Inhibition of Peroxidase Activity in Horseradish
Faculty Sponsor
Dr. Deanna Ojennus
Session Type
Poster Presentation
Research Project Abstract
The purpose of this study was to analyze inhibitors of horseradish peroxidase activity for the development of an undergraduate enzyme kinetics laboratory. Horseradish is commonly used in techniques such as ELISA due to its monomeric nature and the ease at which it produces colored products. Horseradish peroxide (HRP) was extracted in a phosphate-citrate buffer for this study and activity monitored through UV-VIS. Peroxidase activity was measured using a Tetratmethylbenzidine substrate (TMB, 0.1 mg/mL + 0.0006% H2O2) and a 96 well-plate visible light spectrometer. Cobalt (II) chloride (CoCl2, 0.25 mM) and Sodium azide (NaN3, 0.02%) were tested, as two inhibitors of HRP, to determine what type of inhibition would be taking place. Peroxidase inhibitors CoCl2 and NaN3 displayed non-competitive inhibition while competitive and uncompetitive inhibition was not observed. Adoption of 96-well plate readers appear to be promising for enzyme kinetic studies in the biochemistry labs as a teaching tool.
Session Number
PS2
Location
HUB Multipurpose Room
Abstract Number
PS2-k
Inhibition of Peroxidase Activity in Horseradish
HUB Multipurpose Room
The purpose of this study was to analyze inhibitors of horseradish peroxidase activity for the development of an undergraduate enzyme kinetics laboratory. Horseradish is commonly used in techniques such as ELISA due to its monomeric nature and the ease at which it produces colored products. Horseradish peroxide (HRP) was extracted in a phosphate-citrate buffer for this study and activity monitored through UV-VIS. Peroxidase activity was measured using a Tetratmethylbenzidine substrate (TMB, 0.1 mg/mL + 0.0006% H2O2) and a 96 well-plate visible light spectrometer. Cobalt (II) chloride (CoCl2, 0.25 mM) and Sodium azide (NaN3, 0.02%) were tested, as two inhibitors of HRP, to determine what type of inhibition would be taking place. Peroxidase inhibitors CoCl2 and NaN3 displayed non-competitive inhibition while competitive and uncompetitive inhibition was not observed. Adoption of 96-well plate readers appear to be promising for enzyme kinetic studies in the biochemistry labs as a teaching tool.
