RT Journal Article
SR Electronic
T1 Cell-free protein synthesis enables one-pot cascade biotransformation in an aqueous-organic biphasic system
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.17.995118
DO 10.1101/2020.03.17.995118
A1 Wan-Qiu Liu
A1 Changzhu Wu
A1 Michael C. Jewett
A1 Jian Li
YR 2020
UL http://biorxiv.org/content/early/2020/03/19/2020.03.17.995118.abstract
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 work, 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 reduction of the substrate conversion rate. We therefore anticipate that our cell-free approach will make possible cost-effective, high-yielding synthesis of valuable chemicals.