Engineering Vibrio alginolyticus as a novel chassis for PHB production from starch

Li, Hongfei; Tian, Linyue; Lian, Guoli; Fan, Li; Li, Zhengjun

doi:10.3389/fbioe.2023.1130368

Cited by 5 publications

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References 63 publications

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“…Since the construction of E. coli strains that can grow with methanol as the sole carbon source is still difficult [ 9 ], in this study, the PHB biosynthesis from methanol was conducted via whole-cell biocatalysis ( Figure 4 ). Although the final PHB content from methanol (6.19%) was not as high as those previously reported (ranging from 20% to 29% PHB content) from sugar-based carbon sources [ 26 ] or other carbon sources [ 41 , 42 , 43 ], a significant increase of 65% in PHB content with the MCC pathway was observed relative to that with only the RuMP pathway through statistical analysis. The experimental results confirm the importance of the combination of NOG with RuMP to form the complete MCC pathway, which can obviously facilitate the biosynthesis of acetyl-CoA–derived chemicals from one-carbon compounds.…”

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confidence: 64%

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

et al. 2023

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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.

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