Reconstruction of tyrosol synthetic pathways in Escherichia coli  

作　　者：Cui Yang Xianzhong Chen Junzhuang Chang Lihua Zhang Wei Xu Wei Shen You Fan 

机构地区：[1]Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, Jiangnan University [2]School of Biotechnology, Jiangnan University

出　　处：《Chinese Journal of Chemical Engineering》2018年第12期2615-2621,共7页中国化学工程学报（英文版）

基　　金：Supported by the Fundamental Research Funds for the Central Universities(JUSRP51611A,JUSRP51504);the Natural Science Foundation of Jiangsu Province(BK20171138);the National High Technology Research and Development Program of China(863 program,2013AA102101-5);the 111 Project(No.1112-06)

摘　　要：Tyrosol is a pharmacologically active phenolic compound widely used in the medicine and chemical industries.Traditional methods of plant extraction are complicated and chemical synthesis of tyrosol is not commercially viable. In this study, a recombinant Escherichia coli strain was constructed by overexpressing the phenylpyruvate decarboxylase ARO10 from Saccharomyces cerevisiae, which could produce tyrosol from glucose. Furthermore,genes encoding key enzymes from the competing phenylalanine and tyrosine synthesis pathways and the repression protein TyrR were eliminated, and the resulting engineered strain generated 3.57 mmol·L^(-1) tyrosol from glucose. More significantly, codon optimization of ARO10 increased expression and tyrosol titer. Using the novel engineered strain expressing codon-optimized AR10 in shake-flask culture, 8.72 mmol·L^(-1) tyrosol was obtained after 48 h. Optimization of the induction conditions improved tyrosol production to 9.53 mmol·L^(-1)(1316.3 mg·L^(-1)). A higher titer of tyrosol was achieved by reconstruction of tyrosol synthetic pathway in E. coli.Tyrosol is a pharmacologically active phenolic compound widely used in the medicine and chemical industries.Traditional methods of plant extraction are complicated and chemical synthesis of tyrosol is not commercially viable. In this study, a recombinant Escherichia coli strain was constructed by overexpressing the phenylpyruvate decarboxylase ARO10 from Saccharomyces cerevisiae, which could produce tyrosol from glucose. Furthermore,genes encoding key enzymes from the competing phenylalanine and tyrosine synthesis pathways and the repression protein TyrR were eliminated, and the resulting engineered strain generated 3.57 mmol·L^(-1) tyrosol from glucose. More significantly, codon optimization of ARO10 increased expression and tyrosol titer. Using the novel engineered strain expressing codon-optimized AR10 in shake-flask culture, 8.72 mmol·L^(-1) tyrosol was obtained after 48 h. Optimization of the induction conditions improved tyrosol production to 9.53 mmol·L^(-1)(1316.3 mg·L^(-1)). A higher titer of tyrosol was achieved by reconstruction of tyrosol synthetic pathway in E. coli.

关 键 词：TYROSOL ESCHERICHIA COLI Phenylpyruvate DECARBOXYLASE Gene KNOCKOUT CODON optimization 

分 类 号：TQ[化学工程]