Enzyme Activity Measurement of Alpha, Alpha-Phosphotrehalase

As a reliable supplier, Creative Enzymes offers excellent and cost effective services, with competitive advantages over other companies. Highlights of our services include innovative test methods, fast turnovers, extensive experiences, and specialized analysis. As an industrial biotech company, Creative Enzymes can provide enzyme activity assays to support various research. Herein, we provide activity determination for alpha, alpha-phosphotrehalase.

Alpha, alpha-phosphotrehalase (EC 3.2.1.93) is encoded by treA, and is a member of the GH13 family. This enzyme catalyzes the cleavage of the alpha, alpha-1,1-glycosidic linkage of trehalose 6-phosphate to produce glucose and glucose 6-phosphate. The free glucose is further phosphorylated to the second molecule of glucose-6-phosphate, and both glucose-6-phosphate moieties later enter glycolysis, generating energy for cell growth and proliferation. This enzyme is the key part for the utilization of trehalose as a sole carbon source. Trehalose is a non-reducing disaccharide which presents in a wide variety of living organisms. With the exception of vertebrates, the sugar is distributed in nearly all other organisms including bacteria, fungi, protozoa, and plants, where it may serve as a storehouse for energy or for the synthesis of cellular components. Besides, trehalose has been shown to protect proteins and cellular membranes from inactivation or denaturation caused by desiccation, dehydration, heat, cold, and oxidation. In addition, coupled with the phosphotransferase system (PTS), trehalose is transported in cells and acts as an energy source. Up till now, only the crystal structure of alpha, alpha-phosphotrehalase from Bacillus licheniformis has been identified. Researchers are trying to obtain the structures of the enzyme from other sources to further understand the molecular basis of catalysis. Therefore, key information of the catalysis kinetics is also desired to elucidate the catalytic process, and accurate activity assays are essential to such research. Creative Enzymes performs reliable spectrophotometric assays, which has high efficiency and accuracy beyond other methods. The principle of this method is detecting the released glucose by coupling with oxidation reactions.

Figure: The crystal structure of Bacillus licheniformis trehalose-6-phosphate hydrolase (TreA), mutant R201Q, in complex with PNG.
Reference: Lin, M.-G. et al. Acta Crystallogr D Struct Biol 2016 72: 59-70.

Creative Enzymes is one of the few companies that can provide activity assay services for alpha, alpha-phosphotrehalase. The suitable assay methods are based on the comprehensive knowledge of enzyme properties. Relying on our standard progress and quality management, every test is of strict control and high reproducibility. Overall, Creative Enzymes is the best choice for contract enzyme research.