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Pyrophosphatase, Inorganic (from yeast)

LO certified Not inactivated at 80°C in 20 min Recombinant enzyme

Pyrophosphatase, Inorganic catalyzes the hydrolysis of inorganic pyrophosphate to two orthophosphates.
  
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Thermo Scientific Pyrophosphatase, Inorganic catalyzes the hydrolysis of inorganic pyrophosphate into two orthophosphates (see Figure 1 in Supporting data). The enzyme requires a divalent metal cation with Mg2+ conferring the highest activity (see Reference 1).

Highlights

Applications

  • High yield RNA synthesis by in vitro transcription (see Reference 2)
  • DNA polymerization reactions: preventing accumulation of pyrophosphate (see References 3, 4)
  • Removal of contaminant PPi in reagents used for SNP genotyping by methods based on the detection of pyrophosphate (see Reference 5)

Note

The enzyme can be diluted with the supplied storage (dilution) buffer. Use of this enzyme in certain applications may be covered by patents and may require a license.

  
Storage Condition-20 C
HazardousNo
Concentration0.1 u/µL
Definition of Activity Unit
  • One unit of the enzyme hydrolyzes 1 µmol of inorganic pyrophosphate in 1 minute at 25°C.
  • Enzyme activity is assayed in the following mixture: 100 mM Tris-HCl (pH 7.2), 2 mM MgCl2 and 2 mM inorganic pyrophosphate (PPi).
Inhibition and Inactivation
  • Inhibitors: imidodiphosphate, alpha, omega-glycol disphosphates, methanedial diphosphate, 1,2-ethanedial diphosphate (see Reference 6).
  • Inactivation by heating is not complete, reliably removed by spin column or phenol/chloroform extraction.
Molecular WeightThis enzyme is a homodimer. It consists of two identical subunits of 32 kDa.
Quality ControlThe absence of endo-, exodeoxyribonucleases, and ribonucleases confirmed by appropriate quality tests.
SourceE. coli cells with a cloned ppa gene of Saccharomyces cerevisiae.
Storage (Dilution) BufferThe enzyme is supplied in: 10 mM Tris-HCl (pH 7.5), 0.1 mM EDTA and 50% (v/v) glycerol.
Use of Pyrophosphatase in preventing inhibition of na synthesis by phyrophosphate

Use of Pyrophosphatase in preventing inhibition of na synthesis by phyrophosphate

Use of Pyrophosphatase in preventing inhibition of na synthesis by phyrophosphate

Use of Pyrophosphatase, Inorganic, in preventing inhibition of nucleic acid synthesis by phyrophosphate.
Pyrophosphatase prevents inhibition of polymerization reaction by hydrolysis of pyrophosphate accumulated during reaction.


References

  1. B. S. Cooperman, The mechanism of action of yeast inorganic pyrophosphatase. Meth. Enzymol. 87, 526-548 (1982).
  2. P. R. Cunningham et al., Use of inorganic pyrophosphatase to improve the yield of in vitro transcription reactions catalyzed by T7 RNA polymerase. Biotechniques. 9, 713-714 (1990).
  3. S. Tabor et al., DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Effect of pyrophosphorolysis and metal ions.  J. Biol. Chem. 265, 8322-8328 (1990).
  4. B. F. Dean et al., Rapid amplification of plasmid and phage DNA using phi29 DNA polymerase and multiply-primed Rolling Circle amplification. Genome Res. 11, 1095-1099 (2001).
  5. G. H. Zhou et al., Quantitative detection of single nucleotide polymorphisms for a pooled sample by a bioluminometric assay coupled with modified primer extension reactions (BAMPER). Nucleic Acids Res.29, E93 (2001).
  6. J. W. Sperow et al., Yeast Inorganic Pyrophosphatase. The Journal of Biological Chemistry. 6, 2062-2065 (1973).

Protocols and Product Manuals

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