SAE0215

Inorganic Pyrophosphatase from Escherichia coli

≥100 units/mL, buffered aqueous solution

• UNSPSC Code: 12352204

Properties

• biological source: Escherichia coli
• Quality Level: 200
• recombinant: expressed in E. coli
• grade: for molecular biology
• description: Recombinant, expressed in E.coli
• assay: ≥95% (size exclusion chromatography)
• form: buffered aqueous solution
• specific activity: ≥100 units/mL
• shelf life: 2 yr at -20 °C ((retest))
• mol wt: 19.7 kDa

Description

General description

Inorganic pyrophosphatase (PPase) is a ubiquitous enzyme that catalyzes pyrophosphate hydrolysis. It plays an important role in energy metabolism by providing a thermodynamic pull for biosynthetic reactions, such as protein, RNA, and DNA synthesis. Nucleic acid synthesis would be energetically impossible in vivo if not coupled with the hydrolysis of pyrophosphate (PPi)[1][2].

Application

This product is based on the native pyrophosphatase from E. coli, Uniprot No. P0A7A9. Pyrophosphatase in E. coli is a homohexameric protein containing 175 amino acids residues per subunit[3]. This product is a recombinant protein expressed in E. coli and induced by IPTG. Each subunit has a MW of 19.7 kDa and theoretical pI of ~5. The protein activity is Mg2+ dependent and it is a relatively thermostable protein[4].

Biochem/physiol Actions

Inorganic pyrophosphatase (PPase) is a ubiquitous enzyme that catalyzes pyrophosphate to phosphate.[5] It plays an important role in energy metabolism as it provides a thermodynamic pull for biosynthetic reactions, such as protein, RNA, and DNA synthesis.[6]

Features and Benefits

This product has a purity minimum of 95% (SEC-HPLC) and an activity minimum of 100 units/mL to enhance RNA yield during transcription.

Unit Definition

One unit will release 1.0 µmole of inorganic orthophosphate per minute at pH 9 at 25 °C. The reaction buffer used for determination of enzyme activity contains 50 mM Tris-HCl, pH 9.0.

Physical form

The product is supplied as an aqueous solution containing 20mM Tris-HCl, 100mM NaCl, 1mM DTT, 0.1mM EDTA, and 50% glycerol, titrated to pH 8 at 25 °C.

Safety Information

• Storage Class: 10 - Combustible liquids
• wgk_germany: WGK 1