PPA E.Coli

The recombinant form of inorganic pyrophosphatase is typically derived from the bacterium Escherichia coli (E. coli). This enzyme is a homohexameric protein, meaning it consists of six identical subunits, each containing 175 amino acid residues . The recombinant version is often engineered to include a His-tag, which facilitates its purification and enhances its stability .

Inorganic pyrophosphatase is essential for numerous biosynthetic pathways, including the synthesis of nucleic acids (DNA and RNA), proteins, and polysaccharides. During these processes, pyrophosphate is released as a byproduct, and its accumulation can inhibit further reactions. By hydrolyzing pyrophosphate into orthophosphate, inorganic pyrophosphatase helps to maintain the thermodynamic balance and ensures the continuation of these vital biosynthetic reactions .

Recombinant inorganic pyrophosphatase from E. coli is widely used in molecular biology and biochemistry research. It is particularly valuable in in vitro transcription reactions to increase RNA yield and enhance DNA replication . The enzyme’s activity is dependent on the presence of magnesium ions (Mg²⁺), which act as cofactors .

The recombinant enzyme is relatively thermostable and can be stored as a lyophilized powder at -20°C for extended periods. Upon reconstitution, it remains stable at -20°C . The enzyme is supplied in a Tris-buffered solution containing protease inhibitors to prevent degradation .

Recombinant inorganic pyrophosphatase is rigorously tested to ensure the absence of contaminants such as endonucleases, exonucleases, RNases, and alkaline phosphatase. This high level of purity is crucial for its use in sensitive biochemical assays and molecular biology applications .