CaltechTHESIS
  A Caltech Library Service

Characterization of the functional diversity and evolvability of chimeric enzymes assembled by structure-guided SCHEMA recombination

Citation

Carbone, Martina Nini (2010) Characterization of the functional diversity and evolvability of chimeric enzymes assembled by structure-guided SCHEMA recombination. Dissertation (Ph.D.), California Institute of Technology. http://resolver.caltech.edu/CaltechTHESIS:05272010-170914354

Abstract

In nature proteins evolve by a combination of point mutagenesis and recombination. This process has generated hundreds of fascinating and structurally complex protein folds capable of performing a myriad of important and diverse biochemical functions. This has inspired protein engineers to mimic natural protein evolution in the library to construct synthetic proteins with new or improved properties. Here I show that homologous protein recombination can be used in the library to engineer novel enzymes with new catalytic activities and altered substrate specificities. I also propose that homologous recombination can be used in the laboratory to overcome the challenge of improving the native activities of wild-type proteins. In nature recombination may have helped proteins escape local maxima of the fitness landscape by introducing many homologous mutations to which proteins are highly tolerant. Protein engineers can possibly use it for the same purpose. I validate this hypothesis computationally with highly simplified protein models, and I attempt an experimental verification of this theory with cellulases.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:protein engineering, protein evolution, protein recombination, lattice proteins
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Chemical Engineering
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Arnold, Frances Hamilton
Thesis Committee:
  • Adami, Christoph Carl
  • Davis, Mark E.
  • Tirrell, David A.
  • Arnold, Frances Hamilton (chair)
Defense Date:18 May 2010
Funders:
Funding AgencyGrant Number
National Science FoundationUNSPECIFIED
National Institutes of HealthR01-GM068664-0
Record Number:CaltechTHESIS:05272010-170914354
Persistent URL:http://resolver.caltech.edu/CaltechTHESIS:05272010-170914354
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5867
Collection:CaltechTHESIS
Deposited By: Martina Carbone
Deposited On:06 Jun 2011 16:03
Last Modified:26 Dec 2012 03:26

Thesis Files

[img]
Preview
PDF (thesis) - Final Version
See Usage Policy.

11Mb

Repository Staff Only: item control page