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Drevon, Geraldine F (2002) ENZYME IMMOBILIZATION INTO POLYMERS AND COATINGS. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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In this study, we have developed strategies to immobilize enzymes into various polymer and coatings. Three categories of bioplastic matrices were investigated. The first type of bioplastics was prepared by irreversibly incorporating di-isopropylfluorophosphatase (DFPase) into polyurethane (PU) foams. The resulting bioplastic retained up to 67 % of the activity for native enzyme. The thermostability of DFPase was highly affected by the immobilization process. Unlike native enzyme, immobilized DFPase had biphasic deactivation kinetics. Our data demonstrated that the initial rapid deactivation of immobilized DFPase lead to the formation of a hyper-stable and still active form of enzyme. Spectroscopic studies enabled a structural analysis of the hyper-stable intermediate. Biopolymers were also prepared via atom transfer radical polymerization (ATRP) using acrylic and sulfonate-derived monomers. ATRP ensured the covalent and multi-point immobilization of enzyme within polymer matrices. However, this approach was only partially successful, as no activity retention was obtained after polymerizationEnzyme-containing PU- and Michael adduct (MA)-based coatings correspond to the last category of bioplastics that was investigated. DFPase was irreversibly incorporated into PU coatings. The distribution of immobilized DFPase as well as activity retention were homogeneous within the coating. The resulting enzyme-containing coating (ECC) film hydrolyzed DFP in buffered media at high rates retaining approximately 39% intrinsic activity. DFPase-ECC had a biphasic deactivation profile similar to that of bioplastic foams. The synthesis of enzyme-containing MA coatings was performed in a two-step process using carbonic anhydrase (CA, E.C. CA was first covalently immobilized into NVF-based water-soluble polymer (EP). The resulting EP was further entrapped into the matrix of MA coating. The so-formed ECC's exhibited approximately 7% apparent activity. CA-ECC showed good stability under ambient conditions and retained 55% activity after 90 days of storage.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Drevon, Geraldine Fgfdst1@pitt.eduGFDST1
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee Chair Russell, Alan
Committee MemberWicks, Douglas
Committee MemberBeckman, Eric
Committee MemberMatyjaszewski,
Committee MemberChapman, Toby Mtchapman@imap.pitt.eduTCHAPMAN
Committee MemberFederspiel,
Date: 2 December 2002
Date Type: Completion
Defense Date: 19 November 2002
Approval Date: 2 December 2002
Submission Date: 22 October 2002
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: Swanson School of Engineering > Chemical Engineering
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: bioplastic; coating; foam
Other ID:, etd-10222002-131628
Date Deposited: 10 Nov 2011 20:03
Last Modified: 15 Nov 2016 13:50


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