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Enhancing Protein Activity and Stability via Scaffold Engineerin

Posted on:2019-05-24Degree:Ph.DType:Dissertation
University:Rensselaer Polytechnic InstituteCandidate:Basore, Danielle AnnaFull Text:PDF
GTID:1440390002993240Subject:Biochemistry
Abstract/Summary:
The discipline of protein engineering has incredible potential to change the world, in the medical and environmental fields, as well as others. This powerful tool can be used to design novel vaccine antigens to prevent diseases and other medical conditions and can also be used to redesign enzymes to improve them for use in green chemistry initiatives.;Rational and computational design methods have been applied to the problem of creating an immunocontraceptive against sperm. Based on the CatSper complex, a voltage gated calcium channel found only in sperm, this contraceptive will reversibly block fertilization long-term, by inducing an immune response that will cause any sperm present to be targeted and destroyed. The CatSper peptides are carried on the surface of a virus like particle composed of the major capsid protein of the human papilloma virus, L1.;In a similar vein, a chimeric VLP vaccine antigen has been designed to provide universal protection from the dengue virus. 350 million people are infected with this virus every year, and while a vaccine from Sanofi-Pasteur has just been approved, it does not offer complete and universal protection against all serotypes of the virus. We have proposed a construct composed of the fusion loop of the envelope protein of dengue virus, a highly conserved region of the protein known to stimulate a neutralizing antibody response.;The recycling of plastics is an ever-present problem that is only growing worse over time. Current methods involve harsh chemicals, high temperatures, and long times. Protein engineering can provide a solution in the form of specially modified enzymes, designed to break down plastics such as PET, commonly found in soda bottles. The enzyme cutinase is a powerful hydrolase which naturally works on the substrate cutin, a waxy coating found on fruit and leaves. It can also work on PET, but the high temperatures required to render the PET amorphous inactivate the enzyme. With both rational and computational techniques, we redesigned this enzyme to be more thermostable.
Keywords/Search Tags:Protein, PET
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