Lipophilization Based On Enzyme Retention

As anthocyanin lipophilization emerged as an efficient technique to improve their enzyme chemical stability; liposolubility and antioxidant properties for novel technological applications. Protein extract, rich in lipase B, in powder form from Candida antarctica was kindly donating by Genofocus.

Enzyme retention in the membrane is due to enzyme adsorption at the pore walls of the membrane support; and to enzyme retention in the bulk of the porous matrix; whose leakage from the membrane is minimized by the denser active layer at the opposite membrane side.

Lipophilization method

The enzyme retention was conducting by permeation of the CalB-rich extract at constant transmembrane pressures; (TMP) of 0.04 bar and 0.4 bar by cell pressurization with an inert gas (N2) stream; in order to evaluate the influence of TMP on membrane
enzyme loading.

Washing of the membranes used in the esterification reaction was further performing upon enzyme retention for an easier removal of unbound matter. The influence of membrane washing in the reaction performance was accessing comparing a reaction performing with and without a membrane washing procedure. Study showing the stability of the protein retention and robustness of the enzymatic membrane system.

Enzymatic esterification

However, the membrane P10 with retained enzyme was installing; in a cross-flow cell and washed by recirculation of the buffer solution. The enzymatic esterification of Cy3glc with octanoic acid was studied using the membrane and transmembrane pressure, which led to the highest protein loading, i.e. using membrane enzymatic systems prepared by filtration of CalB-rich extract through a P10 membrane, at a TMP of 0.04 bar.

But the study reporting for the first time; immobilization studies of a CalB-rich extract in different membrane systems and describes an efficient and alternative method for the synthesis of lipophilic anthocyanins. The results showed that it was possible to increase the enzymatic activity of the CalB-rich extract without the need of further costly and exhaustive processing steps for enzyme purification, just by simple retention of the crude extract in an adequate asymmetric ultrafiltration membrane.