Investigation of substrate specificity of acyl-Coa thioesterase I from the bacterium Pseudomonas aeruginosa using molecular dynamics simulations

Petrović, Saša (2013) Investigation of substrate specificity of acyl-Coa thioesterase I from the bacterium Pseudomonas aeruginosa using molecular dynamics simulations. Diploma thesis, Faculty of Science > Department of Biology.

[img] PDF
Restricted to Registered users only
Language: Croatian

Download (5MB) | Request a copy

Abstract

Acyl-­CoA thioesterase I (TesA) from the bacterium Pseudomonas aeruginosa is a lipolytic enzyme that belongs to the family of GDSL hydrolases. The GDSL hydrolase family is a relatively new and not so well investigated group of enzymes with a great potential for application in pharmaceutical and food industry. Members of this enzyme family have typical GDSL motif surrounding the serine form the catalytic triad, which is different from the GXSXG motif found in other lipases. Some members of the GDSL family show large substrate promiscuity. Enzyme TesA is engaged in the fatty acid synthesis, and it hydrolyses a large number of different substrates (esters and thioesters with different fatty acid chain length). In this thesis we have tried to determine the molecular basis of substrate specificity and promiscuity of enzyme TesA using force field based computational methods, primarily molecular dynamics (MD) simulations. Simulation results are supported by experimentally determined enzyme activity towards different substrates. We have investigated the positioning of differently sized and shaped substrates in the binding site of the enzyme TesA, as well as the changes in structure and dynamics of the binding site caused by the presence of different substrates. According to the simulation results, mutations that should affect TesA activity were proposed. The proposed mutants were investigated by in silico and experimental methods.

Item Type: Thesis (Diploma thesis)
Keywords: molecular dynamics, substrate specificity, in silico mutations
Supervisor: Bertoša, Branimir
Date: 2013
Number of Pages: 46
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 29 Sep 2014 11:45
Last Modified: 29 Sep 2014 11:46
URI: http://digre.pmf.unizg.hr/id/eprint/2858

Actions (login required)

View Item View Item

Nema podataka za dohvacanje citata