Serine hydrolases are one of the largest groups of enzymes and the most commonly used for biocatalysts. They are identified by the presence of a highly nucleophilic serine residue within the catalytic triad or dyad that hydrolyze substrates (Kazlauskas). BS2 from bacteria Bacillus subtilis is one known serine hydrolase that has an unselective active site, utilizing a Ser-His-Glu triad, that is able to bind with a multitude of substrates. It is believed that a mutation within the active site could decrease specificity and an increase in enzyme activity.
In this experiment, mutations L362A and F363A were studied due to their proximity to the catalytic triad within the active site (Spiller). Due to their position, it is assumed that these residues aid in enzyme efficiency and binding to the active site. By mutating the Leucine (L) and Phenylalanine (F) to Alanine (A) there should be an impact on both the substrate specificity and enzymatic efficiency. Both mutants and the wildtype were purified, isolated, and analyzed via enzyme kinetics with two ester substrates and collated to observe any differences in efficiency and specificity.
Rebuttal & Critique
There are no R & C’s yet…
Have a Rebuttal or Critique for This Work?
Ryan is an undergraduate at Butler University studying biology and biochemistry. He is from Twinsburg, Ohio Ryan is also a staff editor for research and politics at The University Journal.