Structural and Functional Characterization of PAA Hydrolase PAAH-2
Location
Presentation- College of Science and Mathematics
Document Type and Release Option
Thesis Presentation (Archived)
Faculty Mentor
Dr. Mitch Weiland
Faculty Mentor Email
mweiland@georgiasouthern.edu
Presentation Year
2021
Start Date
26-4-2021 12:00 AM
End Date
30-4-2021 12:00 AM
Keywords
Georgia Southern University, Honors Symposium, Presentation
Description
Poly(carboxylate), a water-soluble polymer, has many applications, one of which is as a superabsorbent material commonly found in products like diapers and feminine hygiene products. These materials are non-biodegradable and alternatives are necessary to mitigate their accumulation in the environment. Poly(aspartic acid) (PAA) is a safe, biodegradable alternative that can be broken down by three different enzymes. This project focuses on PAAH-2, one of the PAA degrading enzymes, and aims to characterize the catalytic active site of the hydrolase. The DNA sequence for the protein was purchased and then expressed using E. coli cells. The protein was purified using an immobilized metal affinity column and the purity was assessed using SDS-PAGE. Initial crystallization leads were identified by the Hauptmann-Woodward Institute (HWI) and PAAH-2 crystallization experiments were optimized using the conditions initially seen by HWI. Here we will present the structure of PAAH-2 predicted from the crystal structure as well as the activity assays used to characterize the protein.
Academic Unit
College of Science and Mathematics
Structural and Functional Characterization of PAA Hydrolase PAAH-2
Presentation- College of Science and Mathematics
Poly(carboxylate), a water-soluble polymer, has many applications, one of which is as a superabsorbent material commonly found in products like diapers and feminine hygiene products. These materials are non-biodegradable and alternatives are necessary to mitigate their accumulation in the environment. Poly(aspartic acid) (PAA) is a safe, biodegradable alternative that can be broken down by three different enzymes. This project focuses on PAAH-2, one of the PAA degrading enzymes, and aims to characterize the catalytic active site of the hydrolase. The DNA sequence for the protein was purchased and then expressed using E. coli cells. The protein was purified using an immobilized metal affinity column and the purity was assessed using SDS-PAGE. Initial crystallization leads were identified by the Hauptmann-Woodward Institute (HWI) and PAAH-2 crystallization experiments were optimized using the conditions initially seen by HWI. Here we will present the structure of PAAH-2 predicted from the crystal structure as well as the activity assays used to characterize the protein.
Comments
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