GOR E.Coli

Glutathione oxidoreductase (GOR), also known as glutathione reductase (GR), is an essential enzyme in maintaining the redox balance within cells. It belongs to the class-I pyridine nucleotide disulfide oxidoreductase family. The recombinant form of this enzyme, produced in Escherichia coli (E. coli), is widely used in research and industrial applications due to its high purity and activity.

GOR is a single, non-glycosylated polypeptide chain consisting of 473 amino acids, with a molecular mass of approximately 51.2 kDa . The enzyme is fused to a 23 amino acid His-tag at the N-terminus, which facilitates its purification through chromatographic techniques . The active site of GOR contains a redox-active disulfide bond, which is crucial for its catalytic function.

The primary role of GOR is to maintain high levels of reduced glutathione (GSH) in the cytosol. It achieves this by catalyzing the reduction of oxidized glutathione (GSSG) to GSH, using NADPH as an electron donor . This reaction is vital for protecting cells against oxidative stress and maintaining the redox balance within the cell.

GOR operates through a well-defined mechanism involving the transfer of electrons from NADPH to GSSG. The enzyme’s active site disulfide bond undergoes a series of redox reactions, ultimately leading to the reduction of GSSG to GSH. This process is essential for the detoxification of reactive oxygen species (ROS) and the maintenance of cellular homeostasis .

The recombinant form of GOR is produced in E. coli using advanced genetic engineering techniques. The enzyme is expressed as a soluble protein and purified to a high degree of purity (>90%) using affinity chromatography . The purified enzyme is typically formulated in a buffer containing Tris-HCl, glycerol, NaCl, and dithiothreitol (DTT) to ensure its stability and activity .

GOR is stable when stored at 4°C for short-term use (2-4 weeks) and at -20°C for long-term storage. To prevent degradation, it is recommended to add a carrier protein such as human serum albumin (HSA) or bovine serum albumin (BSA) and avoid multiple freeze-thaw cycles .

Recombinant GOR is widely used in biochemical and biomedical research to study redox biology, oxidative stress, and related cellular processes. It is also employed in industrial applications, including the production of reduced glutathione for pharmaceutical and cosmetic purposes .