PPA Yeast

Inorganic pyrophosphatase from yeast is typically produced using recombinant DNA technology. The enzyme is encoded by the ppa gene from Saccharomyces cerevisiae (yeast) and is expressed in Escherichia coli (E. coli) for large-scale production . The recombinant enzyme is then purified to achieve high purity levels, often exceeding 95% as determined by SDS-PAGE .

The enzyme is a homodimer, consisting of two identical subunits with a molecular weight ranging between 32-35 kDa . It operates optimally at a pH of around 7.2 and requires magnesium ions (Mg²⁺) as a cofactor for its catalytic activity . The reaction it catalyzes can be represented as:

$$\text{PPi} + \text{H}_2\text{O} \rightarrow 2 \text{Pi}$$

Inorganic pyrophosphatase plays a vital role in cellular metabolism by preventing the accumulation of inorganic pyrophosphate, which can be inhibitory to various biochemical pathways. Some key applications include:

• RNA and DNA Synthesis: The enzyme enhances the yield of RNA in transcription reactions by hydrolyzing PPi, a byproduct of nucleotide incorporation .
• Nucleic Acid Amplification: It is used in nucleic acid amplification experiments to promote the reaction equilibrium towards product formation, thereby increasing the amount of the desired product .
• Metabolic Pathways: It is involved in various metabolic pathways where PPi is generated as a byproduct, ensuring these reactions proceed efficiently .

The enzyme is typically stored at -20°C in a buffer containing 20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, and 50% glycerol . This storage condition ensures the enzyme remains stable and retains its activity over time.