Inorganic polyphosphate is involved in architecture and functioning of yeast cell wall. The strain of Saccharomyces cerevisiae constitutively overexpressing acid phosphatase Pho5 was constructed for studying the Pho5 properties and its possible participation in polyphosphate metabolism. The parent strain was transformed by the vector carrying the PHO5 gene under a strong constitutive promoter of glyceraldehyde-3-phosphate dehydrogenase of S. cerevisiae. The culture liquid and biomass of transformant strain contained approximately equal total acid phosphatase activity. The levels of acid phosphatase activity associated with the cell wall and culture liquid increased in the transformant strain compared to the parent strain ~ 10- and 20-fold, respectively. The Pho5 preparation (specific activity of 46 U/mg protein and yield of 95 U/L) was obtained from culture liquid of overproducing strain. The overproducing strain had no changes in polyphosphate level. The activity of Pho5 with long-chained polyP was negligible. We concluded that Pho5 is not involved in polyphosphate metabolism. Purified Pho5 showed a similar activity with p-nitrophenylphosphate, ATP, ADP, glycerophosphate, and glucose-6-phosphate. The substrate specificity of Pho5 and its extracellular localization suggest its function: the hydrolysis of organic compounds with phosphoester bonds at phosphate limitation.

• Klíčová slova
• Acid phosphatase, Glucose-6-phosphate, Overproducing strain, Pho5, Polyphosphate, Saccharomyces cerevisiae,
• MeSH
• kyselá fosfatasa genetika   izolace a purifikace   metabolismus   MeSH
• polyfosfáty metabolismus   MeSH
• promotorové oblasti (genetika) genetika   MeSH
• rekombinantní fúzní proteiny genetika   izolace a purifikace   metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny genetika   izolace a purifikace   metabolismus   MeSH
• Saccharomyces cerevisiae enzymologie   metabolismus   MeSH
• substrátová specifita MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kyselá fosfatasa MeSH
• PHO5 protein, S cerevisiae MeSH Prohlížeč
• polyfosfáty MeSH
• rekombinantní fúzní proteiny MeSH
• Saccharomyces cerevisiae - proteiny MeSH