Factors Affecting Glycerolproduction by a Bioconversion Process with a Triose Phosphate Isomerase Deficient Mutant of Saccharomyces Cerevisiae

Compagno, Concetta; Boschi, Francesco; Ranzi, Bianca Maria

doi:10.3109/10242429809003617

Cited by 9 publications

(4 citation statements)

References 9 publications

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“…The highest glycerol yield and productivity reported to date for metabolically engineered S. cerevisiae were observed with a tpi1⌬ deletion mutant (9,10) (Table 1). Apparently, in tpi1⌬ mutants, which lack the glycolytic enzyme triose phosphate isomerase, accumulation of DHAP is prevented by its conversion to glycerol (Fig.…”

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Metabolic Engineering of Glycerol Production inSaccharomyces cerevisiae

Overkamp

Bakker

Kötter

et al. 2002

Appl Environ Microbiol

110

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Inactivation of TPI1, the Saccharomyces cerevisiae structural gene encoding triose phosphate isomerase, completely eliminates growth on glucose as the sole carbon source. In tpi1-null mutants, intracellular accumulation of dihydroxyacetone phosphate might be prevented if the cytosolic NADH generated in glycolysis by glyceraldehyde-3-phosphate dehydrogenase were quantitatively used to reduce dihydroxyacetone phosphate to glycerol. We hypothesize that the growth defect of tpi1-null mutants is caused by mitochondrial reoxidation of cytosolic NADH, thus rendering it unavailable for dihydroxyacetone-phosphate reduction. To test this hypothesis, a tpi1⌬ nde1⌬ nde2⌬ gut2⌬ quadruple mutant was constructed. NDE1 and NDE2 encode isoenzymes of mitochondrial external NADH dehydrogenase; GUT2 encodes a key enzyme of the glycerol-3-phosphate shuttle. It has recently been demonstrated that these two systems are primarily responsible for mitochondrial oxidation of cytosolic NADH in S. cerevisiae. Consistent with the hypothesis, the quadruple mutant grew on glucose as the sole carbon source. The growth on glucose, which was accompanied by glycerol production, was inhibited at high-glucose concentrations. This inhibition was attributed to glucose repression of respiratory enzymes as, in the quadruple mutant, respiratory pyruvate dissimilation is essential for ATP synthesis and growth. Serial transfer of the quadruple mutant on high-glucose media yielded a spontaneous mutant with much higher specific growth rates in high-glucose media (up to 0.10 h ؊1 at 100 g of glucose · liter ؊1 ). In aerated batch cultures grown on 400 g of glucose · liter ؊1 , this engineered S. cerevisiae strain produced over 200 g of glycerol · liter ؊1 , corresponding to a molar yield of glycerol on glucose close to unity.

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“…However, S. cerevisiae tpi1⌬ mutants are unable to grow on glucose as the sole carbon source (9-11). Therefore, biomass was pregrown on glucose-ethanol mixtures, followed by a bioconversion of glucose to glycerol (9,10). During the bioconversion phase, glycerol productivity decreased strongly with time ( Table 1).…”

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confidence: 99%

Metabolic Engineering of Glycerol Production inSaccharomyces cerevisiae

Overkamp

Bakker

Kötter

et al. 2002

Appl Environ Microbiol

110

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“…In our laboratory, we previously developed different S. cerevisiae strains deleted in the TPI1 gene (⌬TPI1), coding for TIM. We used these mutants to obtain elevated productions of glycerol (i.e., 80 g/liter) with high yield on the carbon source (molar ratio, 80 to 90%) (5,6). With the aim to com-pare the physiological relevance of TIM and glycerol production in the Crabtree-positive S. cerevisiae and in the Crabtreenegative K. lactis yeast strains, we decided to characterize the TPI1 gene from K. lactis.…”

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confidence: 99%

“…Glycerol production by wild-type and mutant strains. By means of simple bioconversion processes, S. cerevisiae ⌬TPI1 mutants can be used to produce glycerol with high yield (5,6).…”

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Isolation, Nucleotide Sequence, and Physiological Relevance of the Gene Encoding Triose Phosphate Isomerase from Kluyveromyces lactis

Compagno

Boschi

Daleffe

et al. 1999

Appl Environ Microbiol

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Lack of triose phosphate isomerase activity (TIM) is of special interest because this enzyme works at an important branch point of glycolytic flux. In this paper, we report the cloning and sequencing of the Kluyveromyces lactis gene encoding TIM. UnlikeSaccharomyces cerevisiae ΔTPI1 mutants, the K. lactis mutant strain was found to be able to grow on glucose. Preliminary bioconversion experiments indicated that, like the S. cerevisiae TIM-deficient strain, the K. lactisTIM-deficient strain is able to produce glycerol with high yield.

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Current Awareness

2001

Yeast

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (4 weeks journals ‐ search completed 20th Sept. 2000)

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Factors Affecting Glycerolproduction by a Bioconversion Process with a Triose Phosphate Isomerase Deficient Mutant of Saccharomyces Cerevisiae

Cited by 9 publications

References 9 publications

Metabolic Engineering of Glycerol Production inSaccharomyces cerevisiae

Metabolic Engineering of Glycerol Production inSaccharomyces cerevisiae

Isolation, Nucleotide Sequence, and Physiological Relevance of the Gene Encoding Triose Phosphate Isomerase from Kluyveromyces lactis

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