期刊
METABOLIC ENGINEERING
卷 74, 期 -, 页码 98-107出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2022.10.001
关键词
Metabolic engineering; Enzyme engineering; Model -assisted design; Electrocatalytic reduction; n -Butane
资金
- Suncor Energy Inc
- NSERC CRD grant [OCE-VIP29120]
- Ontario Centres of Excellence VIP Grant
- [CRDPJ 517554-17]
Efforts have been made to develop environmentally friendly alternatives to fossil fuels due to rising concerns about climate change and sustainable energy. This study successfully increased the production of n-butane through enzymatic engineering and model-assisted strain design, paving the way for selectively producing n-butane from renewable carbon sources.
Rising concerns about climate change and sustainable energy have attracted efforts towards developing envi-ronmentally friendly alternatives to fossil fuels. Biosynthesis of n-butane, a highly desirable petro-chemical, fuel additive and diluent in the oil industry, remains a challenge. In this work, we first engineered enzymes Tes, Car and AD in the termination module to improve the selectivity of n-butane biosynthesis, and ancestral recon-struction and a synthetic RBS significantly improved the AD abundance. Next, we did ribosome binding site (RBS) calculation to identify potential metabolic bottlenecks, and then mitigated the bottleneck with RBS en-gineering and precursor propionyl-CoA addition. Furthermore, we employed a model-assisted strain design and a nonrepetitive extra-long sgRNA arrays (ELSAs) and quorum sensing assisted CRISPRi to facilitate a dynamic two -stage fermentation. Through systems engineering, n-butane production was increased by 168-fold from 0.04 to 6.74 mg/L. Finally, the maximum n-butane production from acetate was predicted using parsimonious flux balance analysis (pFBA), and we achieved n-butane production from acetate produced by electrocatalytic CO reduction. Our findings pave the way for selectively producing n-butane from renewable carbon source.
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