The use of protocatechuate dioxygenase for maintaining anaerobic conditions in biochemical experiments.
The study of redox-active systems often requires the maintenance of anaerobic conditions. The glucose oxidase system has often been used to maintain anaerobic conditions, but it has some drawbacks, such as the production of H(2)O(2) and limitations on stability. Protocatechuate dioxygenase from Burkholderia cepacia and the substrate, protocatechuate, constitute an alternate effective oxygen-scrubbing system that can be used in a wide variety of biochemical experiments. We have shown its suitability for maintaining rigorous anaerobic environments in solutions of pH 6-9, at temperatures from 4 to 35 degrees C, and for periods of time up to 15 months. The enzyme system was shown to be stable under these conditions and effective for maintaining anaerobic conditions in titrations of FAD. It is also suitable for scrubbing various types of apparatus such as stopped-flow instruments for anaerobic experiments.
Patil
P V
PV
Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan, 48109-0606, USA.
Ballou
D P
DP
D000693:Anaerobiosis; D016956:Burkholderia cepacia / Q000201:enzymology; D002623:Chemistry Techniques, Analytical / Q000379:methods; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D010084:Oxidation-Reduction; D011520:Protocatechuate-3,4-Dioxygenase / Q000378:metabolism*; D013696:Temperature
D011520:Protocatechuate-3,4-Dioxygenase