Evolution-guided engineering of trans-acyltransferase polyketide synthases

• 成果类型: Article
• 署名作者: Mabesoone, Mathijs F. J.; Leopold-Messer, Stefan; Minas, Hannah A.; Chepkirui, Clara; Chawengrum, Pornsuda; Reiter, Silke; Meoded, Roy A.; Wolf, Sarah; Genz, Ferdinand; Magnus, Nancy; Piechulla, Birgit; Walker, Allison S.; Piel, Jorn
• 署名单位: Chulabhorn Royal Academy; Chulabhorn Graduate Institute; University of Rostock; Harvard University; Harvard Medical School; Vanderbilt University; Vanderbilt University
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-10608
• DOI: 10.1126/science.adj7621
• 发表日期: 2024-03-22
• 页码: 1312-1317
• 关键词: biosynthetic pathways; assembly lines; product; ketosynthase; bacillaene; sequence; complex; genome; domain; identification

摘要

Bacterial multimodular polyketide synthases (PKSs) are giant enzymes that generate a wide range of therapeutically important but synthetically challenging natural products. Diversification of polyketide structures can be achieved by engineering these enzymes. However, notwithstanding successes made with textbook cis-acyltransferase (cis-AT) PKSs, tailoring such large assembly lines remains challenging. Unlike textbook PKSs, trans-AT PKSs feature an extraordinary diversity of PKS modules and commonly evolve to form hybrid PKSs. In this study, we analyzed amino acid coevolution to identify a common module site that yields functional PKSs. We used this site to insert and delete diverse PKS parts and create 22 engineered trans-AT PKSs from various pathways and in two bacterial producers. The high success rates of our engineering approach highlight the broader applicability to generate complex designer polyketides.