SCI和EI收录∣中国化工学会会刊

Chinese Journal of Chemical Engineering ›› 2018, Vol. 26 ›› Issue (12): 2615-2621.DOI: 10.1016/j.cjche.2018.04.020

• Biotechnology and Bioengineering • 上一篇    下一篇

Reconstruction of tyrosol synthetic pathways in Escherichia coli

Cui Yang1,2, Xianzhong Chen1,2, Junzhuang Chang1,2, Lihua Zhang1,2, Wei Xu1,2, Wei Shen1, You Fan1,2   

  1. 1 Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;
    2 School of Biotechnology, Jiangnan University, Wuxi 214122, China
  • 收稿日期:2017-11-23 修回日期:2018-03-17 出版日期:2018-12-28 发布日期:2019-01-09
  • 通讯作者: Xianzhong Chen
  • 基金资助:

    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) and the 111 Project (No. 1112-06).

Reconstruction of tyrosol synthetic pathways in Escherichia coli

Cui Yang1,2, Xianzhong Chen1,2, Junzhuang Chang1,2, Lihua Zhang1,2, Wei Xu1,2, Wei Shen1, You Fan1,2   

  1. 1 Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;
    2 School of Biotechnology, Jiangnan University, Wuxi 214122, China
  • Received:2017-11-23 Revised:2018-03-17 Online:2018-12-28 Published:2019-01-09
  • Contact: Xianzhong Chen
  • Supported by:

    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) and 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.

关键词: Tyrosol, Escherichia coli, Phenylpyruvate decarboxylase, Gene knockout, Codon optimization

Abstract: 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.

Key words: Tyrosol, Escherichia coli, Phenylpyruvate decarboxylase, Gene knockout, Codon optimization