Josemarco Mendoza-Avila, Volker Döring, Madeleine Bouzon, Ivan Dubois, Tanja Knaus, Louis M. M. Mouterde, Anne Zaparucha, Francesco G. Mutti, Carine Vergne-Vaxelaire
{"title":"通过适应性进化产生的耐胺大肠杆菌菌株用于手性胺的可持续合成","authors":"Josemarco Mendoza-Avila, Volker Döring, Madeleine Bouzon, Ivan Dubois, Tanja Knaus, Louis M. M. Mouterde, Anne Zaparucha, Francesco G. Mutti, Carine Vergne-Vaxelaire","doi":"10.1021/acssuschemeng.4c04356","DOIUrl":null,"url":null,"abstract":"The biocatalytic synthesis of chiral amines from carbonyl compounds and ammonia is a major advance in sustainable synthetic chemistry. Using whole cells for bioamination reactions is advantageous given their low preparation cost and direct applicability; however, amine toxicity limits the reaction when living cells are used. Herein, we adapted <i>Escherichia coli</i> BL21(DE3) cells to grow in the presence of 100 mM hexan-2-amine <i>via</i> directed evolution in continuous culture, obtaining six times more tolerant strains than the wild-type. The adapted strains also displayed superior tolerance for structurally different amines. Coexpression of genes encoding for amine dehydrogenase (AmDH) and formate dehydrogenase activities in the adapted strains enabled the stereoselective bioamination (ee > 99%) of different prochiral ketones with up to 80% conversion at high substrate loading (up to 200 mM) without exogenous cofactor addition. The adapted cells displayed longer survival and higher population density during the reactions. The present biotechnological <i>E. coli</i> system contributes to the development of more robust biocatalysis for amine production.","PeriodicalId":25,"journal":{"name":"ACS Sustainable Chemistry & Engineering","volume":null,"pages":null},"PeriodicalIF":7.1000,"publicationDate":"2024-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Amine-Tolerant E. coli Strains Generated via Adaptive Evolution for Sustainable Synthesis of Chiral Amines\",\"authors\":\"Josemarco Mendoza-Avila, Volker Döring, Madeleine Bouzon, Ivan Dubois, Tanja Knaus, Louis M. M. Mouterde, Anne Zaparucha, Francesco G. Mutti, Carine Vergne-Vaxelaire\",\"doi\":\"10.1021/acssuschemeng.4c04356\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The biocatalytic synthesis of chiral amines from carbonyl compounds and ammonia is a major advance in sustainable synthetic chemistry. Using whole cells for bioamination reactions is advantageous given their low preparation cost and direct applicability; however, amine toxicity limits the reaction when living cells are used. Herein, we adapted <i>Escherichia coli</i> BL21(DE3) cells to grow in the presence of 100 mM hexan-2-amine <i>via</i> directed evolution in continuous culture, obtaining six times more tolerant strains than the wild-type. The adapted strains also displayed superior tolerance for structurally different amines. Coexpression of genes encoding for amine dehydrogenase (AmDH) and formate dehydrogenase activities in the adapted strains enabled the stereoselective bioamination (ee > 99%) of different prochiral ketones with up to 80% conversion at high substrate loading (up to 200 mM) without exogenous cofactor addition. The adapted cells displayed longer survival and higher population density during the reactions. The present biotechnological <i>E. coli</i> system contributes to the development of more robust biocatalysis for amine production.\",\"PeriodicalId\":25,\"journal\":{\"name\":\"ACS Sustainable Chemistry & Engineering\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":7.1000,\"publicationDate\":\"2024-09-16\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"ACS Sustainable Chemistry & Engineering\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://doi.org/10.1021/acssuschemeng.4c04356\",\"RegionNum\":1,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Sustainable Chemistry & Engineering","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1021/acssuschemeng.4c04356","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
Amine-Tolerant E. coli Strains Generated via Adaptive Evolution for Sustainable Synthesis of Chiral Amines
The biocatalytic synthesis of chiral amines from carbonyl compounds and ammonia is a major advance in sustainable synthetic chemistry. Using whole cells for bioamination reactions is advantageous given their low preparation cost and direct applicability; however, amine toxicity limits the reaction when living cells are used. Herein, we adapted Escherichia coli BL21(DE3) cells to grow in the presence of 100 mM hexan-2-amine via directed evolution in continuous culture, obtaining six times more tolerant strains than the wild-type. The adapted strains also displayed superior tolerance for structurally different amines. Coexpression of genes encoding for amine dehydrogenase (AmDH) and formate dehydrogenase activities in the adapted strains enabled the stereoselective bioamination (ee > 99%) of different prochiral ketones with up to 80% conversion at high substrate loading (up to 200 mM) without exogenous cofactor addition. The adapted cells displayed longer survival and higher population density during the reactions. The present biotechnological E. coli system contributes to the development of more robust biocatalysis for amine production.
期刊介绍:
ACS Sustainable Chemistry & Engineering is a prestigious weekly peer-reviewed scientific journal published by the American Chemical Society. Dedicated to advancing the principles of green chemistry and green engineering, it covers a wide array of research topics including green chemistry, green engineering, biomass, alternative energy, and life cycle assessment.
The journal welcomes submissions in various formats, including Letters, Articles, Features, and Perspectives (Reviews), that address the challenges of sustainability in the chemical enterprise and contribute to the advancement of sustainable practices. Join us in shaping the future of sustainable chemistry and engineering.