Systems engineering of Escherichia coli for n-butane production
Allbwn ymchwil: Cyfraniad at gyfnodolyn › Erthygl › adolygiad gan gymheiriaid
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Yn: Metabolic Engineering, Cyfrol 74, 01.11.2022, t. 98-107.
Allbwn ymchwil: Cyfraniad at gyfnodolyn › Erthygl › adolygiad gan gymheiriaid
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T1 - Systems engineering of Escherichia coli for n-butane production
AU - Liu, Yilan
AU - Khusnutdinova, Anna
AU - Chen, Jinjin
AU - Crisante, David
AU - Batyrova, Khorcheska
AU - Raj, Kaushik
AU - Feigis, Michelle
AU - Shirzadi, Erfan
AU - Wang, Xiaotong
AU - Dorakhan, Roham
AU - Wang, Xue
AU - Stogios, Peter J
AU - Yakunin, Alexander F
AU - Sargent, Edward H
AU - Mahadevan, Radhakrishnan
N1 - Crown Copyright © 2022. Published by Elsevier Inc. All rights reserved.
PY - 2022/11/1
Y1 - 2022/11/1
N2 - Rising concerns about climate change and sustainable energy have attracted efforts towards developing environmentally 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 reconstruction 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 engineering 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.
AB - Rising concerns about climate change and sustainable energy have attracted efforts towards developing environmentally 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 reconstruction 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 engineering 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.
KW - Escherichia coli/genetics
KW - Metabolic Engineering
KW - Butanes/metabolism
KW - Acetates/metabolism
U2 - 10.1016/j.ymben.2022.10.001
DO - 10.1016/j.ymben.2022.10.001
M3 - Article
C2 - 36244545
VL - 74
SP - 98
EP - 107
JO - Metabolic Engineering
JF - Metabolic Engineering
SN - 1096-7176
ER -