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The acid phosphatase Pho5 of Saccharomyces cerevisiae is not involved in polyphosphate breakdown

  • Nadeshda Andreeva
  • Larisa Ledova
  • Lubov Ryasanova
  • Tatiana KulakovskayaEmail author
  • Michail Eldarov
Original Article
  • 48 Downloads

Abstract

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.

Keywords

Acid phosphatase Pho5 Polyphosphate Saccharomyces cerevisiae Overproducing strain Glucose-6-phosphate 

Notes

Acknowledgements

The authors thank Elena Makeeva for her help with preparing the manuscript.

Funding

This study was supported in part by the Russian Foundation for Basic Research (grant no. 17-04-00822).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2019

Authors and Affiliations

  1. 1.FRC Pushchino Center for Biological Research, Skryabin Institute of Biochemistry and Physiology of MicroorganismsRussian Academy of SciencesMoscow regionRussia
  2. 2.Institute of BioengineeringResearch Center of Biotechnology of the Russian Academy of SciencesMoscowRussia

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