{"title":"葡萄糖高产l -苯丙氨酸重组大肠杆菌的构建","authors":"Pengfei Gu, Shuo Zhao, Chengwei Li, Shuixing Jiang, Hao Zhou, Qiang Li","doi":"10.3390/microbiolres14030079","DOIUrl":null,"url":null,"abstract":"L-phenylalanine is an important aromatic amino acid that is widely used in the area of feed, food additives, and pharmaceuticals. Among the different strategies of L-phenylalanine synthesis, direct microbial fermentation from raw substrates has attracted more and more attention due to its environment friendly process and low-cost raw materials. In this study, a rational designed recombinant Escherichia coli was constructed for L-phenylalanine production. Based on wild type E. coli MG1655, multilevel engineering strategies were carried out, such as directing more carbon flux into the L-phenylalanine synthetic pathway, increasing intracellular level of precursors, blocking by-product synthesis pathways and facilitating the secretion of L-phenylalanine. During 5 L fed batch fermentation, recombinant E. coli MPH-3 could produce 19.24 g/L of L-phenylalanine with a yield of 0.279 g/g glucose. To the best of our knowledge, this is one of the highest yields of L-phenylalanine producing E. coli using glucose as the sole carbon source in fed-batch fermentation.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"2 1","pages":""},"PeriodicalIF":2.1000,"publicationDate":"2023-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Construction of Recombinant Escherichia coli with a High L-Phenylalanine Production Yield from Glucose\",\"authors\":\"Pengfei Gu, Shuo Zhao, Chengwei Li, Shuixing Jiang, Hao Zhou, Qiang Li\",\"doi\":\"10.3390/microbiolres14030079\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"L-phenylalanine is an important aromatic amino acid that is widely used in the area of feed, food additives, and pharmaceuticals. Among the different strategies of L-phenylalanine synthesis, direct microbial fermentation from raw substrates has attracted more and more attention due to its environment friendly process and low-cost raw materials. In this study, a rational designed recombinant Escherichia coli was constructed for L-phenylalanine production. Based on wild type E. coli MG1655, multilevel engineering strategies were carried out, such as directing more carbon flux into the L-phenylalanine synthetic pathway, increasing intracellular level of precursors, blocking by-product synthesis pathways and facilitating the secretion of L-phenylalanine. During 5 L fed batch fermentation, recombinant E. coli MPH-3 could produce 19.24 g/L of L-phenylalanine with a yield of 0.279 g/g glucose. To the best of our knowledge, this is one of the highest yields of L-phenylalanine producing E. coli using glucose as the sole carbon source in fed-batch fermentation.\",\"PeriodicalId\":43788,\"journal\":{\"name\":\"Microbiology Research\",\"volume\":\"2 1\",\"pages\":\"\"},\"PeriodicalIF\":2.1000,\"publicationDate\":\"2023-08-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbiology Research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.3390/microbiolres14030079\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbiology Research","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3390/microbiolres14030079","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
l -苯丙氨酸是一种重要的芳香氨基酸,广泛应用于饲料、食品添加剂和药品等领域。在各种合成l -苯丙氨酸的方法中,以原料为原料的微生物直接发酵因其工艺环境友好、原料成本低而受到越来越多的关注。本研究构建了一种合理设计的重组大肠杆菌,用于生产l -苯丙氨酸。以野生型大肠杆菌MG1655为基础,进行了引导更多碳通量进入l -苯丙氨酸合成途径、增加细胞内前体水平、阻断副产物合成途径、促进l -苯丙氨酸分泌等多层次工程策略。在5 L补料间歇发酵过程中,重组大肠杆菌MPH-3的L-苯丙氨酸产率为19.24 g/L,葡萄糖产率为0.279 g/g。据我们所知,这是在饲料分批发酵中使用葡萄糖作为唯一碳源的l -苯丙氨酸生产大肠杆菌的最高产量之一。
Construction of Recombinant Escherichia coli with a High L-Phenylalanine Production Yield from Glucose
L-phenylalanine is an important aromatic amino acid that is widely used in the area of feed, food additives, and pharmaceuticals. Among the different strategies of L-phenylalanine synthesis, direct microbial fermentation from raw substrates has attracted more and more attention due to its environment friendly process and low-cost raw materials. In this study, a rational designed recombinant Escherichia coli was constructed for L-phenylalanine production. Based on wild type E. coli MG1655, multilevel engineering strategies were carried out, such as directing more carbon flux into the L-phenylalanine synthetic pathway, increasing intracellular level of precursors, blocking by-product synthesis pathways and facilitating the secretion of L-phenylalanine. During 5 L fed batch fermentation, recombinant E. coli MPH-3 could produce 19.24 g/L of L-phenylalanine with a yield of 0.279 g/g glucose. To the best of our knowledge, this is one of the highest yields of L-phenylalanine producing E. coli using glucose as the sole carbon source in fed-batch fermentation.
期刊介绍:
Microbiology Research is an international, online-only, open access peer-reviewed journal which publishes original research, review articles, editorials, perspectives, case reports and brief reports to benefit researchers, microbiologists, physicians, veterinarians. Microbiology Research publishes ‘Clinic’ and ‘Research’ papers divided into two different skill and proficiency levels: ‘Junior’ and ‘Professional’. The aim of this four quadrant grid is to encourage younger researchers, physicians and veterinarians to submit their results even if their studies encompass just a limited set of observations or rely on basic statistical approach, yet upholding the customary sound approach of every scientific article.