AbstractStreptomyces fradiaevar. k11 is a Gram-positive soil microorganism capable of degrading chicken feathers. Apart from being mostly protein, chicken feathers have a considerable level of lipids, with wax esters being the largest lipid class. The waxes may pose a challenge while rendering the feathers into coproducts, such as feather meal, and so the identification of a wax-ester hydrolase is warranted. A draft genome sequence ofS. fradiaevar. k11 was used to identify 14 gene sequences of potential lipid-degrading enzymes. The genes were expressed inE. coliBL21(DE3) cells on a pET vector and screened for activity. Four of the 14 enzymes had detectable activity, with two of the enzymes, SFK3309 and SFK3087, active against p-nitrophenyl palmitate, a representative water-insoluble substrate. A modified enzymatic assay was designed to measure activity against three model wax substrates: jojoba oil, beeswax, and cetyl-palmitate. SFK3309 was characterized to hydrolyze all three wax substrates. Kinetic experiments for SFK3309 were performed with cetyl-palmitate at 37°C, pH 8.0. TheKmwas determined to be 850 µM and theKcatwas 11.63 s-1. Through the characterization of SFK3309 as a wax-ester hydrolase, biotechnological implications of wax ester hydrolases in the rendering of many industrial wastes can be substantiated for further studies.
Preparation of cetyl palmitate-based PEGylated solid lipid nanoparticles by microfluidic technique
