Anuseema Bhadauriya

Project

Kinetic versus thermodynamic stability: Bacillus subtilis lipase A as a test case

Info

Area: biocatalysis
Project Start: 1.07.2013
Supervisor(s): Prof. Dr. Holger Gohlke
University: Duesseldorf

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Aims of the project

This project aims at understanding factors that influence the kinetic stability of a protein and making use of this knowledge for improving kinetic stability in the context of rational protein engineering. Bacillus subtilis lipase A (BSLA) will be used as a test system, for which thermodynamically and kinetically stabilizing mutants have been characterized and for which a complete saturation library has been constructed (group of Prof. K.-E. Jaeger). Selected mutants of these datasets will be analyzed by means of computational structural bioinformatics/biophysics approaches with respect to thermostability, local unfolding events, exposure of hydrophobic patches and cleavage sites for proteases. These analyses will be complemented by differential scanning calorimetry and proteolytic degradation experiments (collaboration with the group of Prof. K.E. Jaeger), NMR spectroscopy and X-ray crystallography (collaboration with the group of Prof. D. Willbold). From these results a set of rules shall be derived with which new BSLA mutants with improved kinetic stability can bepredicted. These mutants shall be constructed and experimentally tested.

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Anuseema’s project
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