Metabolic Redesign of Rhodobacter sphaeroides for Lycopene Production

Su, Anping; Chi, Shuang; Liu, Ying; Tan, Siyuan; Shan, Qiang; Chen, Zhi; Meng, Yan

doi:10.1021/acs.jafc.8b00855

Cited by 61 publications

(78 citation statements)

References 42 publications

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“…The Actin gene was used as the internal control; the Primer and Sequence used during q-PCR see supplementary Table S1. The relative gene expression analysis was performed using the method published previously 41 .…”

Section: Methodsmentioning

confidence: 99%

Dissolved-oxygen feedback control fermentation for enhancing β-carotene in engineered Yarrowia lipolytica

Shan

Liu

et al. 2020

Sci Rep

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The DO-stat fed-batch fermentation was carried out to explore the volumetric productivity of β-carotene in engineered Yarrowia lipolytica C11 strain. Using DO-stat fed-batch fermentation, we achieved 94 g/L biomass and 2.01 g/L β-carotene. Both biomass and β-carotene were about 1.28-fold higher than that in fed-batch fermentation. The ATP, NADP+/NADPH, and gene expression levels of tHMG, GGS1, carRA, and carB were promoted as compared to that in fed-batch fermentation. As for as the kinetic parameters in DO-stat fed-batch fermentation, μm′, Yx/s′, and Yp/s′ was 0.527, 0.353, and 0.158, respectively. The μm′ was elevated 4.66-fold than that in fed-batch fermentation. These data illustrate that more dissolved oxygen increased the biomass. The Yx/s′ and Yp/s′ were increased 1.15 and 22.57-fold, which suggest that the DO-stat fed-batch fermentation reduced the Crabtree effect and improved the utilization rate of glucose. Therefore, DO-stat fed-batch fermentation is a promising strategy in the industrialized production of β-carotene.

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Section: Methodsmentioning

confidence: 99%

Dissolved-oxygen feedback control fermentation for enhancing β-carotene in engineered Yarrowia lipolytica

Shan

Liu

et al. 2020

Sci Rep

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“…Moreover, it is also a natural producer of coenzyme Q 10 . Apart from these native isoprenoid molecules, heterologous production of lycopene [27] and sesquiterpenes [28] has also been reported in this species.…”

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confidence: 93%

Growth-uncoupled isoprenoid synthesis in Rhodobacter sphaeroides

Orsi

Mougiakos

Post

et al. 2020

Biotechnol Biofuels

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Background: 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. Results: 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 employed 13 C-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. Conclusions: Apart 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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“…As a safe and non-toxic photosynthetic bacteria, R. sphaeroides has been widely studied in its unique biological transformation functions [23,24,25]. Besides, R. sphaeroides serves as a new platform for producing bioactive substance including coenzyme Q10, carotenoids, sesquiterpene, fatty acids, and 5-aminolevulinic acid [26,27,28,29,30]. Owing to its excellent adaptability to the aerobic or anaerobic environment and fermentation, or photosynthesis conditions, R. sphaeroides has attracted considerable attention in food, breeding, environmental, and pharmaceutical industries [22,24,31,32].…”

Section: Introductionmentioning

confidence: 99%

The Effects of Rhodobacter sphaeroides on the Composition of Gut Microbiota and Short-chain Fatty Acids in Mice

Yang¹,

Luan²,

An³

et al. 2020

JFNR

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This study aims to determine the effects of Rhodobacter sphaeroides 8513 on the physiological and biochemical indicators, short-chain fatty acids, and gut microbiota of healthy BALB/c mice. Mice were divided into the control group and R. sphaeroides group (5×10 8 CFU/mL). After 28 days of continuous gavage, fecal samples were collected for high-throughput sequencing, then mice were sacrificed and the serum was taken for biochemical analysis. Liver, kidney, and spleen were obtained for organ coefficient calculation, and cecum contents were collected for short-chain fatty acid analysis by gas chromatography. The results showed that there was no significant difference in body weight gain, alanine aminotransferase (ALT) and urea nitrogen (BUN) levels between the control group and R. sphaeroides group, while R. sphaeroides significantly decreased the aspartate aminotransferase (AST) and creatinine (CRE) level. No side effect of R. sphaeroides treatment on the basic physiological health of mice was observed. R. sphaeroides significantly increased the content of acetic acids in the gut of mice, while there were no significant changes in the concentration of butyric and propionic acid. High-throughput sequencing analysis indicated that there was a significant difference in α-diversity between these 2 groups, while there was no significant difference in β-diversity between them. R. sphaeroides not only increased the abundance of anaerobic bacteria of Rhodospirillaceae, Desulfovibrionaceae, and Helicobacter in the gut of mice, but also increased the abundance of the predominant microflora in the gut such as Bacteroidaceae (Bacteroides), Prevotellaceae, Clostridiales_vadinBB60_group (Closporaceae). These results suggested that R. sphaeroides 8513 could modulate the microbial ecology in the gut of mice.

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Metabolic Redesign of Rhodobacter sphaeroides for Lycopene Production

Cited by 61 publications

References 42 publications

Dissolved-oxygen feedback control fermentation for enhancing β-carotene in engineered Yarrowia lipolytica

Dissolved-oxygen feedback control fermentation for enhancing β-carotene in engineered Yarrowia lipolytica

Growth-uncoupled isoprenoid synthesis in Rhodobacter sphaeroides

The Effects of <i>Rhodobacter sphaeroides</i> on the Composition of Gut Microbiota<i> </i>and Short-chain Fatty Acids in Mice

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