2.1 To study the protein sequence of Coelacanth uricase (LM-UOX) using multiple sequences alignment. This tool helps to identify conserved regions/residues, which infer the structure and the identity of the protein to uricases from other organisms.
2.2 To study the protein sequence of LM-UOX using molecular phylogenetics. The results from these tools infer the evolutionary relationship between Coelacanth and other organisms
2.3 To predict the 3D structures of the wild type LM-UOX and its mutants (D287C and N288C). The results from these studies indicate structure-activity relationship and, perhaps, biochemical properties of the proteins such as solubility and thermostability.
2.4 To produce recombinant wild type LM-UOX and its mutants (D287C and N288C) in E. coli. The process includes gene cloning, optimization for protein expression and protein purification.
2.5 To characterize biochemical properties of recombinant wild type LM-UOX and its mutants (D287C and N288C) such as natural molecular weight, peptide mass finger print, enzymatic activity and kinetic behaviors, optimum pH and temperature of catalysis, and thermostability.
2.6 To study whether the mutants (D287C and N288C) contain disulfide bond at the mutated residues and prove whether it confer thermostability to the proteins.
