Enhanced catalytic stability of lipase immobilized on oxidized and disulfide-rich eggshell membrane for esters...

Eggshell membrane (ESM) is an industrial waste that is available in abundance from food industry. Present study investigated the physicochemical properties of oxidized ESM and compared the efficiency of ESM and oxidized ESM as carrier for Burkholderia cepacia lipase (BCL) used in esters hydrolysis and transesterification. Following oxidation treatment, FTIR analysis and Ellman's assay showed amino acid cysteine in ESM was oxidized to form disulfide bond-containing cystine. In addition, AFM analysis showed ESM which exhibited a highly porous filamentous structure appeared to be coalesce following oxidation treatment. Oxidized ESM also showed reduced porosity (38.67%) in comparison to native ESM (51.65%). BCL were successfully immobilized on oxidized ESM through carrier activation method (enzyme loading of 5.01mg protein/g oxidized ESM). These immobilized lipase demonstrated significantly (P<0.05) enhanced catalytic stability with close to 100% of initial hydrolysis (12.03±0.29mmol/min/g) activity; and more than 85% of its initial transesterification (7.83±0.05) activity for at least 10 consecutive runs. Enhanced catalytic stability of BCL immobilized on oxidized ESM might be due to stabilization of the protein structure in oxidized ESM by disulfide bonds which helped formation of a stable bonding with BCL.