Qin Ke, Chang Liu, Yibin Zhuang, Yaju Xue, Zhanzhao Cui, Cuiying Zhang, Hua Yin, Tao Liu
{"title":"Metabolic engineering of Escherichia coli for high-level production of benzyl acetate from glucose.","authors":"Qin Ke, Chang Liu, Yibin Zhuang, Yaju Xue, Zhanzhao Cui, Cuiying Zhang, Hua Yin, Tao Liu","doi":"10.1186/s12934-024-02513-y","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Benzyl acetate is an aromatic ester with a jasmine scent. It was discovered in plants and has broad applications in food, cosmetic, and pharmaceutical industries. Its current production predominantly relies on chemical synthesis. In this study, Escherichia coli was engineered to produce benzyl acetate.</p><p><strong>Results: </strong>Two biosynthetic routes based on the CoA-dependent β-oxidation pathway were constructed in E. coli for benzyl acetate production. In route I, benzoic acid pathway was extended to produce benzyl alcohol by combining carboxylic acid reductase and endogenous dehydrogenases and/or aldo-keto reductases in E. coli. Benzyl alcohol was then condensed with acetyl-CoA by the alcohol acetyltransferase ATF1 from yeast to form benzyl acetate. In route II, a plant CoA-dependent β-oxidation pathway via benzoyl-CoA was assessed for benzyl alcohol and benzyl acetate production in E. coli. The overexpression of the phosphotransacetylase from Clostridium kluyveri (CkPta) further improved benzyl acetate production in E. coli. Two-phase extractive fermentation in situ was adopted and optimized for benzyl acetate production in a shake flask. The most optimal strain produced 3.0 ± 0.2 g/L benzyl acetate in 48 h by shake-flask fermentation.</p><p><strong>Conclusions: </strong>We were able to establish the whole pathway for benzyl acetate based on the CoA-dependent β-oxidation in single strain for the first time. The highest titer for benzyl acetate produced from glucose by E. coli is reported. Moreover, cinnamyl acetate production as an unwanted by-product was very low. Results provided novel information regarding the engineering benzyl acetate production in microorganisms.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":null,"pages":null},"PeriodicalIF":4.3000,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11370050/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial Cell Factories","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1186/s12934-024-02513-y","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Benzyl acetate is an aromatic ester with a jasmine scent. It was discovered in plants and has broad applications in food, cosmetic, and pharmaceutical industries. Its current production predominantly relies on chemical synthesis. In this study, Escherichia coli was engineered to produce benzyl acetate.
Results: Two biosynthetic routes based on the CoA-dependent β-oxidation pathway were constructed in E. coli for benzyl acetate production. In route I, benzoic acid pathway was extended to produce benzyl alcohol by combining carboxylic acid reductase and endogenous dehydrogenases and/or aldo-keto reductases in E. coli. Benzyl alcohol was then condensed with acetyl-CoA by the alcohol acetyltransferase ATF1 from yeast to form benzyl acetate. In route II, a plant CoA-dependent β-oxidation pathway via benzoyl-CoA was assessed for benzyl alcohol and benzyl acetate production in E. coli. The overexpression of the phosphotransacetylase from Clostridium kluyveri (CkPta) further improved benzyl acetate production in E. coli. Two-phase extractive fermentation in situ was adopted and optimized for benzyl acetate production in a shake flask. The most optimal strain produced 3.0 ± 0.2 g/L benzyl acetate in 48 h by shake-flask fermentation.
Conclusions: We were able to establish the whole pathway for benzyl acetate based on the CoA-dependent β-oxidation in single strain for the first time. The highest titer for benzyl acetate produced from glucose by E. coli is reported. Moreover, cinnamyl acetate production as an unwanted by-product was very low. Results provided novel information regarding the engineering benzyl acetate production in microorganisms.
背景:乙酸苄酯是一种具有茉莉香味的芳香酯。它是在植物中发现的,在食品、化妆品和制药行业有着广泛的应用。目前乙酸苄酯的生产主要依靠化学合成。在这项研究中,我们设计了大肠杆菌来生产乙酸苄酯:结果:在大肠杆菌中构建了两条基于 CoA 依赖性β-氧化途径的生物合成路线,用于生产乙酸苄酯。在途径 I 中,通过结合大肠杆菌中的羧酸还原酶和内源脱氢酶和/或醛酮还原酶,苯甲酸途径被扩展为生产苄醇。然后,酵母中的乙醇乙酰转移酶 ATF1 将苄醇与乙酰-CoA 缩合,形成乙酸苄酯。在路线 II 中,通过苯甲酰-CoA 的植物 CoA 依赖性 β 氧化途径,对大肠杆菌生产苄醇和乙酸苄酯进行了评估。过量表达来自克鲁伊韦氏梭菌(CkPta)的磷酸反式乙酰化酶进一步提高了大肠杆菌的乙酸苄酯产量。在摇瓶中采用两相原位萃取发酵法生产乙酸苄酯,并进行了优化。通过摇瓶发酵,最优菌株在 48 小时内生产出 3.0 ± 0.2 克/升乙酸苄酯:结论:我们首次在单一菌株中建立了基于 CoA 依赖性 β 氧化的乙酸苄酯生产全过程。报告了大肠杆菌从葡萄糖中产生乙酸苄酯的最高滴度。此外,作为无用副产物的乙酸肉桂酯的产量也非常低。研究结果提供了有关微生物乙酸苄酯生产工程的新信息。
期刊介绍:
Microbial Cell Factories is an open access peer-reviewed journal that covers any topic related to the development, use and investigation of microbial cells as producers of recombinant proteins and natural products, or as catalyzers of biological transformations of industrial interest. Microbial Cell Factories is the world leading, primary research journal fully focusing on Applied Microbiology.
The journal is divided into the following editorial sections:
-Metabolic engineering
-Synthetic biology
-Whole-cell biocatalysis
-Microbial regulations
-Recombinant protein production/bioprocessing
-Production of natural compounds
-Systems biology of cell factories
-Microbial production processes
-Cell-free systems