Thermostable Mutation of Halohydrin Dehalogenase in Proteins and Possible Types of Variance

Omer Uzen
Omer Uzen

Some parts of the proteins were reacted with various chemicals, microorganisms and some substance derivatives. He examined the mutation and variants of this mutation that occurred as a result of these reactions. As a result of his research, he reacted to these unexplored events as a result of his experimental, simulation and sampling thesis studies with different combinations of some protein regions.

Ömer UZEN, a researcher and scientist from Turkey, contributed to the world of science with this study and discovery.

The main goal of the Thermostable Mutant of Halohydrin Dehalogenase is to create a robust variant of halohydrin dehalogenase that can be used in this way. In this research, a computational workflow was used to improve the stability of halohydrin dehalogenase from Agrobacterium Radiobacter, a homotetrameric enzyme of the short chain dehydrogenase reductase superfamily with 27 kilodalton (kDa) subunits. It was used to engineer halohydrin dehalogenase mutants to improve stability in water-organic solvent mixtures. It is possible to combine functional mutations into a single variant to study point mutations, remove non-functional ones, confirm selected mutations, and generate the desired enzyme. Using 1P×O, H-he-C crystal structure, possible point mutations and changes in folding free energy were investigated with FoldX.

Omer Uzen

Halohydrin dehalogenase is an enzyme that catalyzes the halogenation of halohydrins to produce the corresponding epoxides. In this study, it was aimed to increase the catalytic ability of halohydrin dehalogenase in the presence of organic co-solvents. In this context, stabilizing mutations were investigated. The high activity and stability of enzymes are very advantageous for laboratory applications. Enzymes with good stability can be used as a good catalyst at elevated temperatures and in the presence of organic co-solvents. 1P×O: Human cyclin-dependent kinase 2 inhibitor compared to [4-(2 Amino-4-methyl-thiazol-5-yl)-pyrimidin-2-yl]-(3-nitro-phenyl)-amine. Cyclin-dependent kinase, or CDK, is a type of enzymatic protein found in eukaryotic cells and plays a key role in a number of biological processes in cellular metabolism and regeneration, collectively referred to as the cell cycle. Many protein kinases are also classified as genes and make up about 2 percent of all human genes. The mechanism of cyclin-dependent kinase activity is based on the phosphorylation process or the addition of phosphate groups to the protein surface. However, for the protein to be modified by phosphorylation, the protein must form a complex form of the type known as a cyclin. This is why this specialized protein is called a cyclin-dependent kinase.

FoldX provides a rapid and quantitative estimation of the importance of the interactions that contribute to the stability of proteins and protein complexes. The predictive power of FoldX was tested on a very large point mutant (1088 mutants) covering most of the structural environments found in proteins. FoldX uses a complete atomic description of the structure of proteins. The different energy terms considered in FoldX have been weighted using empirical data from protein engineering experiments. The current energy function uses minimal computational resources and can therefore be easily used in protein design algorithms and in the field of protein structure and folding pathways prediction where a fast and accurate energy function is required.

Various simulations were performed for each mutant examined to generate beneficial mutations. Next, genetic engineering techniques were used for protein expression and purification. Dehalogenase activities and halide release were measured by various methods. Activity, heat resistance and co-solvent tolerance were measured using different methods. There were two mutants named H-he-C-C153N and H-he-C-H12. In this structure, we observed a mutant named C153N with 5Kwe current electrical waves placed above the 153 code. It is a mutant effective in increasing the apparent opening temperature.

Omer Uzen

When the simulations were analyzed according to the results of this scientific study, a total of 23 point mutations were found to be stabilized in 29 different regions. The most effective mutation was C153N. This mutant causes a +13 ⁰C increase in the apparent opening temperature. When the crystal structure of the C153N mutant was examined, it was observed that there was a chloride ion in its active site. In the structure of H-he-C-C153N, 4 monomers were found in asymmetrical form. The thermal stability provided by the C153N mutant is due to the hydrogen bonding network containing regular solvent molecules.

📩 28/08/2022 10:12

Be the first to comment

Leave a response

Your email address will not be published.