Related Experiment Videos
A kinetic epoxidation assay for chloroperoxidase
1Department of Biochemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, USA.
Biochemical and Biophysical Research Communications
|December 22, 1999
Summary
Chloroperoxidase (CPO) catalyzes enantioselective epoxidation. A new spectrophotometric assay rapidly measures CPO activity by monitoring indene disappearance, aiding mutant screening.
Area of Science:
- Biocatalysis
- Enzyme kinetics
- Spectrophotometry
Background:
- Chloroperoxidase (CPO) is known for diverse enantioselective epoxidation reactions.
- Current methods for evaluating CPO activity, primarily gas chromatography, are complex and time-consuming.
Purpose of the Study:
- To develop a rapid and convenient spectrophotometric assay for measuring chloroperoxidase (CPO) activity.
- To provide a high-throughput method for assessing enzyme function in mutant screening.
Main Methods:
- Spectrophotometric monitoring of indene disappearance at 250 nm.
- Utilizing the rate of indene consumption as an index of CPO catalytic activity during epoxidation.
Main Results:
- A straightforward spectrophotometric assay for CPO activity was successfully established.
- The assay allows for rapid and convenient evaluation of enzyme performance.
Conclusions:
- The developed spectrophotometric assay offers a significant improvement over traditional methods for CPO activity assessment.
- This assay is highly valuable for large-scale screening of chloroperoxidase mutants.