Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli

doi:10.1016/j.cjche.2021.05.026

Chinese Journal of Chemical Engineering ›› 2022, Vol. 47 ›› Issue (7): 18-30.DOI: 10.1016/j.cjche.2021.05.026

Previous Articles Next Articles

Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli

Xiaobo Ruan^1,2, Sheng Zhang³, Wei Song^1,2, Jia Liu², Xiulai Chen², Liming Liu², Jing Wu¹

1. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China;
2. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;
3. Tianrui Chemical Co., Ltd, Department of Chemistry, Quzhou 324400, China

Received:2021-02-06 Revised:2021-05-02 Online:2022-08-19 Published:2022-07-28
Contact: Jing Wu,E-mail:wujing@jiangnan.edu.cn
Supported by:
This work was financially supported by the Fundamental Research Funds for the Central Universities (JUSRP21915), National Natural Science Foundation of China (22008089, 21878126), Provincial Natural Science Foundation of Jiangsu Province (BK20200622), the key technologies Research & Development Program of Jiangsu Province (BE2018623), and the National First-Class Discipline Program of Light Industry Technology and Engineering (LITE2018-20).

Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli

Xiaobo Ruan^1,2, Sheng Zhang³, Wei Song^1,2, Jia Liu², Xiulai Chen², Liming Liu², Jing Wu¹

1. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China;
2. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;
3. Tianrui Chemical Co., Ltd, Department of Chemistry, Quzhou 324400, China

通讯作者: Jing Wu,E-mail:wujing@jiangnan.edu.cn
基金资助:
This work was financially supported by the Fundamental Research Funds for the Central Universities (JUSRP21915), National Natural Science Foundation of China (22008089, 21878126), Provincial Natural Science Foundation of Jiangsu Province (BK20200622), the key technologies Research & Development Program of Jiangsu Province (BE2018623), and the National First-Class Discipline Program of Light Industry Technology and Engineering (LITE2018-20).

Abstract

Abstract: For the efficient conversion of L-tyrosine (L-Tyr) to tyrosol, which is an aromatic compound widely used in the pharmaceutical and chemical industries, a novel four-enzyme cascade pathway based on the Ehrlich pathway of Saccharomyces cerevisiae was designed and reconstructed in Escherichia coli. Then, the expression levels of the relevant enzymes were coordinated using a modular approach and gene duplication after the identification of the pyruvate decarboxylase from Candida tropicalis (CtPDC) as the rate-limiting enzymatic step. In situ product removal (ISPR) strategy with XAD4 resins was explored to avoid product inhibition and further improve tyrosol yield. As a result, the titer and conversion rate of tyrosol obtained were 35.7 g·L^-1 and 93.6%, respectively, in a 3-L bioreactor. Results presented here provide a potential enzymatic process for industrial production of tyrosol from cheap amino acids.

Key words: Tyrosol, L-tyrosine, Ehrlich pathway, Enzyme cascade, In situ product removal

摘要： For the efficient conversion of L-tyrosine (L-Tyr) to tyrosol, which is an aromatic compound widely used in the pharmaceutical and chemical industries, a novel four-enzyme cascade pathway based on the Ehrlich pathway of Saccharomyces cerevisiae was designed and reconstructed in Escherichia coli. Then, the expression levels of the relevant enzymes were coordinated using a modular approach and gene duplication after the identification of the pyruvate decarboxylase from Candida tropicalis (CtPDC) as the rate-limiting enzymatic step. In situ product removal (ISPR) strategy with XAD4 resins was explored to avoid product inhibition and further improve tyrosol yield. As a result, the titer and conversion rate of tyrosol obtained were 35.7 g·L^-1 and 93.6%, respectively, in a 3-L bioreactor. Results presented here provide a potential enzymatic process for industrial production of tyrosol from cheap amino acids.

关键词: Tyrosol, L-tyrosine, Ehrlich pathway, Enzyme cascade, In situ product removal

Xiaobo Ruan, Sheng Zhang, Wei Song, Jia Liu, Xiulai Chen, Liming Liu, Jing Wu. Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli[J]. Chinese Journal of Chemical Engineering, 2022, 47(7): 18-30.

References

[1] D. Chung, S.Y. Kim, J.H. Ahn, Production of three phenylethanoids, tyrosol, hydroxytyrosol, and salidroside, using plant genes expressing in Escherichia coli, Sci. Rep. 7(2017)2578
[2] P. Dewapriya, Y.X. Li, S.W.A. Himaya, S.K. Kim, Isolation and characterization of marine-derived Mucor sp. for the fermentative production of tyrosol, Process Biochem. 49(2014)1402-1408
[3] H. Soejima, K. Tsuge, T. Yoshimura, K. Sawada, H. Kitagaki, Breeding of a high tyrosol-producing sake yeast by isolation of an ethanol-resistant mutant from a trp3 mutant, J. Inst. Brew. 118(2012)264-268
[4] L.R. Silva, P.B. Andrade, P. Valentão, R.M. Seabra, M.E. Trujillo, E. Velázquez, Analysis of non-coloured phenolics in red wine:Effect of Dekkera bruxellensis yeast, Food Chem. 89(2005)185-189
[5] W. Guo, Q. Huang, H. Liu, S. Hou, S. Niu, Y. Jiang, X. Bao, Y. Shen, X. Fang, Rational engineering of chorismate-related pathways in Saccharomyces cerevisiae for improving tyrosol production, Front. Bioeng. Biotechnol. 7(2019)152
[6] H. Yang, Y. Xue, C. Yang, W. Shen, Y. Fan, X. Chen, Modular engineering of tyrosol production in Escherichia coli, J. Agric. Food Chem. 67(2019)3900-3908
[7] H.M. Woodburn, C.F. Stuntz, The synthesis of β-(3-amino-4-hydroxyphenyl)-ethanol; 3-aminotyrosol, J. Am. Chem. Soc. 72(1950)1361-1364
[8] S.E. Yoo, Y.D. Gong, M.Y. Choi, J. Seo, A new vinyl ether type linker for solid-phase synthesis, Tetrahedron Lett. 41(2000)6415-6418
[9] S. Yamada,T. Fujii, K. Takagi, Y. Gomi, Preparation of tyrosol and 4-methoxyphenethyl alcohol, Chem. Pharm. Bull. 11(1963)258-260
[10] S. Sentheshanmuganathan, S.R. Elsden, The mechanism of the formation of tyrosol by Saccharomyces cerevisiae, J. Biochem. 69(1958)210-218
[11] J. Jiang, H. Yin, S. Wang, Y. Zhuang, S. Liu, T. Liu, Y. Ma, Metabolic engineering of Saccharomyces cerevisiae for high-level production of salidroside from glucose, J. Agric. Food Chem. 66(2018)4431-4438
[12] W. Guo, Q. Huang, Y. Feng, T. Tan, S. Niu, S. Hou, Z. Chen, Z.Q. Du, Y. Shen, X. Fang, Rewiring central carbon metabolism for tyrosol and salidroside production in Saccharomyces cerevisiae, Biotechnol. Bioeng. 117(2020)2410-2419
[13] H. Liu, Y. Tian, Y. Zhou, Y. Kan, T. Wu, W. Xiao, Y. Luo, Multi-modular engineering of Saccharomyces cerevisiae for high-titre production of tyrosol and salidroside, Microb. Biotechnol.(2020)1-12
[14] Y. Satoh, K. Tajima, M. Munekata, J.D. Keasling, T.S. Lee, Engineering of a tyrosol-producing pathway, utilizing simple sugar and the central metabolic tyrosine, in Escherichia coli, J. Agric. Food Chem. 60(2012)979-984
[15] L.A. Hazelwood, J.M. Daran, A.J.A. Van Maris, J.T. Pronk, J.R. Dickinson, The Ehrlich pathway for fusel alcohol production:A century of research on Saccharomyces cerevisiae metabolism, Appl. Environ. Microbiol. 74(2008)2259-2266
[16] Y. Kaminaga, J. Schnepp, G. Peel, C.M. Kish, G. Ben-Nissan, Plant phenylacetaldehyde synthase is a bifunctional homotetrameric enzyme that catalyzes phenylalanine decarboxylation and oxidation, J. Biol. Chem. 281(2006)23357-23366
[17] Y. Xue, X. Chen, C. Yang, J. Chang, W. Shen, Y. Fan, Engineering Eschericha coli for enhanced tyrosol production, J. Agric. Food Chem. 65(2017)4708-4714
[18] C. Yang, X. Chen, J. Chang, L. Zhang, W. Xu, W. Shen, Y. Fan, Reconstruction of tyrosol synthetic pathways in Escherichia coli, Chinese J. Chem. Eng. 26(2018)2615-2621
[19] W. Xu, C. Yang, Y. Xia, L. Zhang, C. Liu, H. Yang, W. Shen, X. Chen, High-level production of tyrosol with noninduced recombinant Escherichia coli by metabolic engineering, J. Agric. Food Chem. 68(2020)4616-4623
[20] Z. Mao, L. Liu, Y. Zhang, J. Yuan, Efficient synthesis of phenylacetate and 2-phenylethanol by modular cascade biocatalysis, ChemBioChem 21(2020)2676-2679
[21] F. Zhu, X. Zhong, M. Hu, L. Lu, Z. Deng, T. Liu, In vitro reconstitution of mevalonate pathway and targeted engineering of farnesene overproduction in Escherichia coli, Biotechnol. Bioeng. 111(2014)1396-1405
[22] C. Li, P. Jia, Y. Bai, T.P. Fan, X. Zheng, Y. Cai, Efficient synthesis of hydroxytyrosol from L-3,4-dihydroxyphenylalanine using engineered Escherichia coli whole cells, J. Agric. Food Chem. 67(2019)6867-6873
[23] G. Molla, R. Melis, L. Pollegioni, Breaking the mirror:L-amino acid deaminase, a novel stereoselective biocatalyst, Biotechnol. Adv. 35(2017)657-668
[24] C. Rodriguez, I. Lavandera, V. Gotor, Recent advances in cofactor regeneration systems applied to biocatalyzed oxidative processes, Curr. Org. Chem. 16(2012)2525-2541
[25] J. Wu, G. Du, J. Zhou, J. Chen, Metabolic engineering of Escherichia coli for (2S)-pinocembrin production from glucose by a modular metabolic strategy, Metab. Eng. 16(2013)48-55
[26] J. Wang, W. Song, J. Wu, J. Liu, X. Chen, L. Liu, Efficient production of phenylpropionic acids by an amino-group-transformation biocatalytic cascade, Biotechnol. Bioeng. 117(2020)614-625
[27] B.R. Lukito, S. Wu, H.J.J. Saw, Z. Li, One-pot production of natural 2-phenylethanol from L-phenylalanine via cascade biotransformations, ChemCatChem 11(2019)831-840
[28] J. Yao, Y. He, N. Su, S.R. Bharath, Y. Tao, J.M. Jin, W. Chen, H. Song, S.Y. Tang, Developing a highly efficient hydroxytyrosol whole-cell catalyst by de-bottlenecking rate-limiting steps, Nat. Commun. 11(2020)1-12
[29] Y. Qian, J. Liu, W. Song, X. Chen, Q. Luo, L. Liu, Production of β-alanine from fumaric acid using a dual-enzyme cascade, ChemCatChem 10(2018)4998-5005
[30] J.J. Lucchini, J. Corre, A. Cremieux, Antibacterial activity of phenolic compounds and aromatic alcohols, Res. Microbiol. 141(1990)499-510
[31] S.H. Liu, I.H. Pan, I.M. Chu, Inhibitory effect of p-hydroxybenzyl alcohol on tyrosinase activity and melanogenesis, Biol. Pharm. Bull. 30(2007)1135-1139
[32] A.G. Santos, T.L. de Albuquerque, B.D. Ribeiro, M.A.Z. Coelho, In situ product recovery techniques aiming to obtain biotechnological products:A glance to current knowledge, Biotechnol. Appl. Biochem.(2020)1-14
[33] M.E. Hong, K.S. Lee, B.J. Yu, Y.J. Sung, S.M. Park, H.M. Koo, Identification of gene targets eliciting improved alcohol tolerance in Saccharomyces cerevisiae through inverse metabolic engineering, J. Biotechnol. 149(2010)52-59
[34] D. Hua, S. Lin, Y. Li, H. Chen, Z. Zhang, Y. Du, X. Zhang, P. Xu, Enhanced 2-phenylethanol production from L-phenylalanine via in situ product adsorption, Biocatal. Biotransfor. 28(2010)259-266
[35] D.R. Nielsen, K.J. Prather, In situ product recovery of n-butanol using polymeric resins, Biotechnol. Bioeng. 102(2009)811-821

Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli

Efficient synthesis of tyrosol from L-tyrosine via heterologous Ehrlich pathway in Escherichia coli

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 3

Recommended Articles

Metrics

Comments

[1]	Xiaohong Zhou, Wenfeng Zhou, Wei Zhuang, Chenjie Zhu, Hanjie Ying, Hongman Zhang. Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose [J]. Chinese Journal of Chemical Engineering, 2023, 58(6): 40-52.
[2]	Huanru Ding, Weirui Zhao, Changjiang Lü, Jun Huang, Sheng Hu, Shanjing Yao, Lehe Mei, Jinbo Wang, Jiaqi Mei. Biosynthesis of 4-hydroxyphenylpyruvic acid from L-tyrosine using recombinant Escherichia coli cells expressing membrane bound L-amino acid deaminase [J]. Chin.J.Chem.Eng., 2018, 26(2): 380-385.
[3]	Cui Yang, Xianzhong Chen, Junzhuang Chang, Lihua Zhang, Wei Xu, Wei Shen, You Fan. Reconstruction of tyrosol synthetic pathways in Escherichia coli [J]. Chin.J.Chem.Eng., 2018, 26(12): 2615-2621.