Enhancement of pyruvate productivity by inducible expression of a F0F1-ATPase inhibitor INH1 in Torulopsis glabrata CCTCC M202019

Zhou, Jingwen; Huang, Lei; Liu, Li; Chen, Jian

doi:10.1016/j.jbiotec.2009.09.005

Cited by 33 publications

(26 citation statements)

References 39 publications

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“…The modified EGFP expression vector was constructed as follows. The EGFP gene containing four glycine codons in its 5′‐end ( EGFP ( gly4 )) was amplified from the pYES2‐EGFP vector (Zhou, Huang, et al, ) with P11/P12. The EGFP ( gly4 ) product was obtained and digested with XbaI and BamHI, ligated to pET‐20b (+) to obtain pET‐20b‐ EGFP ( gly 4).…”

Section: Methodsmentioning

confidence: 99%

Enhanced pyruvate production in Candida glabrata by carrier engineering

Luo

Liu

et al. 2017

Biotech & Bioengineering

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Pyruvate is an important organic acid that plays a key role in the central metabolic pathway. Manipulating transporters is an efficient strategy to enhance production of target organic acids and a means to understand the effects of altered intracellular pyruvate content on global metabolic networks. Efforts have been made to manipulate mitochondrial pyruvate carrier (MPC) to transport pyruvate into different subcellular compartments in Candida glabrata to demonstrate the effects of the subcellular distribution of pyruvate on central carbon metabolism. By increasing the mitochondrial pyruvate content through enhancing the rate of pyruvate transport into mitochondria, a high central carbon metabolism rate, specific growth rate and specific pyruvate production rate were obtained. Comparing the intracellular pyruvate content of engineered and control strains showed that higher intracellular pyruvate levels were not conducive to improving pyruvate productivity or central carbon metabolism. Plasma membrane expression of MPCs significantly increased the expression levels of key rate-limiting glycolytic enzymes. Moreover, pyruvate production of CGΔura3-Sp-MPC1, CGΔura3-Sp-MPC2, and CGΔura3-Sp-MPC1-Sp-MPC2 increased 134.4%, 120.3%, and 30.0%, respectively. In conclusion, lower intracellular pyruvate content enhanced central carbon metabolism and provided useful clues for improving the production of other organic acids in microorganisms.

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Section: Methodsmentioning

confidence: 99%

Enhanced pyruvate production in Candida glabrata by carrier engineering

Luo

Liu

et al. 2017

Biotech & Bioengineering

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“…A multi‐vitamin auxotrophic C. glabrata CCTCC M202019, screened in our previous study (Zhou et al. 2009a), was used throughout the study.…”

Section: Methodsmentioning

confidence: 99%

“…2009b). Interruption or inhibition of oxidative phosphorylation could significantly decrease the ATP regeneration (Zhou et al. 2009a).…”

Section: Introductionmentioning

confidence: 99%

Improved ATP supply enhances acid tolerance of Candida glabrata during pyruvic acid production

Zhou

Liu

Chen

2010

Journal of Applied Microbiology

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Aims: A major problem in industrial fermentation of organic acids with micro‐organisms is to ensure a suitable pH in the culture broth. To circumvent this problem, we investigated the effect of citrate, which is a widely used auxiliary energy co‐substrate, on cell growth, organic acid production and pH homeostasis among extracellular environment, cytoplasm and vacuole, in the pyruvic acid production by Candida glabrata CCTCC M202019 under different pH conditions. Methods and Results: Analysis of intracellular ATP regeneration, cytoplasmic and vacuolar pH values under different culture conditions points towards a relief of stress when C. glabrata is exposed to lower pH, if citrate is added. When 50 mmol l−1 citrate was added to the culture medium, the intracellular ATP concentrations increased by 20·5% (pH 5·5), 20·4% (pH 5·0) and 39·3% (pH 4·5), and higher pH gradients among the culture broth, cell cytoplasm and vacuoles resulted. As a consequence, the cell growth and pyruvic acid production of C. glabrata CCTCC M202019 were significantly improved under pH 5·0 and 4·5. Conclusions: The acid tolerance of yeast can be improved by enhancing the ATP supply, which helps to maintain higher pH gradients in the system. Significance and Impact of the Study: The results presented here expand our understanding of the physiological characteristics in eukaryotic micro‐organisms under low pH conditions and provide a potential route for the further improvement of organic acids production process by process optimization or metabolic engineering.

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“…More sophisticated methods to control the intracellular ATP content continue to be developed. One example is constructing a copper-inducible F 0 F 1 -ATPase inhibitor, the INH1 gene from S. cerevisiae [53]. When used with a pyruvate-overproducing strain of C. glabrata, this approach increased the volumetric productivity of pyruvate by 23% to 1.69 g/L·h.…”

Section: Metabolic Engineering Of Pathwaysmentioning

confidence: 99%

Recent Progress in the Microbial Production of Pyruvic Acid

Maleki

Eiteman

2017

Fermentation

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Pyruvic acid (pyruvate) is a cellular metabolite found at the biochemical junction of glycolysis and the tricarboxylic acid cycle. Pyruvate is used in food, cosmetics, pharmaceutical and agricultural applications. Microbial production of pyruvate from either yeast or bacteria relies on restricting the natural catabolism of pyruvate, while also limiting the accumulation of the numerous potential by-products. In this review we describe research to improve pyruvate formation which has targeted both strain development and process development. Strain development requires an understanding of carbohydrate metabolism and the many competing enzymes which use pyruvate as a substrate, and it often combines classical mutation/isolation approaches with modern metabolic engineering strategies. Process development requires an understanding of operational modes and their differing effects on microbial growth and product formation.

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Enhancement of pyruvate productivity by inducible expression of a F0F1-ATPase inhibitor INH1 in Torulopsis glabrata CCTCC M202019

Cited by 33 publications

References 39 publications

Enhanced pyruvate production in Candida glabrata by carrier engineering

Enhanced pyruvate production in Candida glabrata by carrier engineering

Improved ATP supply enhances acid tolerance of Candida glabrata during pyruvic acid production

Recent Progress in the Microbial Production of Pyruvic Acid

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