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
• storage condition: OK to freeze
• concentration: ≥100 units/mL
• color: colorless
• optimum pH: 9.0 (25 °C)
• pH: 8.0 (25 °C)
• solubility: soluble; water: miscible
• suitability: suitable for molecular biology
• UniProt accession no.: P0A7A9
• application(s): research use
• foreign activity: DNAse, none detected; RNAse, none detected; Nickase, none detected
• shipped in: dry ice
• storage temp.: -10 to -25°C

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).

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. 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.

Biochem/physiol Actions

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

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