Claudia Harting, Attila Teleki, Marius Braakmann, Frank Jankowitsch, Ralf Takors
{"title":"在转录失调的大肠杆菌蛋氨酸生产者中平衡复合生物合成的细胞内系统分析。","authors":"Claudia Harting, Attila Teleki, Marius Braakmann, Frank Jankowitsch, Ralf Takors","doi":"10.1111/1751-7915.14433","DOIUrl":null,"url":null,"abstract":"<p><span>l</span>-Methionine (<span>l</span>-Met) has gained remarkable interest due to its multifaceted and versatile applications in the fields of nutrition, pharmaceuticals and clinical practice. In this study, the fluxes of the challenging <span>l</span>-Met biosynthesis in the producer strain <i>Escherichia coli</i> (<i>E. coli</i>) DM2853 were fine-tuned to enable improved <span>l</span>-Met production. The potential bottlenecks identified in sulfur assimilation and <span>l</span>-Met synthesis downstream of <i>O</i>-succinyl-<span>l</span>-homoserine (OSHS) were addressed by overexpressing glutaredoxin 1 (<i>grxA</i>), thiosulfate sulfurtransferase (<i>pspE</i>) and <i>O</i>-succinylhomoserine lyase (<i>metB</i>). Although deemed as a straightforward target for improving glucose-to-Met conversion, the yields remained at approximately 12%–13% (g/g). Instead, intracellular <span>l</span>-Met pools increased by up to four-fold with accelerated kinetics. Overexpression of the Met exporter <i>ygaZH</i> may serve as a proper valve for releasing the rising internal Met pressure. Interestingly, the export kinetics revealed maximum saturated export rates already at low growth rates. This scenario is particularly advantageous for large-scale fermentation when product formation is ideally uncoupled from biomass formation to achieve maximum performance within the technical limits of large-scale bioreactors.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":null,"pages":null},"PeriodicalIF":5.7000,"publicationDate":"2024-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.14433","citationCount":"0","resultStr":"{\"title\":\"Systemic intracellular analysis for balancing complex biosynthesis in a transcriptionally deregulated Escherichia coli l-Methionine producer\",\"authors\":\"Claudia Harting, Attila Teleki, Marius Braakmann, Frank Jankowitsch, Ralf Takors\",\"doi\":\"10.1111/1751-7915.14433\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><span>l</span>-Methionine (<span>l</span>-Met) has gained remarkable interest due to its multifaceted and versatile applications in the fields of nutrition, pharmaceuticals and clinical practice. In this study, the fluxes of the challenging <span>l</span>-Met biosynthesis in the producer strain <i>Escherichia coli</i> (<i>E. coli</i>) DM2853 were fine-tuned to enable improved <span>l</span>-Met production. The potential bottlenecks identified in sulfur assimilation and <span>l</span>-Met synthesis downstream of <i>O</i>-succinyl-<span>l</span>-homoserine (OSHS) were addressed by overexpressing glutaredoxin 1 (<i>grxA</i>), thiosulfate sulfurtransferase (<i>pspE</i>) and <i>O</i>-succinylhomoserine lyase (<i>metB</i>). Although deemed as a straightforward target for improving glucose-to-Met conversion, the yields remained at approximately 12%–13% (g/g). Instead, intracellular <span>l</span>-Met pools increased by up to four-fold with accelerated kinetics. Overexpression of the Met exporter <i>ygaZH</i> may serve as a proper valve for releasing the rising internal Met pressure. Interestingly, the export kinetics revealed maximum saturated export rates already at low growth rates. This scenario is particularly advantageous for large-scale fermentation when product formation is ideally uncoupled from biomass formation to achieve maximum performance within the technical limits of large-scale bioreactors.</p>\",\"PeriodicalId\":209,\"journal\":{\"name\":\"Microbial Biotechnology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.7000,\"publicationDate\":\"2024-03-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1111/1751-7915.14433\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbial Biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/1751-7915.14433\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/1751-7915.14433","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Systemic intracellular analysis for balancing complex biosynthesis in a transcriptionally deregulated Escherichia coli l-Methionine producer
l-Methionine (l-Met) has gained remarkable interest due to its multifaceted and versatile applications in the fields of nutrition, pharmaceuticals and clinical practice. In this study, the fluxes of the challenging l-Met biosynthesis in the producer strain Escherichia coli (E. coli) DM2853 were fine-tuned to enable improved l-Met production. The potential bottlenecks identified in sulfur assimilation and l-Met synthesis downstream of O-succinyl-l-homoserine (OSHS) were addressed by overexpressing glutaredoxin 1 (grxA), thiosulfate sulfurtransferase (pspE) and O-succinylhomoserine lyase (metB). Although deemed as a straightforward target for improving glucose-to-Met conversion, the yields remained at approximately 12%–13% (g/g). Instead, intracellular l-Met pools increased by up to four-fold with accelerated kinetics. Overexpression of the Met exporter ygaZH may serve as a proper valve for releasing the rising internal Met pressure. Interestingly, the export kinetics revealed maximum saturated export rates already at low growth rates. This scenario is particularly advantageous for large-scale fermentation when product formation is ideally uncoupled from biomass formation to achieve maximum performance within the technical limits of large-scale bioreactors.
期刊介绍:
Microbial Biotechnology publishes papers of original research reporting significant advances in any aspect of microbial applications, including, but not limited to biotechnologies related to: Green chemistry; Primary metabolites; Food, beverages and supplements; Secondary metabolites and natural products; Pharmaceuticals; Diagnostics; Agriculture; Bioenergy; Biomining, including oil recovery and processing; Bioremediation; Biopolymers, biomaterials; Bionanotechnology; Biosurfactants and bioemulsifiers; Compatible solutes and bioprotectants; Biosensors, monitoring systems, quantitative microbial risk assessment; Technology development; Protein engineering; Functional genomics; Metabolic engineering; Metabolic design; Systems analysis, modelling; Process engineering; Biologically-based analytical methods; Microbially-based strategies in public health; Microbially-based strategies to influence global processes