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

Research output: Contribution to journalArticlepeer-review

4 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 languageEnglish (US)
Pages (from-to)4001-4008
Number of pages8
JournalBiotechnology and Bioengineering
Volume117
Issue number12
DOIs
StatePublished - Dec 2020

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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