Growth-uncoupled isoprenoid synthesis in Rhodobacter sphaeroides

Biotechnology for Biofuels
Enrico OrsiRuud A Weusthuis

Abstract

Microbial cell factories are usually engineered and employed for cultivations that combine product synthesis with growth. Such a strategy inevitably invests part of the substrate pool towards the generation of biomass and cellular maintenance. Hence, engineering strains for the formation of a specific product under non-growth conditions would allow to reach higher product yields. In this respect, isoprenoid biosynthesis represents an extensively studied example of growth-coupled synthesis with rather unexplored potential for growth-independent production. Rhodobacter sphaeroides is a model bacterium for isoprenoid biosynthesis, either via the native 2-methyl-d-erythritol 4-phosphate (MEP) pathway or the heterologous mevalonate (MVA) pathway, and for poly-β-hydroxybutyrate (PHB) biosynthesis. This study investigates the use of this bacterium for growth-independent production of isoprenoids, with amorpha-4,11-diene as reporter molecule. For this purpose, we employed the recently developed Cas9-based genome editing tool for R. sphaeroides to rapidly construct single and double deletion mutant strains of the MEP and PHB pathways, and we subsequently transformed the strains with the amorphadiene producing plasmid. Furthermore, we em...Continue Reading

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Citations

Nov 6, 2020·Metabolic Engineering·Dongdong ZhaoXueli Zhang
Feb 11, 2021·Journal of Agricultural and Food Chemistry·Jin-Quan HuangXin Fang
Mar 29, 2021·Molecular Biotechnology·Rajveer SinghDipanjan Ghosh

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Methods Mentioned

BETA
genetic modifications
NMR

Software Mentioned

MVA
pBBR

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