Improvement of Coenzyme Q10Production by Increasing the NADH/NAD+Ratio inAgrobacterium tumefaciens

Koo, Bong Seong; Gong, Yun Jung; Kim, Sang Yong; Kim, Chan Wha; Lee, Hyeon-Cheol

doi:10.1271/bbb.100034

Cited by 18 publications

(9 citation statements)

References 17 publications

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“…In this study, we obtained a mutant with CoQ 10 productivity of 7.16 mg/g DCW from 12 mutants. It was twice folds as high as the wild-type and also higher than other microbes, which were reported to produce CoQ 10 no more than 3.63 mg/g DCW in shake-flask experiments (Choi et al, 2009;Kim et al, 2006;Koo et al, 2010;Park et al, 2005;Zahiri et al, 2006;Zhang et al, 2007). It showed that with a set of RBS sequences and a pair of promoter and the selfregulator, we can simplify the screening process to obtain the desirable phenotype.…”

Section: Discussionmentioning

confidence: 91%

Enhanced production of coenzyme Q₁₀ by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

et al. 2013

Biotech & Bioengineering

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Fine-tuning the expression level of an engineered pathway is crucial for the metabolic engineering of a host toward a desired phenotype. However, most engineered hosts suffer from nonfunctional protein expression, metabolic imbalance, cellular burden or toxicity from intermediates when an engineered pathway is first introduced, which can decrease production of the desired product. To circumvent these obstacles, we developed a self-regulation system utilizing the trc/tac promoter, LacI(q) protein and ribosomal binding sites (RBS). With the purpose of improving coenzyme Q10 (CoQ10 ) production by increasing the decaprenyl diphosphate supplement, enzymes DXS, DXR, IDI, and IspD were constitutively overexpressed under the control of the trc promoter in Rhodobacter sphaeroides. Then, a self-regulation system combining a set of RBSs for adjusting the expression of the LacI(q) protein was applied to tune the expression of the four genes, resulting in improved CoQ10 production. Finally, another copy of the tac promoter with the UbiG gene (involved in the ubiquinone pathway of CoQ10 biosynthesis) was introduced into the engineered pathway. By optimizing the expression level of both the upstream and downstream pathway, CoQ10 production in the mutants was improved up to 93.34 mg/L (7.16 mg/g DCW), about twofold of the wild-type (48.25 mg/L, 3.24 mg/g DCW).

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Section: Discussionmentioning

confidence: 91%

Enhanced production of coenzyme Q₁₀ by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

et al. 2013

Biotech & Bioengineering

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“…Additionally, we found a higher NADH‐to‐NAD ratio in the NaAc supplemented culture (3.2 ± 1.5 at 19 hr) compared to the control (0.82 at 24 hr) (Figure c). This ratio level (∼3) may seem high but such high ratio levels have been reported in literature for various bacteria with 2.2 for B. subtilis (Gyan, Shiohira, Sato, Takeuchi, & Sato, ), and with 1.6 in Agrobacterium tumefaciens (Koo, Gong, Kim, Kim, & Lee, ). However, the NADPH‐to‐NADP ratio shows being stable and at the same level in both the test and the control cultures, which suggests the ratio robustness.…”

Section: Resultsmentioning

confidence: 81%

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

Zhao

Kasbi

Chen

et al. 2017

Biotech & Bioengineering

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The present study reveals that supplementing sodium acetate (NaAc) strongly stimulates riboflavin production in acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum ATCC 824 with xylose as carbon source. Riboflavin production increased from undetectable concentrations to ∼0.2 g L (0.53 mM) when supplementing 60 mM NaAc. Of interest, solvents production and biomass yield were also promoted with fivefold acetone, 2.6-fold butanol, and 2.4-fold biomass adding NaAc. A kinetic metabolic model, developed to simulate ABE biosystem, with riboflavin production, revealed from a dynamic metabolic flux analysis (dMFA) simultaneous increase of riboflavin (ribA) and GTP (precursor of riboflavin) (PurM) synthesis flux rates under NaAc supplementation. The model includes 23 fluxes, 24 metabolites, and 72 kinetic parameters. It also suggested that NaAc condition has first stimulated the accumulation of intracellular metabolite intermediates during the acidogenic phase, which have then fed the solventogenic phase leading to increased ABE production. In addition, NaAc resulted in higher intracellular levels of NADH during the whole culture. Moreover, lower GTP-to-adenosine phosphates (ATP, ADP, AMP) ratio under NaAc supplemented condition suggests that GTP may have a minor role in the cell energetic metabolism compared to its contribution to riboflavin synthesis.

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“…[15] Ha et al [16] described the optimization of culture conditions like pH and temperature as being critical for enhanced CoQ10 production from Agrobacterium tumefaciens. Metabolic engineering strategies like the introduction of heterologous genes involved in the mevalonate pathway, [18] varying the NADH=NAD þ ratio, [35] limited dissolved oxygen supply, [37] and optimization of feeding strategies for decaprenyl diphosphate synthase gene from A. tumefaciens in Escherichia coli produced higher CoQ10 than wild-type strains. Despite a lower CoQ10 production than the aforesaid and earlier reports, [33,37,38] this study was first of its kind to report CoQ10 production by the less exploited red yeast R. glutinis using better optimized feeding strategies.…”

Section: Modeling Process Parameters For Production Of Coq10mentioning

confidence: 99%

Modeling of Process Parameters for Enhanced Production of Coenzyme Q10 FromRhodotorula glutinis

Balakumaran

Meenakshisundaram

2014

Preparative Biochemistry and Biotechnology

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Coenzyme Q10 (CoQ10) plays an indispensable role in ATP generation through oxidative phosphorylation and helps in scavenging superoxides generated during electron transfer reactions. It finds extensive applications specifically related to oxidative damage and metabolic dysfunctions. This article reports the use of a statistical approach to optimize the concentration of key variables for the enhanced production of CoQ10 by Rhodotorula glutinis in a lab-scale fermenter. The culture conditions that promote optimum growth and CoQ10 production were optimized and the interaction of significant variables para-hydroxybenzoic acid (PHB, 819.34 mg/L) and soybean oil (7.78% [v/v]) was studied using response surface methodology (RSM). CoQ10 production increased considerably from 10 mg/L (in control) to 39.2 mg/L in batch mode with RSM-optimized precursor concentration. In the fed-batch mode, PHB and soybean oil feeding strategy enhanced CoQ10 production to 78.2 mg/L.

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Improvement of Coenzyme Q₁₀Production by Increasing the NADH/NAD⁺Ratio inAgrobacterium tumefaciens

Cited by 18 publications

References 17 publications

Enhanced production of coenzyme Q₁₀ by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

Enhanced production of coenzyme Q₁₀ by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

Modeling of Process Parameters for Enhanced Production of Coenzyme Q10 FromRhodotorula glutinis

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Improvement of Coenzyme Q10Production by Increasing the NADH/NAD+Ratio inAgrobacterium tumefaciens

Cited by 18 publications

References 17 publications

Enhanced production of coenzyme Q10 by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

Enhanced production of coenzyme Q10 by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

Modeling of Process Parameters for Enhanced Production of Coenzyme Q10 FromRhodotorula glutinis

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Improvement of Coenzyme Q₁₀Production by Increasing the NADH/NAD⁺Ratio inAgrobacterium tumefaciens

Enhanced production of coenzyme Q₁₀ by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides

Enhanced production of coenzyme Q₁₀ by self‐regulating the engineered MEP pathway in Rhodobacter sphaeroides