Enzyme Activity Measurement of Uronate Dehydrogenase Using Spectrophotometric Assays

Creative Enzymes is a leading company in providing activity assay services for oxidoreductases. Our decade-long experiences are based on the thorough study on assay methods and the in-depth learning from a large amount of finished testing. We have a high-standard measurement control which guarantees the most reliable results for uronate dehydrogenase.

Until now, uronate dehydrogenases (EC 1.1.1.203) has only been found in Agrobacterium and Pseudomonas. This enzyme belongs to the family of oxidoreductase and catalyzes the oxidation of D-galacturonic acid to D-galactaro-1,5-lactone, or D-glucuronic acid to D-glucaro-1,5-lactone, accompanied with the conversion from NAD⁺ to NADH. Other names in common use are uronate: NAD-oxidoreductase and uronic acid dehydrogenase.

D- Galacturonic acid is the main component of pectin, which is an important raw material for biotechnology applications, especially having a potential in bio-conversions to fuels and fine chemicals. There are two different catabolic pathways involved in D-galacturonic acid catabolism; one of them is an oxidative pathway. In this pathway, D-galacturonic acid is first oxidized into meso-galactaric acid and then converted in the following step to α-ketoglutarate. Uronate dehydrogenase (EC 1.1.1.203) is the key enzyme in the oxidative pathway of D-galacturonic acid catabolism in bacteria. Up to now, only uronate dehydrogenase from Agrobacterium tumefaciens (AtUdh) has been studied in details. AtUdh belongs to the short-chain dehydrogenase/reductase (SDR) superfamily and accepts both D-galacturonic acid and D-glucuronic acid as substrates with similar affinities. Other studies showed that the catalytic activity requires Mg²⁺ ions. Further research beyond basic characterization on this enzyme is very limited. Much mechanistic and structural basics need to be understood before the enzyme can be used in bio-conversion. Proper activity assays are the initial step to experiment on uronate dehydrogenases from other species. Creative Enzymes provides spectrophotometric assay services to satisfy customers’ demands of using uronate dehydrogenase in the next generation biotechnology products.

Figure: The crystal structure of uronate dehydrogenases from Agrobacterium tumefaciens.
Reference: Tarja Parkkinen et al. J. Biol. Chem. 2011 286: 27294-27300

Creative Enzymes is one of the few companies that provide assay services for uronate dehydrogenases. We have established suitable assay methods for purified enzymes and also for enzyme extractions. Our results are secured with reliable quantification approaches and consistent reproducibility. In the future, Creative Enzymes will continue to support your research in a rapid and efficient way.