Cell-free protein synthesis enables one-pot cascade biotransformation in an aqueous-organic biphasic system

Wan Qiu Liu; Changzhu Wu; Michael C. Jewett; Jian Li

doi:10.1002/bit.27541

Cell-free protein synthesis enables one-pot cascade biotransformation in an aqueous-organic biphasic system

Wan Qiu Liu, Changzhu Wu^*, Michael C. Jewett, Jian Li^*

^*Corresponding author for this work

Chemical and Biological Engineering

Research output: Contribution to journal › Article › peer-review

28 Scopus citations

Abstract

Biocatalytic cascade reactions have become increasingly important and useful for chemical synthesis. However, biocatalysts are often incompatible with organic solvents, which prohibits many cascade reactions involving nonpolar substrates. In this study, we used cell-free protein synthesis (CFPS) to express enzymes in an aqueous-organic biphasic system for the construction of an artificial enzymatic pathway. CFPS-expressed enzymes without purification performed efficiently to convert styrene (below 20 mM) to (S)-1-phenyl-1,2-ethanediol (two steps in one pot) with 100% conversion. In addition, our CFPS system showed great tolerance to different organic solvents, and, importantly, the entire biocatalytic system can be consistently scaled up without a reduction of the substrate conversion rate. We, therefore, anticipate that our cell-free approach will make a possible cost-effective, high-yielding synthesis of valuable chemicals.

Original language	English (US)
Pages (from-to)	4001-4008
Number of pages	8
Journal	Biotechnology and Bioengineering
Volume	117
Issue number	12
DOIs	https://doi.org/10.1002/bit.27541
State	Published - Dec 2020

Funding

The authors would like to acknowledge Ashty Karim for helpful discussions. This study was supported by grants from the National Natural Science Foundation of China (Grant Nos. 31971348 and 31800720), the Natural Science Foundation of Shanghai (Grant No. 19ZR1477200), and the Shanghai Pujiang Program (Grant No. 18PJ1408000). J. L. also thanks the starting grant of ShanghaiTech University. M. C. J. acknowledges support from the Department of Energy Grant DE‐SC0018249, the David and Lucile Packard Foundation, and the Camille Dreyfus Teacher‐Scholar Program. The authors would like to acknowledge Ashty Karim for helpful discussions. This study was supported by grants from the National Natural Science Foundation of China (Grant Nos. 31971348 and 31800720), the Natural Science Foundation of Shanghai (Grant No. 19ZR1477200), and the Shanghai Pujiang Program (Grant No. 18PJ1408000). J. L. also thanks the starting grant of ShanghaiTech University. M. C. J. acknowledges support from the Department of Energy Grant DE-SC0018249, the David and Lucile Packard Foundation, and the Camille Dreyfus Teacher-Scholar Program.

Keywords

biocatalysis
biotransformation
biphasic system
cascade reaction
cell-free protein synthesis

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1002/bit.27541

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title = "Cell-free protein synthesis enables one-pot cascade biotransformation in an aqueous-organic biphasic system",

abstract = "Biocatalytic cascade reactions have become increasingly important and useful for chemical synthesis. However, biocatalysts are often incompatible with organic solvents, which prohibits many cascade reactions involving nonpolar substrates. In this study, we used cell-free protein synthesis (CFPS) to express enzymes in an aqueous-organic biphasic system for the construction of an artificial enzymatic pathway. CFPS-expressed enzymes without purification performed efficiently to convert styrene (below 20 mM) to (S)-1-phenyl-1,2-ethanediol (two steps in one pot) with 100% conversion. In addition, our CFPS system showed great tolerance to different organic solvents, and, importantly, the entire biocatalytic system can be consistently scaled up without a reduction of the substrate conversion rate. We, therefore, anticipate that our cell-free approach will make a possible cost-effective, high-yielding synthesis of valuable chemicals.",

keywords = "biocatalysis, biotransformation, biphasic system, cascade reaction, cell-free protein synthesis",

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Y1 - 2020/12

N2 - Biocatalytic cascade reactions have become increasingly important and useful for chemical synthesis. However, biocatalysts are often incompatible with organic solvents, which prohibits many cascade reactions involving nonpolar substrates. In this study, we used cell-free protein synthesis (CFPS) to express enzymes in an aqueous-organic biphasic system for the construction of an artificial enzymatic pathway. CFPS-expressed enzymes without purification performed efficiently to convert styrene (below 20 mM) to (S)-1-phenyl-1,2-ethanediol (two steps in one pot) with 100% conversion. In addition, our CFPS system showed great tolerance to different organic solvents, and, importantly, the entire biocatalytic system can be consistently scaled up without a reduction of the substrate conversion rate. We, therefore, anticipate that our cell-free approach will make a possible cost-effective, high-yielding synthesis of valuable chemicals.

AB - Biocatalytic cascade reactions have become increasingly important and useful for chemical synthesis. However, biocatalysts are often incompatible with organic solvents, which prohibits many cascade reactions involving nonpolar substrates. In this study, we used cell-free protein synthesis (CFPS) to express enzymes in an aqueous-organic biphasic system for the construction of an artificial enzymatic pathway. CFPS-expressed enzymes without purification performed efficiently to convert styrene (below 20 mM) to (S)-1-phenyl-1,2-ethanediol (two steps in one pot) with 100% conversion. In addition, our CFPS system showed great tolerance to different organic solvents, and, importantly, the entire biocatalytic system can be consistently scaled up without a reduction of the substrate conversion rate. We, therefore, anticipate that our cell-free approach will make a possible cost-effective, high-yielding synthesis of valuable chemicals.

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Cell-free protein synthesis enables one-pot cascade biotransformation in an aqueous-organic biphasic system

Abstract

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Keywords

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