Capsid catalysis : de novo enzymes on viral proteins
Thesis: Ph. D., Massachusetts Institute of Technology, Department of Biological Engineering, 2015.
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| Format: | Thesis |
| Language: | eng |
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Massachusetts Institute of Technology
2015
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| Online Access: | http://hdl.handle.net/1721.1/99052 |
| _version_ | 1826192155659468800 |
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| author | Casey, John P., Jr |
| author2 | Angela M. Belcher. |
| author_facet | Angela M. Belcher. Casey, John P., Jr |
| author_sort | Casey, John P., Jr |
| collection | MIT |
| description | Thesis: Ph. D., Massachusetts Institute of Technology, Department of Biological Engineering, 2015. |
| first_indexed | 2024-09-23T09:07:05Z |
| format | Thesis |
| id | mit-1721.1/99052 |
| institution | Massachusetts Institute of Technology |
| language | eng |
| last_indexed | 2024-09-23T09:07:05Z |
| publishDate | 2015 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/990522019-04-10T20:50:13Z Capsid catalysis : de novo enzymes on viral proteins Casey, John P., Jr Angela M. Belcher. Massachusetts Institute of Technology. Department of Biological Engineering. Massachusetts Institute of Technology. Department of Biological Engineering. Biological Engineering. Thesis: Ph. D., Massachusetts Institute of Technology, Department of Biological Engineering, 2015. Cataloged from PDF version of thesis. Includes bibliographical references (pages 107-119). Biocatalysis has grown rapidly in recent decades as a solution to the evolving demands of industrial chemical processes. Mounting environmental pressures and shifting supply chains underscore the need for novel chemical activities, while rapid biotechnological progress has greatly increased the utility of enzymatic methods. Enzymes, though capable of high catalytic efficiency and remarkable reaction selectivity, still suffer from relative instability, high costs of scaling, and functional inflexibility. Herein, M13 bacteriophage libraries are engineered as a biochemical platform for de novo semisynthetic enzymes, functionally modular and widely stable. Carbonic anhydrase-inspired hydrolytic activity via Zn²+ coördination is first demonstrated. The phage clone identified hydrolyzes a range of carboxylic esters, is active from 25°C to 80°C, and displays greater catalytic efficacy in DMSO than in water. Reduction-oxidation activity is subsequently developed via heme and copper cofactors. Heme-phage complexes oxidize multiple peroxidase substrates in a pH-dependent manner. The same phage clone also binds copper(II) and oxidizes a catechol derivative, di-tert-butylcatechol, using atmospheric oxygen as a terminal oxidant. This clone could be purified from control phage via Cu-NTA columns, enabling future library selections for phage that coördinate Cu²+ ions. The M13 semisynthetic enzyme platform complements biocatalysts with characteristics of heterogeneous catalysis, yielding high-surface area, thermostable biochemical structures readily adaptable to reactions in myriad solvents. As the viral structure ensures semisynthetic enzymes remain linked to the genetic sequences responsible for catalysis, future work could tailor the biocatalysts to high-demand synthetic processes by evolving new activities, utilizing high-throughput screening technology and harnessing M13's multifunctionality. by John P. Casey, Jr. Ph. D. 2015-09-29T19:00:23Z 2015-09-29T19:00:23Z 2015 2015 Thesis http://hdl.handle.net/1721.1/99052 921844972 eng MIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission. http://dspace.mit.edu/handle/1721.1/7582 119 pages application/pdf Massachusetts Institute of Technology |
| spellingShingle | Biological Engineering. Casey, John P., Jr Capsid catalysis : de novo enzymes on viral proteins |
| title | Capsid catalysis : de novo enzymes on viral proteins |
| title_full | Capsid catalysis : de novo enzymes on viral proteins |
| title_fullStr | Capsid catalysis : de novo enzymes on viral proteins |
| title_full_unstemmed | Capsid catalysis : de novo enzymes on viral proteins |
| title_short | Capsid catalysis : de novo enzymes on viral proteins |
| title_sort | capsid catalysis de novo enzymes on viral proteins |
| topic | Biological Engineering. |
| url | http://hdl.handle.net/1721.1/99052 |
| work_keys_str_mv | AT caseyjohnpjr capsidcatalysisdenovoenzymesonviralproteins |