Back to Search Start Over

Engineering Escherichia coli for Poly-β-hydroxybutyrate Production from Methanol

Authors :
Jiaying Wang
Zhiqiang Chen
Xiaogui Deng
Qianqian Yuan
Hongwu Ma
Source :
Bioengineering, Vol 10, Iss 4, p 415 (2023)
Publication Year :
2023
Publisher :
MDPI AG, 2023.

Abstract

The naturally occurring one-carbon assimilation pathways for the production of acetyl-CoA and its derivatives often have low product yields because of carbon loss as CO2. We constructed a methanol assimilation pathway to produce poly-3-hydroxybutyrate (P3HB) using the MCC pathway, which included the ribulose monophosphate (RuMP) pathway for methanol assimilation and non-oxidative glycolysis (NOG) for acetyl-CoA (precursor for PHB synthesis) production. The theoretical product carbon yield of the new pathway is 100%, hence no carbon loss. We constructed this pathway in E. coli JM109 by introducing methanol dehydrogenase (Mdh), a fused Hps–phi (hexulose-6-phosphate synthase and 3-phospho-6-hexuloisomerase), phosphoketolase, and the genes for PHB synthesis. We also knocked out the frmA gene (encoding formaldehyde dehydrogenase) to prevent the dehydrogenation of formaldehyde to formate. Mdh is the primary rate-limiting enzyme in methanol uptake; thus, we compared the activities of three Mdhs in vitro and in vivo and then selected the one from Bacillus methanolicus MGA3 for further study. Experimental results indicate that, in agreement with the computational analysis results, the introduction of the NOG pathway is essential for improving PHB production (65% increase in PHB concentration, up to 6.19% of dry cell weight). We demonstrated that PHB can be produced from methanol via metabolic engineering, which provides the foundation for the future large-scale use of one-carbon compounds for biopolymer production.

Details

Language :
English
ISSN :
23065354
Volume :
10
Issue :
4
Database :
Directory of Open Access Journals
Journal :
Bioengineering
Publication Type :
Academic Journal
Accession number :
edsdoj.7690fdd35f9943c2b7e91d8241ab7ed1
Document Type :
article
Full Text :
https://doi.org/10.3390/bioengineering10040415