E.coli Pyrophosphatase (50 U)

A recombinant enzyme from QIAGEN that catalyzes the Mg²⁺-dependent hydrolysis of pyrophosphate (P₂O₇⁴⁻ + H₂O → 2 HPO₄²⁻), enhancing the efficiency of nucleotide synthesis. It is widely used in in vitro transcription and RNA synthesis reactions to reduce inhibitory effects of pyrophosphate (PPi), with a starting recommendation of 0.1–1 units per mL to identify the optimal concentration.

The protein is expressed in a recombinant E. coli strain carrying the E. coli pyrophosphatase gene and has a molecular weight of 19.7 kDa. One unit is defined as the amount of enzyme that liberates 1 µmol of phosphate per minute from inorganic pyrophosphate at 37°C and pH 8.5. Quality specifications include SDS-PAGE purity of >95%, specific activity of 3,500 U/mg, single-stranded exonuclease <1.0% released, double-stranded exonuclease <1.0% released, double-stranded endonuclease showing no conversion, and E. coli DNA contamination of <10 copies. Activity is Mg²⁺-dependent, making Mg²⁺ an essential component of the reaction buffer.

The enzyme is supplied at 100 U/mL in 20 mM Tris-HCl, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol (pH 8.0 at 25°C) and stored at -25°C to -15°C. It is intended for in vitro use only.