Enzyme Activity Measurement of Protocatechuate 3,4-Dioxygenase Using Spectrophotometric Assays

At Creative Enzymes, providing exactly approaches for oxidoreductases activity measurement has never been a challenge, thanks to our specialized experts with rich experiences and advanced equipment with precise outputs. The assay service for protocatechuate 3,4-dioxygenase is performed via spectrophotometric analysis, giving reliable results.

Protocatechuate 3,4-dioxygenase (EC 1.13.11.3) belongs to the intradiol dioxygenases (IDOs) class, which uses the Fe³⁺ cofactor to achieve ortho cleavage between the hydroxyl functions. This enzyme acts on 3,4-dihydroxybenzoate (PCA) or related catecholic compounds, catalyzeing the cleavage of O₂ and the concomitant ring opening to yield cis, cis-muconate products. It has been isolated from many divergent soil bacteria and employed as a reagent to improve organic fluorophore-stability.

This enzyme participates in benzoate degradation via hydroxylation and 2,4-dichlorobenzoate degradation, which are very important part in aerobic biodegradative pathways that allow the organisms to derive all their carbon and energy from aromatic hydrocarbons. After the cleavage by protocatechuate 3,4-dioxygenase, aromatic cyclic structures are opened and the linear-structured hydrocarbons are produced. The product enters the citrate cycle after a series of catalysis steps. By this group of enzymes, complicated chemical compounds are broken down to simple molecules for easy utilization in the center metabolism to generate energy, which is vital for the full use of any substance to sustain organisms’ living. Protocatechuate 3,4-dioxygenase also contributes to environment protection. Owning to the broad spectrum of enzyme types, these enzymes can be used in bioremediation of polluted groundwater or soil contaminated with various aromatic compounds. Because of the promising ability for pollution depuration, purification and substrate specificity of the enzyme are targeted by many studies. On one hand, purified enzymes for high catalysis activity accelerate the re-clean cycle; on the other hand, a wide range of substrate specificity allows applications in more changeable environments. Creative Enzymes offers proper assays focused on this enzyme using most reliable spectrophotometric analysis.

Figure: The structure of protocatechuate 3,4-dioxygenase complexed with 3-hydroxybenzoate.
Reference: Orville, A.M.et al. Biochemistry 1997 36: 10039-10051.

Creative Enzymes is devoted to providing the best enzyme activity assays. We have the most advanced equipment and professional teams, which give the accurate test results. A majority of customers are highly content with the high-quality and rapid services. Creative Enzymes will continue the excellent support to your research in the future.