Critical Role of Different Immobilized Biocatalysts of a Given Lipase in the Selective Ethanolysis of Sardine Oil

Abstract

Different immobilized derivatives of two lipases were tested as catalysts of the synthesis of ethyl esters of omega-3 fatty acids during the ethanolysis of sardine oil in solvent-free systems at 25 °C. Lipases from Thermomyces lanuginosus (TLL) and Lecitase Ultra (a phospholipase with lipolytic activity) were studied. Lipases were adsorbed on hydrophobic Sepabeads C18 through the open active center and on an anion-exchanger Duolite with the active center exposed to the reaction medium. TLL-Sepabeads derivatives exhibit a high activity of 9 UI/mg of immobilized enzyme, and they are 20-fold more active than TLL-Duolite derivatives and almost 1000-fold more active than Lipozyme TL IM (the commercial derivative from Novozymes). Lecitase-Sepabeads exhibit a high selectivity for the synthesis of the ethyl ester of EPA that is 43-fold faster than the synthesis of the ethyl ester of DHA.

Modulation of the regioselectivity of Thermomyces lanuginosus lipase via biocatalyst engineering for the Ethanolysis of oil in fully anhydrous medium

“…On the other hand, the least active biocatalyst preserves the 1,3-selectivity of TLL and they are obtained with DVB resins. It seems that the adsorbed open structures of TLL may be slightly different [ 27 , 36 ]. They are not depending on the resin (eg., Purolite) but they strongly depend on the hydrophobic moieties (C18 versus DVB groups).…”

“…TLL is a 1,3-regioselective and fairly stable lipase with excellent properties to catalyze transesterification processes [ 1 , 14 ]. TLL adsorbed on hydrophobic supports is a very interesting immobilized lipase biocatalyst to perform transesterification of oils [ 25 – 27 ]. In this case, TLL is adsorbed through the open active center fixed on the hydrophobic surface of the hydrophobic supports (Fig.…”

Immobilization of Lipase from Penicillium sp. Section Gracilenta (CBMAI 1583) on Different Hydrophobic Supports: Modulation of Functional Properties

“…Immobilization of lipases occurs via a peculiar mechanism known as interfacial activation [ 7 , 8 ], This naturally-occurring mechanism was used as a tool to immobilize several microbial lipases in many different hydrophobic supports proving to be a simple and efficient method that can allow selective purification, due to lipase adsorption even at low ionic strength, giving rise to purified-stabilized derivatives [ 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 ].…”

“…Toyopearl, an acrylic support formed by thin crossed fibers that might be even smaller than the lipase molecules, also possesses certain hydrophobicity. Lewatit and Sepabeads are macroporous cross-linked methacrylate polymers, which might allocate lipases inside their structure [ 12 , 16 ]. Moreover, coating the fiber surfaces with butyl, phenyl or octyl groups may reinforce the hydrophobic character of each matrix.…”

“…This open form is quite unstable in aqueous and homogenous medium, but becomes stabilized by adsorption on hydrophobic surfaces, which can be originated from a substrate or hydrophobic supports. Adsorption on hydrophobic supports has been widely used for lipase immobilization, since the technique is rapid, simple, and of low cost and high selectivity, allowing at one time purification, immobilization, hyperactivation, and stabilization [ 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 ]. A limitation of this type of biocatalyst, however, is the enzyme release from the support which can occur in the presence of some reaction products, an important factor that should be taken in consideration during the selection of an industrial lipase biocatalyst [ 17 ].…”