGGDLTG DHTVQELLRR ELTLNLDKLI SVINDEIVCA LDDVLGNSPE WKSTSLADDL KTIVARTSNR VFVGKDLCRN KHYISTVKGL ALVIMPETVL QDLIPQFLKG PLSRITKAFN NIYGMKKFSS LLLGVVRQRY IDVKDVLEGS GDKTRLPDDL LTWMVQKSIR KGESSANIDK LLVARIAMAN LAAIETTTAA MTRSVLDLVT QGSEGGFLKA VQEETLAVVE GCNYEPSKKD VLKLVLTENA IKEALRLQVA FPGLMRQVVA PNGVTLENGL HVPCGTRLGV SAAGIHVDES IYEDPTTYNP GRFLVRDLDP RGDPSPMWKG NENYLAFSLG RRSCPGRWYV TDQLKLTLAH IFSKYEIRFE KAAETASALR KILPGAPQDR VMIRRRSVGK R //