Abstract
Sesquiterpenoids are one of the most diverse classes of isoprenoids which exhibit numerous potentials in industrial biotechnology. The methanotrophs-based methane bioconversion is a promising approach for sustainable production of chemicals and fuels from methane. With intrinsic high carbon flux though the ribulose monophosphate cycle in Methylotuvimicrobium alcaliphilum 20Z, we demonstrated here that employing a short-cut route from ribulose 5-phosphate to 1-deoxy-d-xylulose 5-phosphate (DXP) could enable a more efficient isoprenoid production via the methylerythritol 4-phosphate (MEP) pathway, using α-humulene as a model compound. An additional 2.8-fold increase in α-humulene production yield was achieved by the fusion of the nDXP enzyme and DXP reductase. Additionally, we utilized these engineering strategies for the production of another sesquiterpenoid, α-bisabolene. The synergy of the nDXP and MEP pathways improved the α-bisabolene titer up to 12.24 ± 0.43 mg/gDCW, twofold greater than that of the initial strain. This study expanded the suite of sesquiterpenoids that can be produced from methane and demonstrated the synergistic uses of the nDXP and MEP pathways for improving sesquiterpenoid production in methanotrophic bacteria.
Highlights
IntroductionPublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations
20Z for C5 regeneration, we demonstrated here the potential use of the nDXP pathway for improving the carbon flux through the methylerythritol 4phosphate (MEP) pathway, subsequently enhancing sesquiterpenoid production from methane in M. alcaliphilum 20Z
Alcaliphilum 20Z refined core metabolic pathways of M. alcaliphilum 20Z grown on C1 substrates and indicated that a large portion of carbon flux (~75%) enters the ribulose monophosphate (RuMP) cycle for the regeneration of ribulose-5-phosphate (Ru5P) [13]
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Microorganisms 2021, 9, 1236 methane bioconversion does not compete with food production, which contrasts with other microbial hosts employing biomass-derived substrates, enabling its potential in industrial manufacturing for the sustainable bioproduction of isoprenoids as well as of other biochemicals [9,10]. M. alcaliphilum 20Z, a halophilic methanotroph, was engineered for α-humulene production via the methylerythritol 4-phosphate (MEP) pathway [19]. This demonstrates an important step forward in methanotrophic biocatalysts for the conversion of methane to isoprenoids. 20Z for C5 regeneration, we demonstrated here the potential use of the nDXP pathway for improving the carbon flux through the MEP pathway, subsequently enhancing sesquiterpenoid production from methane in M. alcaliphilum 20Z. This study represents the first demonstration of microbial production of α-bisabolene from methane
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