|
Mb Catalysis in Methanol
Running tests on myoglobin catalysis in methanol today. Catalysis is on the conversion of o-phenylenediamine (OPD) into 2,3-diaminophenazine (DAPN)
Spectra of the compounds in the figure below are taken from: Qi et al "Myoglobin as mimetic enzyme and its analytical application in determination of H2O2" Wuhan University Journal of Natural Sciences 2006, 11, 3, p677
Stock solutions used
- 16.4mg of o-phenylenediamine in 15.2mL of methanol
- 1.134 ml 30% H2O2 +8.866 ml Methanol
- 8.9mg of (5.7mg myoglobin +0.4861g KCl lyophilized from 20mL of 10mM citrate pH 3) in 1 mL of methanol.
Samples were made by
- 2.7 mL of the OPD solution
- 0.3 mL of the Mb solution
- Start the scan
- 1,000 us integration time
- Average over 500 scans
- Run for 10 minutes (600,000,000 us)
- Add 1mL of the H2O2 solution
- Added after scan reached 1%
- Kinetics scans were run at 5, 15, 25, 35, and 45C. Several before and after spectra were also taken, as were the dark and reference signal spectra.
- Absorbance measurements were calculated using the following equation: A = -log[(SampleSignal-DarkSignal)/(ReferenceSignal-DarkSignal)]
Shown below are kinetics traces and absorbance scans for the different temperatures
 
 
 
 
 
|
Protein Re-engineering
Main project page
Previous entry Next entry
