In Saccharomyces cerevisiae, inorganic polyphosphate is analyzed by polyphosphate extraction and subsequent quantification. Recently, we developed a method for polyphosphate quantification, and length determination of short chain polyphosphate. However, the lack of a simple, optimized and validated method for analytical polyphosphate extraction has both hindered the advance in this research field, and prevented comparability of results between laboratories. Hence, the goal of this study was to develop an analytical method for polyphosphate extraction from S. cerevisiae. Several literature methods were compared with special attention to omission of polyphosphate precipitation steps, because these work neither at low polyphosphate concentrations nor quantitatively. The best literature protocol, which takes 5.5 h and requires five reaction tubes per sample, was optimized here in regards to the amount of extracted polyphosphate and simplification of the work flow. The final protocol extracts 40 % more polyphosphate than the best literature method, takes only 30 min, requires just one reaction tube per sample, and is, therefore, proposed as the new gold standard for analytical polyphosphate extraction from S. cerevisiae. In combination with our recently published polyphosphate quantification method, total polyphosphate in S. cerevisiae can now be analyzed within 2 h.

• PMID: 30287204
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Journal Article | Research Support, Non-U.S. Gov't

Authors

Christ JJ, Blank LM

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