{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T06:43:10Z","timestamp":1787640190322,"version":"build-2736575974"},"reference-count":42,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2024,4,29]],"date-time":"2024-04-29T00:00:00Z","timestamp":1714348800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2021YFC2100600"],"award-info":[{"award-number":["2021YFC2100600"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2023YFC3402400"],"award-info":[{"award-number":["2023YFC3402400"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2024,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Avermectins (AVEs), a family of macrocyclic polyketides produced by\n                    <jats:italic>Streptomyces avermitilis<\/jats:italic>\n                    , have eight components, among which B1a is noted for its strong insecticidal activity. Biosynthesis of AVE \u201ca\u201d components requires 2\u2010methylbutyryl\u2010CoA (MBCoA) as starter unit, and malonyl\u2010CoA (MalCoA) and methylmalonyl\u2010CoA (MMCoA) as extender units. We describe here a novel strategy for increasing B1a production by enhancing acyl\u2010CoA precursor supply. First, we engineered meilingmycin (MEI) polyketide synthase (PKS) for increasing MBCoA precursor supply. The loading module (using acetyl\u2010CoA as substrate), extension module 7 (using MMCoA as substrate) and TE domain of MEI PKS were assembled to produce 2\u2010methylbutyrate, providing the starter unit for B1a production. Heterologous expression of the newly designed PKS (termed Mei\u2010PKS) in\n                    <jats:italic>S.\u2009avermitilis<\/jats:italic>\n                    wild\u2010type (WT) strain increased MBCoA level, leading to B1a titer 262.2\u2009\u03bcg\/mL \u2013 4.36\u2010fold higher than WT value (48.9\u2009\u03bcg\/mL). Next, we separately inhibited three key nodes in essential pathways using CRISPRi to increase MalCoA and MMCoA levels in WT. The resulting strains all showed increased B1a titer. Combined inhibition of these key nodes in Mei\u2010PKS expression strain increased B1a titer to 341.9\u2009\u03bcg\/mL. Overexpression of fatty acid \u03b2\u2010oxidation pathway genes in the strain further increased B1a titer to 452.8\u2009\u03bcg\/mL \u2013 8.25\u2010fold higher than WT value. Finally, we applied our precursor supply strategies to high\u2010yield industrial strain A229. The strategies, in combination, led to B1a titer 8836.4\u2009\u03bcg\/mL \u2013 37.8% higher than parental A229 value. These findings provide an effective combination strategy for increasing AVE B1a production in WT and industrial\n                    <jats:italic>S.\u2009avermitilis<\/jats:italic>\n                    strains, and our precursor supply strategies can be readily adapted for overproduction of other polyketides.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.14470","type":"journal-article","created":{"date-parts":[[2024,4,29]],"date-time":"2024-04-29T11:54:00Z","timestamp":1714391640000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Enhancement of acyl\u2010\n                    <scp>CoA<\/scp>\n                    precursor supply for increased avermectin B1a production by engineering meilingmycin polyketide synthase and key primary metabolic pathway genes"],"prefix":"10.1111","volume":"17","author":[{"given":"Mengyao","family":"Yang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Animal Biotech Breeding and College of Biological Sciences China Agricultural University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Hao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Animal Biotech Breeding and College of Biological Sciences China Agricultural University  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3583-2985","authenticated-orcid":false,"given":"Gang","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Mycology Institute of Microbiology, Chinese Academy of Sciences  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8455-6900","authenticated-orcid":false,"given":"Ying","family":"Wen","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Animal Biotech Breeding and College of Biological Sciences China Agricultural 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