In this study we characterized and cloned the PSHAa1385 gene in Escherichia coli. We also characterized the recombinant protein by biochemical and biophysical methodologies.
The PSHAa1385 gene sequence showed a significant homology with several carboxyl-esterase and acetyl-esterase genes from γ-proteobacteria genera and yeast. The recombinant protein exhibited a significant activity towards pNP-acetate, α-and β-naphthyl acetate as generic substrates, and 4-methylumbelliferyl p-trimethylammonio cinnamate chloride (MUTMAC) as a specific substrate, indicating that the protein exhibits a feruloyl esterase activity that it is displayed by similar enzymes present in other organisms.
Finally, a three-dimensional model of the protein was built and the amino acid residues involved in the catalytic function of the protein were identified.