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VITAMIN A AND ISOPRENOID SYNTHESIS IN THE RAT.

ABSTRACT: 1. Vitamin A-deficient rats were compared with similar animals given small amounts of vitamin A sufficient for adequate growth and with animals given large amounts of vitamin A. The effects of pair-feeding and feeding ad libitum were compared. 2. Ubiquinone and cholesterol concentrations in liver were measured at various stages of the deficiency, and the uptake of radioactive mevalonate and acetate into isoprenoid compounds was studied. 3. Ubiquinone concentrations in liver increased markedly in deficient rats compared with adequate controls, and heavy vitamin A supplementation had a further effect in depressing ubiquinone concentrations. These effects were unrelated to food intake or to the size of the organs. 4. Radioactive uptake into ubiquinone was often greater in deficient livers, especially during the early stages of the experiments, but the effect was not consistent. 5. Cholesterol concentrations were usually higher in deficient livers and these were more affected by the feeding regimen. 6. No consistent effect of vitamin A deficiency or of vitamin A dosage on the incorporation of mevalonate into cholesterol or squalene was found. 7. No evidence has been found for a specific effect of vitamin A on isoprenoid synthesis at the metabolic level.

SUBMITTER: DIPLOCK AT

PROVIDER: S-EPMC1215186 | biostudies-other | 1965 Apr

REPOSITORIES: biostudies-other

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Project description:BackgroundMicrobial 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.ResultsThis 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 employed 13C-metabolic flux ratio analysis to monitor the changes in the isoprenoid metabolic fluxes under different cultivation conditions. We demonstrated that active flux via both isoprenoid pathways while inactivating PHB synthesis maximizes growth-coupled isoprenoid synthesis. On the other hand, the strain that showed the highest growth-independent isoprenoid yield and productivity, combined the plasmid-based heterologous expression of the orthogonal MVA pathway with the inactivation of the native MEP and PHB production pathways.ConclusionsApart from proposing a microbial cell factory for growth-independent isoprenoid synthesis, this work provides novel insights about the interaction of MEP and MVA pathways under different growth conditions.

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VITAMIN A AND ISOPRENOID SYNTHESIS IN THE RAT.

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