Whole-cell biotransformation for large scale production of carcinine in Escherichia coli

Zhao, Man; Song, Xiangting; Liu, Wei; Qi, Furong; Zhao, Tingting; Xia, Keke; Liu, Zhiqiang; Zheng, Yu‐Guo

doi:10.1016/j.jbiotec.2022.06.003

Cited by 2 publications

(1 citation statement)

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“…Biotransformation is the enzymatic conversion process utilizing either in vivo engineered microbial whole cells or in vitro purified enzymes. − Due to complete cellular structures and the detrimental effect of toxic intermediates/products on microbial cell growth, the final yield of whole-cell catalysis is often low and unsatisfactory. , In spite of the high efficiency of purified enzyme catalysis, the enzyme purification process is laborious, time-consuming, and expensive, preventing mass production . Recently, cell-free protein synthesis (CFPS) systems have emerged as an alternative that can avoid the disadvantages of the aforementioned two approaches.…”

Section: Introductionmentioning

confidence: 99%

Combining Cell-Free Expression and Multifactor Optimization for Enhanced Biosynthesis of Cinnamyl Alcohol

Zhang

Liu

Ling

et al. 2023

J. Agric. Food Chem.

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Cell-free expression systems have emerged as a potent and promising platform for the biosynthesis of chemicals by reconstituting in vitro expressed enzymes. Here, we report cell-free biosynthesis of cinnamyl alcohol (cinOH) with enhanced productivity by using the Plackett−Burman experimental design for multifactor optimization. Initially, four enzymes were individually expressed in vitro and directly mixed to reconstitute a biosynthetic route for the synthesis of cinOH. Then, the Plackett−Burman experimental design was used to screen multiple reaction factors and found three crucial parameters (i.e., reaction temperature, reaction volume, and carboxylic acid reductase) for the cinOH production. With the optimum reaction conditions, approximately 300 μM of cinOH was synthesized after 10 h of cell-free biosynthesis. Extending the production time to 24 h also increased the production to a maximum yield of 807 μM, which is nearly 10 times higher than the initial yield without optimization. This study demonstrates that cell-free biosynthesis can be combined with other powerful optimization methodologies such as the Plackett−Burman experimental design for enhanced production of valuable chemicals.

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