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 16-JAN-2019, entry version 22. 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}; GN 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 RT cluster in 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 RT 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 RT 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 RT oxidation 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 CC that mediates the biosynthesis of pneumocandins, lipohexapeptides CC of the echinocandin family that prevent fungal cell wall formation CC by non-competitive inhibition of beta-1,3-glucan synthase CC (PubMed:27705900). The 10,12-dimethylmyristoyl side chain is CC synthesized by the reducing polyketide synthase gloL/GLPKS4 CC (PubMed:27494047). The thioesterase gloN/GLHYD exclusively CC interacts with gloL/GLPKS4 to maintain turnover of the polyketide CC side chain (PubMed:27494047). The 10R,12S-dimethylmyristic acid is CC then transferred to the first thiolation domain of the CC nonribosomal peptide synthetase gloA/GLNRPS4 by the acyl-AMP CC 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 CC hydroxylated at C-3 by the dioxygenase gloM/GLOXY1), 3R-hydroxyl- CC L-glutamine (generated from L-glutamine probably by the CC dioxygenase gloE/GLOXY3) and 3S-hydroxyl-L-proline (generated from CC L-proline by the dioxygenase gloF/GLOXY2 to yield pneumocandin CC B0), or 3S-hydroxyl-4S-methyl-L-proline (generated from L-leucine CC by the dioxygenase gloC/GLOXY4 to yield pneumocandin A0) are CC sequentially added to the growing chain (PubMed:25270390, CC PubMed:25879325, PubMed:25527531). The last C domain of CC 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- CC L-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 CC P450 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 CC analogs with non-hydroxylated ornithine (PubMed:25879325). CC {ECO:0000269|PubMed:25879325}. CC -!- BIOTECHNOLOGY: Pneumocandin B0 is the starting molecule for the CC first semisynthetic echinocandin antifungal drug, caspofungin CC acetate (PubMed:25527531). Pneumocandin B0 is a minor fermentation CC product, and its industrial production was achieved by a CC combination of extensive mutation and medium optimization CC (PubMed:25527531). Inactivation of three of gloP/GLP450-1, CC gloO/GLP450-2, and gloM/GLOXY1 generates 13 different pneumocandin CC analogs that lack one, two, three, or four hydroxyl groups on CC 4R,5R-dihydroxy-ornithine and 3S,4S-dihydroxy-homotyrosine of the CC parent hexapeptide (PubMed:25879325). All of these cyclic CC 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 CC acrophiarin and 4 additional pneumocandin congeners with straight CC C14, C15, and C16 side chains. One of those compounds, CC pneumocandin I, has elevated antifungal activity and similar CC hemolytic activity compared to pneumocandin B0, the starting CC molecule for caspofungin, demonstrating the potential for using CC gloD/GLligase for future engineering of new echinocandin analogs CC (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 EnsemblFungi; EPE34337; EPE34337; GLAREA_10031. DR GeneID; 19469078; -. DR KEGG; glz:GLAREA_10031; -. 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 Complete proteome; Heme; Iron; Metal-binding; Monooxygenase; KW Oxidoreductase; Reference proteome; Signal. FT SIGNAL 1 26 {ECO:0000255}. FT CHAIN 27 521 Cytochrome P450 monooxygenase gloO. FT {ECO:0000255}. FT /FTId=PRO_0000444489. FT METAL 464 464 Iron (heme axial ligand). FT {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 //