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PYC2  -  pyruvate carboxylase 2

Saccharomyces cerevisiae S288c

Synonyms: PCB 2, Pyruvate carboxylase 2, Pyruvic carboxylase 2, YBR1507, YBR218C
 
 
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Disease relevance of PYC2

 

High impact information on PYC2

  • The yeast Pyc1 isoform of pyruvate carboxylase has been further characterized and shown to differ from the Pyc2 isoform in its K(a) for K(+) activation [2].
  • Here we have measured basic kinetic parameters for Pyc1 and found that the K(a) of this isozyme for acetyl CoA is in the order of 8-10-fold higher than previously recorded, suggesting that Pyc1 and Pyc2 may be differentially regulated by this effector [3].
  • Polymorphism of the yeast pyruvate carboxylase 2 gene and protein: effects on protein biotinylation [1].
  • We have confirmed the C-terminus of Pyc2 encoded by the gene isolated here by expressing and purifying an 86-amino-acid biotin-domain peptide [1].
  • On ethanol minimal medium, the growth-related pattern of PYC1 and PYC2 expression was similar as shown by a 3.6-fold decline from early to mid log phase [4].
 

Biological context of PYC2

 

Anatomical context of PYC2

  • The results obtained for CHO cells compared to BHK cells reported previously, indicated that the PYC2 expression dominantly reduced the lactate formation and increased the yield of the recombinant protein to be produced [6].
  • In the present study, a CHO cell line producing recombinant human granulocyte macrophage colony stimulating factor was used to evaluate the impact of PYC2 expression and reduced culture temperature [6].
 

Associations of PYC2 with chemical compounds

  • Yeasts with disruptions in the genes PYC1 and PYC2 encoding the isoenzymes of pyruvate carboxylase cannot grow in a glucose-ammonium medium (Stucka et al. (1991) Mol. Gen. Genet. 229, 307-315) [8].
  • However, only 65% lactate were produced in PYC2-expressing cells and the product yield was 200% higher compared to the control [6].
  • Transfer of exponentially growing cells of the pyc2 mutant from an aspartate or a glutamate medium to an ammonium medium caused a fivefold increase in PYC1 mRNA in less than 30 min, whereas in the inverse experiment, PYC1 transcripts returned within 30 min to the low levels found in aspartate/glutamate medium [9].
 

Analytical, diagnostic and therapeutic context of PYC2

  • Due to the fact that PYC2-expressing cells are characterized by reduced glucose consumption, a prolonged production phase in bioreactors can be maintained [10].

References

  1. Polymorphism of the yeast pyruvate carboxylase 2 gene and protein: effects on protein biotinylation. Val, D.L., Chapman-Smith, A., Walker, M.E., Cronan, J.E., Wallace, J.C. Biochem. J. (1995) [Pubmed]
  2. Kinetic characterization of yeast pyruvate carboxylase isozyme Pyc1 and the Pyc1 mutant, C249A. Branson, J.P., Nezic, M., Jitrapakdee, S., Wallace, J.C., Attwood, P.V. Biochemistry (2004) [Pubmed]
  3. Kinetic characterization of yeast pyruvate carboxylase isozyme pyc1. Branson, J.P., Nezic, M., Wallace, J.C., Attwood, P.V. Biochemistry (2002) [Pubmed]
  4. Regulation of pyruvate carboxylase isozyme (PYC1, PYC2) gene expression in Saccharomyces cerevisiae during fermentative and nonfermentative growth. Brewster, N.K., Val, D.L., Walker, M.E., Wallace, J.C. Arch. Biochem. Biophys. (1994) [Pubmed]
  5. DNA sequences in chromosomes II and VII code for pyruvate carboxylase isoenzymes in Saccharomyces cerevisiae: analysis of pyruvate carboxylase-deficient strains. Stucka, R., Dequin, S., Salmon, J.M., Gancedo, C. Mol. Gen. Genet. (1991) [Pubmed]
  6. Impact of temperature reduction and expression of yeast pyruvate carboxylase on hGM-CSF-producing CHO cells. Fogolín, M.B., Wagner, R., Etcheverrigaray, M., Kratje, R. J. Biotechnol. (2004) [Pubmed]
  7. Regulatory regions in the promoters of the Saccharomyces cerevisiae PYC1 and PYC2 genes encoding isoenzymes of pyruvate carboxylase. Menéndez, J., Gancedo, C. FEMS Microbiol. Lett. (1998) [Pubmed]
  8. A mutation affecting carbon catabolite repression suppresses growth defects in pyruvate carboxylase mutants from Saccharomyces cerevisiae. Blázquez, M.A., Gamo, F.J., Gancedo, C. FEBS Lett. (1995) [Pubmed]
  9. Regulation of pyc1 encoding pyruvate carboxylase isozyme I by nitrogen sources in Saccharomyces cerevisiae. Huet, C., Menendez, J., Gancedo, C., François, J.M. Eur. J. Biochem. (2000) [Pubmed]
  10. Expression of recombinant cytoplasmic yeast pyruvate carboxylase for the improvement of the production of human erythropoietin by recombinant BHK-21 cells. Irani, N., Beccaria, A.J., Wagner, R. J. Biotechnol. (2002) [Pubmed]
 
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