Gluconeogenesis in Saccharomyces cerevisiae: determination of fructose-1,6-bisphosphatase activity in cells grown in the presence of glycolytic carbon sources

Foy, J J; Bhattacharjee, J. K.

doi:10.1128/jb.129.2.978-982.1977

Cited by 11 publications

(6 citation statements)

References 19 publications

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“…Additionally, the slower glycolytic rate of galactose did not significantly change ethanol production. This is presumably because the activity of F16BPase was elevated by reduced F26BP level when galactose was used, which can trigger a futile cycle between F6P and F16BP as reported in the previous study (Foy and Bhattacharjee, 1977). The lowered GlcNAc production using galactose as the sole carbon source resembled the effect of PFK-2 disruption (Fig.…”

Section: Introductionsupporting

confidence: 70%

Improved production of N‐acetylglucosamine in Saccharomyces cerevisiae by reducing glycolytic flux

Lee

2016

Biotech & Bioengineering

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Glucosamine and its derivatives are utilized in the food and biomedical industries. However, current production relies on hydrolysis of natural sources, making it difficult to maintain quality and eliminate allergenic risk. Therefore, microbial production with aid of metabolic engineering is required. We previously demonstrated production of N-acetylglucosamine (GlcNAc) in Saccharomyces cerevisiae by overexpressing an allosteric regulation-free Gfa1p mutant and the haloacid dehalogenase-like phosphatase YqaB. In this study, we further improved GlcNAc production by reducing glycolytic flux. Eukaryotic phosphofructokinase 1 (PFK-1) is allosterically activated by fructose 2,6-bisphosphate (F26BP). Disruption of PFK-2, which synthesizes F26BP, resulted in a slight decrease of GlcNAc production and no significant change of glucose consumption and ethanol production. However, when galactose was used as a sole carbon source to the strain without PFK-2, GlcNAc production was significantly increased and ethanol production was reduced, suggesting that further reduction of glycolytic flux can be used to further improve GlcNAc production. The methodology used in this study can be applied to improve production of carbohydrate derivatives in S. cerevisiae. Biotechnol. Bioeng. Biotechnol. Bioeng. 2016;113: 2524-2528. © 2016 Wiley Periodicals, Inc.

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Section: Introductionsupporting

confidence: 70%

Improved production of N‐acetylglucosamine in Saccharomyces cerevisiae by reducing glycolytic flux

Lee

2016

Biotech & Bioengineering

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“…Enzyme activities are expressed as milliunits per milligram of protein at 25°C (1 mU is the enzyme activity that transforms 1 nmol of substrate in 1 min). Statistical analysis and controls for enzyme assays were as previously described (4).…”

Section: Methodsmentioning

confidence: 99%

“…FBPase was determined according to the method of Gancedo et al and Gancedo (7,8). The activity of FBPase was followed by the glucose-6-phosphate dehydrogenase-dependent reduction of nicotinamide adenine dinucleotide phosphate (4).…”

Section: Methodsmentioning

confidence: 99%

“…PFK was determined according to the method of Sols and Salas (28). The activity of PFK was followed by the (x-glycerophosphate dehydrogenase-dependent oxidation of NAD)H (4).…”

Section: Methodsmentioning

confidence: 99%

“…Gancedo et al (6) have reported no FBPase activity in Saccharomvces cereuisiae grown in the presence of 2%k glucose. However, we have observed significant FBPase and PFK activities in cells grown in the presence of 1%glucose, fructose, or galactose (4). These observations prompted us to investigate the precise mode of synthesis and regulation of FBPase and PFK in glucose-and ethanol-grown cells.…”

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confidence: 93%

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Biosynthesis and regulation of fructose-1,6-bisphosphatase and phosphofructokinase in Saccharomyces cerevisiae grown in the presence of glucose and gluconeogenic carbon sources

Foy¹,

Bhattacharjee²

1978

J Bacteriol

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The mode of synthesis and the regulation of fructose-1,6-bisphosphatase (FBPase), a gluconeogenic enzyme, and phosphofructokinase (PFK), a glycolytic enzyme, were investigated in Saccharomyces cerevisiae after growth in the presence of different concentrations of glucose or various gluconeogenic carbon sources. The activity of FBPase appeared in the cells after the complete disappearance of glucose from the growth medium with a concomitant increase of the pH and no significant change in the levels of accumulated ethanol. The appearance of FBPase activity following glucose depletion was dependent upon the synthesis of protein. The FBPase and PFK were present in glucose-, ethanol-, glycerol-, lactate-, or pyruvate-grown cells; however, the time of appearance and the levels of both these enzymes varied. The FBPase activity was always higher in 1% glucose-grown cells than in cells grown in the presence of gluconeogenic carbon sources. Phosphoglucose isomerase activity did not vary significantly. Addition of glucose to an FBPase and PFK synthesizing culture resulted in a complete loss, followed by a reappearance, of FBPase activity and an initial increase, followed by a decrease, of PFK activity. In the presence of cycloheximide the disappearance of glucose and the changes in the levels of FBPase and PFK were decreased significantly. It is concluded that S. cerevisiae exhibits a more efficient synthesis of FBPase after the exhaustion of glucose compared to the activity present in cells grown in the presence of exogenous gluconeogenic carbon sources. Two metabolically antagonistic enzymes, FBPase and PFK, are present during the transition phase, but not during the exponential phase, of growth, and the decay or inactivation of these enzymes in vivo may be dependent upon a glucose-induced protease activity.

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Application of characteristic reaction paths: Rate‐limiting capability of phosphofructokinase in yeast fermentation

Liao

Lightfoot

Jolly

et al. 1988

Biotech & Bioengineering

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The intracellular rate-limiting capability of phosphofructokinase in baker's yeast (Saccharomyces cerevisiae) fermentation is investigated using the reaction path analysis discussed by Liao and Lightfoot in a previous article. It is found that the yeast cells under our experimental conditions indeed exhibit the characteristic behavior, and the characteristic reaction path on the G6P-ATP phase plot is determined for cells fed with different sugars, cells of different strains, and cells in the transition period following rehydration. Results suggest that phosphofructokinase does not limit the CO(2) production rate of the cells under investigation. However, it is not present in a great excess either: ca. 80% of phosphofructokinase activity is utilized by the glycolytic pathway of the cell under investigation.

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Gluconeogenesis in Saccharomyces cerevisiae: determination of fructose-1,6-bisphosphatase activity in cells grown in the presence of glycolytic carbon sources

Cited by 11 publications

References 19 publications

Improved production of N‐acetylglucosamine in Saccharomyces cerevisiae by reducing glycolytic flux

Improved production of N‐acetylglucosamine in Saccharomyces cerevisiae by reducing glycolytic flux

Biosynthesis and regulation of fructose-1,6-bisphosphatase and phosphofructokinase in Saccharomyces cerevisiae grown in the presence of glucose and gluconeogenic carbon sources

Application of characteristic reaction paths: Rate‐limiting capability of phosphofructokinase in yeast fermentation

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