Grantee Research Project Results
Final Report: Development of aromatic dehalogenases as a tool to remediate halogenated aromatic pollutants
EPA Grant Number: SU836121Title: Development of aromatic dehalogenases as a tool to remediate halogenated aromatic pollutants
Investigators: Kharel, Madan Kumar
Institution: University of Maryland - Eastern Shore
EPA Project Officer: Page, Angela
Phase: I
Project Period: September 1, 2015 through August 31, 2016
Project Amount: $15,000
RFA: P3 Awards: A National Student Design Competition for Sustainability Focusing on People, Prosperity and the Planet (2015) RFA Text | Recipients Lists
Research Category: P3 Awards , Pollution Prevention/Sustainable Development , Sustainable and Healthy Communities , P3 Challenge Area - Chemical Safety
Objective:
The main objective of our proposed project is to develop an environmentally friendly system to expedite the biodegradation of hard-to-degrade environmental pollutants, particularly halogenated aromatic compounds (HACs). Agrochemicals such as herbicides (Atrazine) and pesticides (Chlorpyrifos) have been routinely used in the farms to maintain good agricultural productivity. However, many of the chemicals are resistant to degradation under the normal environmental conditions. The routine release of such chemicals and their persistence in the environment have been posing a serious risk both to public health and to the environment. This project is dedicated to developing a dehalogenase enzyme system to facilitate the degradation of these agrochemicals without causing additional negative environmental impact. The project utilizes a stepwise enzyme evolution approach to improve catalytic efficiency of select native bacterial dehalogenases. The medium throughput screening system proposed in this project will be useful in selecting robust mutants generated through directed evolution process. We propose that production of such enzyme in soil-dwelling bacterial species will expedite the degradation of structurally diverse HACs under a natural environment.
Summary/Accomplishments (Outputs/Outcomes):
Gene sequence of hydrolytic dehalogenase that is capable of removing halogen atom from aromatic rings was synthesized. The gene was cloned in pET-28a vector to produce a 6×Histidine-tagged recombinant protein. Conditions for overexpression of protein in Escherichia coli have been optimized. We recently purified the recombinant protein using immobilized metal affinity chromatography (IMAC). Currently, we are testing the activity of the purified enzyme and are also setting up 96-well plate-based medium throughput screening system for dehalogenation activity.
Journal Articles:
No journal articles submitted with this report: View all 1 publications for this projectSupplemental Keywords:
Sustainable bioremediation, halogenated pollutants, P3 challenge area-built environment, P3 challenge area-materials and chemicalsThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.