The isolation and characterization of the pyruvate kinase-encoding gene from the yeast Yarrowia lipolytica

Strick, Christine A.; James, Larry C.; O'Donnell, M.M.; Gollaher, M.G.; Franke, Arthur E.

doi:10.1016/0378-1119(94)90746-3

“…There are no reports of the existence of more than one pyruvate kinase encoding gene in NCY. Only the genes encoding pyruvate kinases from S. pombe and Y. lipolytica have been cloned [89,90]. Overexpression of the pyk1 gene from S. pombe has allowed puri¢cation of the enzyme that behaves as a dimer [89].…”

Section: From Trioses Phosphate To Pyruvatementioning

confidence: 99%

“…The organization of the YlPYK1 gene from Y. lipolytica is curious as it presents two introns separated by a short stretch of 45 bp. [90,91] In general, pyruvate kinase from diverse origins presents a sigmoidal kinetics with respect to PEP; fructose-1,6-bisphosphate activates the enzyme and converts its kinetic to an hyperbolic one. In contrast, the enzyme from C. utilis [92] and Y. lipolytica [93] are not activated by fructose-1,6bisphosphate and in this last yeast the kinetics towards PEP is hyperbolic [93].…”

Section: From Trioses Phosphate To Pyruvatementioning

confidence: 99%

Carbohydrate and energy-yielding metabolism in non-conventional yeasts

Flores¹

2000

FEMS Microbiology Reviews

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Sugars are excellent carbon sources for all yeasts. Since a vast amount of information is available on the components of the pathways of sugar utilization in Saccharomyces cerevisiae it has been tacitly assumed that other yeasts use sugars in the same way. However, although the pathways of sugar utilization follow the same theme in all yeasts, important biochemical and genetic variations on it exist. Basically, in most non-conventional yeasts, in contrast to S. cerevisiae, respiration in the presence of oxygen is prominent for the use of sugars. This review provides comparative information on the different steps of the fundamental pathways of sugar utilization in non-conventional yeasts: glycolysis, fermentation, tricarboxylic acid cycle, pentose phosphate pathway and respiration. We consider also gluconeogenesis and, briefly, catabolite repression. We have centered our attention in the genera Kluyveromyces, Candida, Pichia, Yarrowia and Schizosaccharomyces, although occasional reference to other genera is made. The review shows that basic knowledge is missing on many components of these pathways and also that studies on regulation of critical steps are scarce. Information on these points would be important to generate genetically engineered yeast strains for certain industrial uses.

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“…There are no reports of the existence of more than one pyruvate kinase encoding gene in NCY. Only the genes encoding pyruvate kinases from S. pombe and Y. lipolytica have been cloned [89,90]. Overexpression of the pyk1 + gene from S. pombe has allowed purification of the enzyme that behaves as a dimer [89].…”

Section: The Common Glycolytic Trunkmentioning

confidence: 99%

“…The organization of the YlPYK1 gene from Y. lipolytica is curious as it presents two introns separated by a short stretch of 45 bp. [90,91]…”

Section: The Common Glycolytic Trunkmentioning

confidence: 99%

Carbohydrate and energy-yielding metabolism in non-conventional yeasts: Figure 1

Flores

¹

,

Rodrı́guez

²

,

Petit

³

et al. 2000

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Sugars are excellent carbon sources for all yeasts. Since a vast amount of information is available on the components of the pathways of sugar utilization in Saccharomyces cerevisiae it has been tacitly assumed that other yeasts use sugars in the same way. However, although the pathways of sugar utilization follow the same theme in all yeasts, important biochemical and genetic variations on it exist. Basically, in most non-conventional yeasts, in contrast to S. cerevisiae, respiration in the presence of oxygen is prominent for the use of sugars. This review provides comparative information on the different steps of the fundamental pathways of sugar utilization in non-conventional yeasts: glycolysis, fermentation, tricarboxylic acid cycle, pentose phosphate pathway and respiration. We consider also gluconeogenesis and, briefly, catabolite repression. We have centered our attention in the genera Kluyveromyces, Candida, Pichia, Yarrowia and Schizosaccharomyces, although occasional reference to other genera is made. The review shows that basic knowledge is missing on many components of these pathways and also that studies on regulation of critical steps are scarce. Information on these points would be important to generate genetically engineered yeast strains for certain industrial uses.

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“…The N-terminal region upstream from the start of the homologous subtilisin domain is over 130 amino acids larger in Xpr6p than in Kex2p and Kexlp. This larger size is most likely not due to an intron as neither the stringent consensus mRNA splice site sequences and TACTAAC box of S. cerevisiae nor the less strictly conserved consensus sequences from Schizosaccharomyces pombe, Neurospora crassa (Gallwitz et al, 1987) or Y. lipolytica (Strick et al, 1992) are found in this region.…”

Section: S M E a D K R D N D R Q N L L G * * * Tccatggaggctgac Aagcggmentioning

confidence: 99%

Cloning, nucleotide sequence and functions of XPR6, which codes for a dibasic processing endoprotease from the yeast Yarrowia lipolytica

Enderlin

¹

,

Ogrydziak

²

1994

Yeast

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Yarrowia lipolytica DO613, carrying the xpr6-13 mutation, secretes an inactive precursor of alkaline extracellular protease that has not been cleaved after the Lys-Arg at the end of the pro-region. Compared to wild type, DO613 membrane preparations had significantly reduced ability to cleave after Lys-Arg of an artificial substrate. The XPR6 gene was cloned by complementation by screening for restoration of production of alkaline protease activity. Sequencing of a 3735 base pair SalI-SphI XPR6 fragment revealed a large open reading frame with a coding capacity of 976 amino acids (molecular weight, 110,016). The deduced amino acid sequence had significant homology to Saccharomyces cerevisiae Kex2p, a processing endoprotease that cleaves after pairs of basic amino acids. Disruption of the XPR6 gene was not lethal, but it resulted in several phenotypic changes. First, essentially no mature alkaline extracellular protease was produced indicating that the low levels produced by strains carrying previously isolated xpr6 alleles were due to leaky mutations. Second, mating type B strains carrying the disrupted XPR6 gene did not mate, but mating type A strains did. Third, the XPR6 disruption strains grew poorly on rich media at pH 5.5 and above. Cells remained physically attached after budding and continued to bud forming large dog balloon-like structures. In addition, these structures aggregated forming visible clumps in liquid culture. These growth aberrations were largely eliminated by growing cells in medium at pH 4. Fourth, no mycelial forms were observed regardless of the pH.

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The isolation and characterization of the pyruvate kinase-encoding gene from the yeast Yarrowia lipolytica

Cited by 7 publications

References 3 publications

Carbohydrate and energy-yielding metabolism in non-conventional yeasts

Carbohydrate and energy-yielding metabolism in non-conventional yeasts

Carbohydrate and energy-yielding metabolism in non-conventional yeasts: Figure 1

Cloning, nucleotide sequence and functions of XPR6, which codes for a dibasic processing endoprotease from the yeast Yarrowia lipolytica

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