Presentation Title

Characterization of Poly(Aspartic Acid) Hydrolase-2

Location

College of Science and Mathematics (COSM)

Session Format

Poster Presentation

Co-Presenters, Co- Authors, Co-Researchers, Mentors, or Faculty Advisors

Dr. Mitch Weiland, Faculty Advisor

Abstract

Poly(carboxylates) are water-soluble synthetic materials that function as superabsorbent materials, commonly found in diapers and feminine hygiene products. Unfortunately, these materials are non-biodegradable and there is a need to find alternatives to mitigate environmental accumulation. Poly(aspartic acid) is a biodegradable alternative to poly(carboxylates), and has been shown to be broken down by three different enzymes. This project focuses on one of these poly(aspartic acid) degrading proteins, PAAH-2, and aims to characterize the catalytic active site of the hydrolytic enzyme. The DNA coding for the protein PAAH-2 was purchased and expressed in E.coli cells, which are used to produce the protein. The protein was purified using an immobilized metal affinity column and the purity was assessed by SDS-PAGE gels, following an established protocol. Initial crystallization leads were identified by the Hauptmann-Woodward Institute (HWI) and PAAH-2 crystallization experiments were optimized using the purified protein and varying the buffer conditions around those initially seen by HWI. Here we will present the structure of PAAH-2 along with activity assays used to characterize this protein.

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Creative Commons License
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Presentation (Open Access)

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Characterization of Poly(Aspartic Acid) Hydrolase-2

College of Science and Mathematics (COSM)

Poly(carboxylates) are water-soluble synthetic materials that function as superabsorbent materials, commonly found in diapers and feminine hygiene products. Unfortunately, these materials are non-biodegradable and there is a need to find alternatives to mitigate environmental accumulation. Poly(aspartic acid) is a biodegradable alternative to poly(carboxylates), and has been shown to be broken down by three different enzymes. This project focuses on one of these poly(aspartic acid) degrading proteins, PAAH-2, and aims to characterize the catalytic active site of the hydrolytic enzyme. The DNA coding for the protein PAAH-2 was purchased and expressed in E.coli cells, which are used to produce the protein. The protein was purified using an immobilized metal affinity column and the purity was assessed by SDS-PAGE gels, following an established protocol. Initial crystallization leads were identified by the Hauptmann-Woodward Institute (HWI) and PAAH-2 crystallization experiments were optimized using the purified protein and varying the buffer conditions around those initially seen by HWI. Here we will present the structure of PAAH-2 along with activity assays used to characterize this protein.