{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T17:44:30Z","timestamp":1786729470314,"version":"build-2736575974"},"reference-count":44,"publisher":"American Chemical Society (ACS)","issue":"8","license":[{"start":{"date-parts":[[2022,7,13]],"date-time":"2022-07-13T00:00:00Z","timestamp":1657670400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2022,7,13]],"date-time":"2022-07-13T00:00:00Z","timestamp":1657670400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2022,7,13]],"date-time":"2022-07-13T00:00:00Z","timestamp":1657670400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100002855","name":"Ministry of Science and Technology of the People's Republic of China","doi-asserted-by":"publisher","award":["2021YFC2102100"],"award-info":[{"award-number":["2021YFC2102100"]}],"id":[{"id":"10.13039\/501100002855","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Program of Shanghai Technology Research Leader","award":["19XD1433200"],"award-info":[{"award-number":["19XD1433200"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,8,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The compound FR901379, a sulfated echinocandin produced by the filamentous fungus Coleophoma empetri F-11899, is an important intermediate for the synthesis of the antifungal drug micafungin. In this study, we established an efficient clustered regularly interspaced short palindromic repeats\/Cas9-based gene editing tool for the industrial production strain C. empetri SIPI1284. With this method, the efficiency of gene mutagenesis in the target locus is up to 84%, which enables the rapid gene disruption for the analysis of FR901379 biosynthetic genes. Next, we verified the putative functional genes of the FR901379 biosynthetic gene cluster via gene disruption and gene complementation in vivo. These core functional genes included the nonribosomal peptide synthetase gene (CEnrps), the fatty-acyl-AMP ligase gene (CEligase) responsible for the formation of the activated form of palmitic acid and its transfer to CEnrps, four nonheme mononuclear iron oxygenase genes (CEoxy1, CEoxy2, CEoxy3, and CEoxy4) responsible for the synthesis of nonproteinogenic amino acids, l-homotyrosine biosynthesis genes (CEhtyA-D), two cytochrome P450 enzyme genes (CEp450-1 and CEp450-2), and a transcription regulator gene (CEhyp). In addition, by screening the whole genome, we identified two unknown genes (CEp450-3 and CEsul) responsible for the sulfonyloxy group of FR901379, which were separated from the core FR901379 biosynthetic cluster. Furthermore, during gene disruptions in the research, we obtained a series of FR901379 analogues and elucidated the relationship between the groups and antifungal activities.<\/jats:p>","DOI":"10.1021\/acschembio.2c00250","type":"journal-article","created":{"date-parts":[[2022,7,13]],"date-time":"2022-07-13T03:52:55Z","timestamp":1657684375000},"page":"2130-2141","source":"Crossref","is-referenced-by-count":10,"title":["Analysis of FR901379\nBiosynthetic Genes in\n                    <i>Coleophoma empetri<\/i>\n                    by Clustered Regularly Interspaced\nShort Palindromic Repeats\/Cas9-Based Genomic Manipulation"],"prefix":"10.1021","volume":"17","author":[{"given":"Teng-Yun","family":"Wei","sequence":"first","affiliation":[{"name":"Shanghai Institute of Pharmaceutical Industry , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Zheng","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Pharmaceutical Industry , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miyang","family":"Wan","sequence":"additional","affiliation":[{"name":"Fudan University , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Songbai","family":"Yang","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Pharmaceutical Industry , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiawei","family":"Tang","sequence":"additional","affiliation":[{"name":"Fudan University , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanjie","family":"Wu","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Pharmaceutical Industry , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyang","family":"Li","sequence":"additional","affiliation":[{"name":"Fudan University , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1604-816X","authenticated-orcid":true,"given":"Shao-Xin","family":"Chen","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Pharmaceutical Industry , , ,","place":["Shanghai, China, 201203"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2022,7,13]]},"reference":[{"key":"2026081413090828900_cit1","doi-asserted-by":"publisher","first-page":"1092","DOI":"10.7164\/antibiotics.47.1092","article-title":"WF11899A, B and C, Novel Antifungal\nLipopetides","volume":"47","author":"Iwamoto","year":"1994","journal-title":"J. Antibiot."},{"key":"2026081413090828900_cit2","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1038\/ja.2008.3","article-title":"Micafungin:\na sulfated echinocandin","volume":"62","author":"Hashimoto","year":"2009","journal-title":"J. Antibiot."},{"key":"2026081413090828900_cit3","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1351\/pac200779040603","article-title":"Discovery\nof micafungin (FK463): A novel antifungal drug derived from a natural\nproduct lead","volume":"79","author":"Fujie","year":"2007","journal-title":"Pure Appl. Chem."},{"key":"2026081413090828900_cit4","doi-asserted-by":"publisher","first-page":"16781","DOI":"10.1021\/ja307220z","article-title":"Identification\nand Characterization of the Echinocandin B Biosynthetic Gene Cluster\nfrom Emericella rugulosa NRRL 11440","volume":"134","author":"Cacho","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"2026081413090828900_cit5","doi-asserted-by":"publisher","first-page":"2724","DOI":"10.1021\/acschembio.6b00604","article-title":"Engineering of New\nPneumocandin Side-Chain Analogues from Glarea lozoyensis by Mutasynthesis and Evaluation of Their Antifungal Activity","volume":"11","author":"Chen","year":"2016","journal-title":"ACS Chem. Biol."},{"key":"2026081413090828900_cit6","doi-asserted-by":"publisher","first-page":"1550","DOI":"10.1128\/aem.03256-14","article-title":"Engineering\nof Glarea lozoyensis for Exclusive\nProduction of the Pneumocandin B0 Precursor of the Antifungal\nDrug Caspofungin Acetate","volume":"81","author":"Chen","year":"2015","journal-title":"Appl. Environ. Microbiol."},{"key":"2026081413090828900_cit7","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1186\/1471-2164-14-339","article-title":"Genomics-driven discovery\nof the pneumocandin biosynthetic gene cluster\nin the fungus Glarea lozoyensis","volume":"14","author":"Chen","year":"2013","journal-title":"BMC Genomics"},{"key":"2026081413090828900_cit8","doi-asserted-by":"publisher","first-page":"4457","DOI":"10.1021\/ja312572v","article-title":"EcdGHK are three\ntailoring iron oxygenases for amino\nacid building blocks of the echinocandin scaffold","volume":"135","author":"Jiang","year":"2013","journal-title":"J. Am. Chem. Soc."},{"key":"2026081413090828900_cit9","doi-asserted-by":"publisher","first-page":"1702","DOI":"10.1021\/acschembio.5b00013","article-title":"Genetic Manipulation of the Pneumocandin\nBiosynthetic\nPathway for Generation of Analogues and Evaluation of Their Antifungal\nActivity","volume":"10","author":"Li","year":"2015","journal-title":"ACS Chem. Biol."},{"key":"2026081413090828900_cit10","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.02370-17","article-title":"Cryptic Production\nof trans-3-Hydroxyproline in Echinocandin B Biosynthesis","volume":"84","author":"Mattay","year":"2018","journal-title":"Appl. Environ. Microbiol."},{"key":"2026081413090828900_cit11","doi-asserted-by":"publisher","first-page":"7877","DOI":"10.1007\/s00253-018-9179-y","article-title":"Biochemical and genetic characterization of fungal\nproline hydroxylase in echinocandin biosynthesis","volume":"102","author":"Zhang","year":"2018","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"2026081413090828900_cit12","doi-asserted-by":"publisher","first-page":"623","DOI":"10.1007\/s00294-009-0275-5","article-title":"Agrobacterium tumefaciens-mediated transformation\nof antifungal lipopeptide producing fungus Coleophoma\nempetri F-11899","volume":"55","author":"Yamada","year":"2009","journal-title":"Curr. Genet."},{"key":"2026081413090828900_cit13","doi-asserted-by":"publisher","first-page":"734780","DOI":"10.3389\/fmicb.2021.734780","article-title":"Establishing an efficient Genetic Manipulation System\nfor Sulfated\nEchinocandin Producing Fungus Coleophoma empetri","volume":"12","author":"Men","year":"2021","journal-title":"Front. Microbiol."},{"key":"2026081413090828900_cit14","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.1500248","article-title":"A Candida albicans CRISPR system\npermits genetic engineering of essential genes and gene families","volume":"1","author":"Vyas","year":"2015","journal-title":"Sci. Adv."},{"key":"2026081413090828900_cit15","doi-asserted-by":"publisher","first-page":"1134","DOI":"10.1021\/acssynbio.9b00024","article-title":"A CRISPR-Cas9\nStrategy for Activating the Saccharopolyspora erythraea Erythromycin Biosynthetic Gene Cluster with Knock-in Bidirectional\nPromoters","volume":"8","author":"Liu","year":"2019","journal-title":"ACS Synth. Biol."},{"key":"2026081413090828900_cit16","doi-asserted-by":"publisher","first-page":"15007","DOI":"10.1038\/celldisc.2015.7","article-title":"Efficient\ngenome editing in filamentous fungus Trichoderma reesei using the CRISPR\/Cas9 system","volume":"1","author":"Liu","year":"2015","journal-title":"Cell Discovery"},{"key":"2026081413090828900_cit17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/ncomms6344","article-title":"Implementation\nof the CRISPR-Cas9 system in fission yeast","volume":"5","author":"Jacobs","year":"2015","journal-title":"Nat. Commun."},{"key":"2026081413090828900_cit18","doi-asserted-by":"publisher","first-page":"1262","DOI":"10.1016\/j.cell.2014.05.010","article-title":"Development\nand Applications of CRISPR-Cas9\nfor Genome Engineering","volume":"157","author":"Hsu","year":"2014","journal-title":"Cell"},{"key":"2026081413090828900_cit19","doi-asserted-by":"publisher","first-page":"910","DOI":"10.1016\/j.cell.2013.04.025","article-title":"One-Step Generation\nof Mice Carrying\nMutations in Multiple Genes by CRISPR\/Cas-Mediated Genome Engineering","volume":"153","author":"Wang","year":"2013","journal-title":"Cell"},{"key":"2026081413090828900_cit20","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex Genome Engineering\nUsing CRISPR\/Cas\nSystems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"2026081413090828900_cit21","doi-asserted-by":"publisher","first-page":"1568","DOI":"10.1021\/acssynbio.7b00456","article-title":"5S rRNA promoter for guide RNA expression enabled highly\nefficient CRISPR\/Cas9 genome editing in Aspergillus\nniger","volume":"8","author":"Zheng","year":"2019","journal-title":"ACS Synth. Biol."},{"key":"2026081413090828900_cit22","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.fgb.2015.12.007","article-title":"Highly efficient CRISPR\nmutagenesis by microhomology-mediated end joining in Aspergillus fumigatus","volume":"86","author":"Zhang","year":"2016","journal-title":"Fungal\nGenet. Biol."},{"key":"2026081413090828900_cit23","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1021\/acssynbio.8b00478","article-title":"CRISPR\/Cas9-Based\nGenome Editing in the Filamentous Fungus Fusarium fujikuroi and Its Application in Strain\nEngineering for Gibberellic Acid Production","volume":"8","author":"Shi","year":"2019","journal-title":"ACS Synth. Biol."},{"key":"2026081413090828900_cit24","doi-asserted-by":"publisher","first-page":"754","DOI":"10.1021\/acssynbio.6b00082","article-title":"CRISPR\/Cas9\nBased Genome Editing\nof Penicillium chrysogenum","volume":"5","author":"Pohl","year":"2016","journal-title":"ACS Synth. 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Biofuels"},{"key":"2026081413090828900_cit26","doi-asserted-by":"publisher","first-page":"637","DOI":"10.1007\/s10529-015-2015-x","article-title":"Development\nof a genome editing technique\nusing the CRISPR\/Cas9 system in the industrial filamentous fungus Aspergillus oryzae","volume":"38","author":"Katayama","year":"2016","journal-title":"Biotechnol.\nLett."},{"key":"2026081413090828900_cit27","doi-asserted-by":"publisher","first-page":"1968","DOI":"10.1021\/acssynbio.9b00491","article-title":"CRISPR\/Cas9-Based\nGenome Editing in the Filamentous Fungus Glarea lozoyensis and Its Application in Manipulating gloF","volume":"9","author":"Wei","year":"2020","journal-title":"ACS Synth. Biol."},{"key":"2026081413090828900_cit28","doi-asserted-by":"publisher","first-page":"645","DOI":"10.1007\/s00438-002-0780-4","article-title":"Efficient disruption of a polyketide\nsynthase gene (pks1) required for melanin synthesis\nthrough Agrobacterium-mediated transformation\nof Glarea lozoyensis","volume":"268","author":"Zhang","year":"2003","journal-title":"Mol. Genet. Genomics"},{"key":"2026081413090828900_cit29","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1186\/s12896-015-0165-5","article-title":"Development of genetic\ntools for Myceliophthora thermophila","volume":"15","author":"Xu","year":"2015","journal-title":"BMC Biotechnol."},{"key":"2026081413090828900_cit30","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41467-022-28501-7","article-title":"CRISPR-assisted\nrational flux-tuning and arrayed CRISPRi screening of an L-proline\nexporter for L-proline hyperproduction","volume":"13","author":"Liu","year":"2022","journal-title":"Nat.\nCommun."},{"key":"2026081413090828900_cit31","doi-asserted-by":"publisher","first-page":"3031","DOI":"10.1128\/jb.180.12.3031-3038.1998","article-title":"The developmentally\nregulated alb1 gene of Aspergillus fumigatus: its role in modulation of conidial morphology and virulence","volume":"180","author":"Tsai","year":"1998","journal-title":"J. Bacteriol."},{"key":"2026081413090828900_cit32","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1007\/bf00290362","article-title":"Structural analysis of PKS1, a polyketide synthase\ngene involved in melanin biosynthesis in Colletotrichum\nlagenarium","volume":"249","author":"Takano","year":"1995","journal-title":"Mol. Gen. Genet."},{"key":"2026081413090828900_cit33","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1007\/bf00279362","article-title":"The developmentally regulated Aspergillus\nnidulanswA gene encodes a polypeptide\nhomologous to polyketide and fatty acid synthases","volume":"235","author":"Mayorga","year":"1992","journal-title":"Mol. Gen. Genet."},{"key":"2026081413090828900_cit34","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1007\/s004300050077","article-title":"Identification of\na polyketide synthase gene (pksP) of Aspergillus fumigatus involved in conidial pigment\nbiosynthesis and virulence","volume":"187","author":"Langfelder","year":"1998","journal-title":"Med. Microbiol.\nImmunol."},{"key":"2026081413090828900_cit35","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1007\/s004380051133","article-title":"A melanin polyketide synthase (PKS)\ngene from Nodulisporium sp. that shows\nhomology to the pks1 gene of Colletotrichum\nlagenarium","volume":"262","author":"Fulton","year":"1999","journal-title":"Mol. Gen. Genet."},{"key":"2026081413090828900_cit36","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1038\/nchembio.1047","article-title":"Cyclization of Fungal Nonribosomal\nPeptides by a Terminal Condensation-Like Domain","volume":"8","author":"Gao","year":"2012","journal-title":"Nat. Chem. Biol."},{"key":"2026081413090828900_cit37","doi-asserted-by":"publisher","first-page":"3770","DOI":"10.1002\/anie.201609079","article-title":"Nonribosomal Peptide\nSynthesis\ue5f8Principles and Prospects","volume":"56","author":"Su\u0308ssmuth","year":"2017","journal-title":"Angew.\nChem."},{"key":"2026081413090828900_cit38","doi-asserted-by":"publisher","first-page":"3468","DOI":"10.1021\/cr0503097","article-title":"Assembly-Line Enzymology\nfor Polyketide and Nonribosomal\nPeptide Antibiotics: Logic, Machinery, and Mechanisms","volume":"106","author":"Fischbach","year":"2006","journal-title":"Chem. Rev."},{"key":"2026081413090828900_cit39","doi-asserted-by":"publisher","first-page":"698","DOI":"10.1128\/ec.00076-15","article-title":"Evolution\nof Chemical Diversity in\nEchinocandin Lipopeptide Antifungal Metabolites","volume":"14","author":"Yue","year":"2015","journal-title":"Eukaryotic Cell"},{"key":"2026081413090828900_cit40","doi-asserted-by":"publisher","first-page":"1247","DOI":"10.1089\/ars.2010.3119","article-title":"The PAPS-independent\naryl sulfotransferase and the\nalternative disulfide bond formation system in pathogenic bacteria","volume":"13","author":"Malojcic","year":"2010","journal-title":"Antioxid. Redox Signaling"},{"key":"2026081413090828900_cit41","doi-asserted-by":"publisher","first-page":"1385","DOI":"10.1126\/science.1243106","article-title":"Genetic and molecular basis of drug resistance and\nspecies-specific drug action in schistosome parasites","volume":"342","author":"Valentim","year":"2013","journal-title":"Science"},{"key":"2026081413090828900_cit42","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1038\/87617","article-title":"A helical lid converts\na sulfotransferase to a dehydratase","volume":"8","author":"Pakhomova","year":"2001","journal-title":"Nat. Struct.\nBiol."},{"key":"2026081413090828900_cit43","doi-asserted-by":"publisher","first-page":"1994","DOI":"10.1021\/cb300385m","article-title":"Structural Basis\nof Functional Group Activation by Sulfotransferases in Complex Metabolic\nPathways","volume":"7","author":"McCarthy","year":"2012","journal-title":"ACS Chem. Biol."},{"key":"2026081413090828900_cit44","doi-asserted-by":"publisher","first-page":"18724","DOI":"10.1073\/pnas.0806975105","article-title":"Redirecting the substrate specificity\nof heparan sulfate 2-O-sulfotransferase by structurally guided mutagenesis","volume":"105","author":"Bethea","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U.S.A."}],"container-title":["ACS Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acschembio.2c00250","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/17\/8\/2130\/3731588\/cb2c00250.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/17\/8\/2130\/3731588\/cb2c00250.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T17:24:16Z","timestamp":1786728256000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acbcct\/article\/17\/8\/2130\/384775\/Analysis-of-FR901379-Biosynthetic-Genes-in"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,13]]},"references-count":44,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,7,13]]},"published-print":{"date-parts":[[2022,8,19]]}},"URL":"https:\/\/doi.org\/10.1021\/acschembio.2c00250","relation":{},"ISSN":["1554-8929","1554-8937"],"issn-type":[{"value":"1554-8929","type":"print"},{"value":"1554-8937","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,13]]}}}