Enzyme Activity Measurement for Fructose-Bisphosphatase

Creative Enzymes is an industrial biotech company specialized in enzyme activity assays. True to our company’s tradition, we firmly believe that the highest-quality standard for our work and services is the key to our global success and high-level customer satisfaction. Herein, we are proud to offer the highly sophisticated enzymatic assay for fructose-bisphosphatase.

Fructose-bisphosphatase (EC 3.1.3.11; D-fructose-1,6-bisphosphate 1-phosphohydrolase) is an enzyme that catalyzes the rate-limiting step in the gluconeogenic pathway, namely the hydrolysis of fructose-1,6-bisphosphate to fructose-6-phosphate and inorganic phosphate. Fructose-bisphosphatase is regarded as one of the key enzymes in gluconeogenesis. Fructose-2,6-bisphosphate and AMP inhibit fructose-bisphosphatase synergistically and can potentially limit the entire gluconeogenic pathway by their effect on fructose-bisphosphatase. Fructose-bisphosphatase requires divalent cations (Mg²⁺, Mn²⁺, or Zn²⁺) for catalysis, and some monovalent cations such as K⁺ or NH_{4+ can further enhance activity. This enzyme has been characterized from bacteria, yeast, as well as higher eukaryotes. The amino acid sequences of mammalian enzymes are 85% identical to each other, and are similar to the Class I fructose-bisphosphatase in bacteria. In Escherichia coli, two fructose-bisphosphatases have been identified encoded by the fbp gene and glpX gene that belong to the Class I and Class II, respectively. A very divergent Class III fructose-bisphosphatase has also been identified in Bacillus subtilis. Interestingly, fructose-bisphosphatase activity has also been detected in cell extracts of several archaea, though orthologous genes with structural similarity to previously reported fructose-bisphosphatase are not present on their genomes. A new group of fructose-bisphosphatases (class V) is found in thermophilic archaea and the hyperthermophilic bacterium Aquifex aeolicus.}

Note that, porcine fructose-bisphosphatase, which is the most extensively studied of all fructose-bisphosphatases, is a homotetramer with subunit molecular mass of 37 kDa. Functionally, fructose-bisphosphatase can serve as a target for drug development in the treatment of type 2 diabetes mellitus. Besides, fructose-bisphosphatase also plays a pivotal role in biotechnology, molecular biology, and biosynthesis. Thus, these significant functions promote a strong demand for monitoring the activity of fructose-bisphosphatase.

Creative Enzymes is capable of making the most reliable enzymatic assay available for fructose-bisphosphatase. The enzyme activity is measured spectrophotometrically by following the reduction of NADP⁺ in the presence of glucose-6-phosphate isomerase (EC 5.3.1.9) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49). The formation of fructose 6-phosphate results in the reduction of an equivalent amount of NADP⁺, which is monitored by measuring the increase in optical density at 340 nm. Our high-quality enzyme assay services are most trusted and have been commended by countless customers. As your trustworthy partner, Creative Enzymes will be continue to sustain the professional standard and provide efficient services in the future.

Figure: The crystal structure of porcine fructose-bisphosphatase.
PDB: 1NUY