Design of a synthetic enzyme cascade for the in vitro fixation of formaldehyde to acetoin

Zhenzhen Cui; Mengnan Ding; Wei Dai; Meiyu Zheng; Zhiwen Wang; Tao Chen

doi:10.1016/j.enzmictec.2024.110446

Design of a synthetic enzyme cascade for the in vitro fixation of formaldehyde to acetoin

Enzyme Microb Technol. 2024 Apr 10:178:110446. doi: 10.1016/j.enzmictec.2024.110446. Online ahead of print.

Authors

Zhenzhen Cui¹, Mengnan Ding¹, Wei Dai¹, Meiyu Zheng¹, Zhiwen Wang¹, Tao Chen²

Affiliations

¹ Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, China; Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, China.
² Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, China; Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, China. Electronic address: chentao@tju.edu.cn.

PMID: 38626535
DOI: 10.1016/j.enzmictec.2024.110446

Abstract

Formaldehyde (FALD) has gained prominence as an essential C1 building block in the synthesis of valuable chemicals. However, there are still challenges in converting FALD into commodities. Recently, cell-free biocatalysis has emerged as a popular approach for producing such commodities. Acetoin, also known as 3-hydroxy-2-butanone, has been widely used in food, cosmetic, agricultural and the chemical industry. It is valuable to develop a process to produce acetoin from FALD. In this study, a cell-free multi-enzyme catalytic system for the production of acetoin using FALD as the substrate was designed and constructed. It included three scales: FALD utilization pathway, glycolysis pathway and acetoin synthesis pathway. After the optimization of the reaction system, 20.17 mM acetoin was produced from 122 mM FALD, with a yield of 0.165 mol/mol, reaching 99.0% of the theoretical yield. The pathway provides a new approach for high-yield acetoin production from FALD, which consolidates the foundation for the production of high value-added chemicals using cheap one-carbon compounds.

Keywords: Acetoin; Formaldehyde; In vitro enzyme cascade.