Kinetic Characterization of 4-Hydroxyphenylacetate 1-Hydroxylase
Advisor Information
John Conrad
Location
Dr. C.C. and Mabel L. Criss Library
Presentation Type
Poster
Start Date
6-3-2015 9:00 AM
End Date
6-3-2015 10:30 AM
Abstract
Recent and rapid growth in the chemical industry has resulted in larger volumes of waste contaminating soils. Bioremediation is the use bacterial metabolic pathways to break down toxic waste into less toxic carbon compounds. The biodegradation of aromatic compounds, also a prevalent industrial waste byproduct, requires hydroxylation of the stable aromatic ring before it can be opened and further degraded. The purpose of this research was to study the activity of an under characterized enzyme that catalyzes this first step of aromatic degradation, 4- hydroxyphenylacetate 1-hydroxylase (4HPA-1H). To study the enzyme it had to first be expressed and isolated. Various methods were used to express and purify 4HPA-1H. We describe here the expression and purification of 4HPA-1H, quantified by both electrophoresis and activity assays.
Kinetic Characterization of 4-Hydroxyphenylacetate 1-Hydroxylase
Dr. C.C. and Mabel L. Criss Library
Recent and rapid growth in the chemical industry has resulted in larger volumes of waste contaminating soils. Bioremediation is the use bacterial metabolic pathways to break down toxic waste into less toxic carbon compounds. The biodegradation of aromatic compounds, also a prevalent industrial waste byproduct, requires hydroxylation of the stable aromatic ring before it can be opened and further degraded. The purpose of this research was to study the activity of an under characterized enzyme that catalyzes this first step of aromatic degradation, 4- hydroxyphenylacetate 1-hydroxylase (4HPA-1H). To study the enzyme it had to first be expressed and isolated. Various methods were used to express and purify 4HPA-1H. We describe here the expression and purification of 4HPA-1H, quantified by both electrophoresis and activity assays.