Enzyme Activity Measurement for Alanine Dehydrogenase

Creative Enzymes has been researching and developing methods for the catalytic activity quantification of enzymes for many years, and we have enough confidence to offer the enzyme activity assay for alanine dehydrogenase. Our passion for utmost quality and perfect service is only exceeded by our marvelous reputation in the marketplace.

Alanine dehydrogenase (EC 1.4.1.1) is an enzyme that catalyzes the reversible conversation of pyruvate and ammonia to L-alanine, while simultaneously generating NADH, which acts as a hydrogen donor for many biosynthetic processes. The reductive amination of pyruvate is significant in that it leads to alanine biosynthesis and provides a means of incorporating free ammonia into cellular protein. The systematic name of this enzyme class is L-alanine:NAD⁺ oxidoreductase (deaminating), which is also known as:

• AlaDH;
• L-alanine dehydrogenase;
• NAD⁺-linked alanine dehydrogenase;
• alpha-alanine dehydrogenase;
• NAD⁺-dependent alanine dehydrogenase;
• alanine oxidoreductase;
• NADH-dependent alanine dehydrogenase

Alanine dehydrogenase activity was established in a wide range of bacterial species involved in the use of resting cell suspensions and cell-free extracts. In mycobacteria, it was first identified as an enzyme absent from the vaccine strains of M. bovis BCG but present in virulent Mycobacterium tuberculosis。 It was suggested that impairment of M. bovis BCG replication in humans, due to the lack of functional alanine dehydrogenase, inhibited the development of protective immunity. Unlike alanine dehydrogenase from other bacteria, the alanine dehydrogenase from M. tuberculosis is a multispecific enzyme using either glyoxylate or pyruvate as a substrate. It catalyzes the reduction of pyruvate to alanine, or glyoxylate to glycine, coupled with the oxidation of NADH to NAD⁺. In the reverse direction, it oxidizes alanine to pyruvate (ALD), but does not use glycine as a substrate. Hence it has been proposed that alanine dehydrogenase plays an important role in maintaining redox balance during shift down of M. tuberculosis to the NRP state. Note that M. tuberculosis is the cause of tuberculosis, which is responsible for about 2 million deaths every year with as many patients as almost one-third of the world’s population.

To this point, it can be seen that alanine dehydrogenase is a considerably useful enzyme in the medical and pharmaceutical industry. Furthermore, alanine dehydrogenase may perform multiple roles, depending on the conditions or perhaps even the various species. Thus the catalytic activity quantification of this enzyme should receive more and more attention. Creative Enzymes can offer the service of catalytic activity measurement for alanine dehydrogenase by using spectrophotometric assays, which can be performed at 25°C by following the absorbance at 340 nm. By constantly striving for service of utmost quality, Creative Enzymes has earned the trust of countless customers, and is proud to offer a wide range of highly sophisticated research services. Creative Enzymes is your best choice for any research or development activities involving alanine dehydrogenase.

Figure: The crystal structure of apo-L-alanine dehydrogenase from M. Tuberculosis.
PDB: 2VHY