Immobilization of a Commercial Lipase from Penicillium camembertii (Lipase G) by Different Strategies

Abstract

The objective of this work was to select the most suitable procedure to immobilize lipase from Penicillium camembertii (Lipase G). Different techniques and supports were evaluated, including physical adsorption on hydrophobic supports octyl-agarose, poly(hydroxybutyrate) and Amberlite resin XAD-4; ionic adsorption on the anionic exchange resin MANAE-agarose and covalent attachment on glyoxyl-agarose, MANAE-agarose cross-linked with glutaraldehyde, MANAE-agarose-glutaraldehyde, and epoxy-silica-polyvinyl alcohol composite. Among the tested protocols, the highest hydrolytic activity (128.2 ± 8.10 IU·g−1 of support) was achieved when the lipase was immobilized on epoxy-SiO2-PVA using hexane as coupling medium. Lipase immobilized by ionic adsorption on MANAE-agarose also gave satisfactory result, attaining 55.6 ± 2.60 IU·g−1 of support. In this procedure, the maximum loading of immobilized enzyme was 9.3 mg·g−1 of gel, and the highest activity (68.8 ± 2.70 IU·g−1 of support) was obtained when 20 mg of protein·g−1 was offered. Immobilization carried out in aqueous medium by physical adsorption on hydrophobic supports and covalent attachment on MANAE-agarose-glutaraldehyde and glyoxyl-agarose was shown to be unfeasible for Lipase G. Thermal stability tests revealed that the immobilized derivative on epoxy-SiO2-PVA composite using hexane as coupling medium had a slight higher thermal stability than the free lipase.

Enzyme immobilization by adsorption: a review

“…An interesting enzyme carrier is the biodegradable and thermo-shrinkable hydroxybutyrate. It is used in the form of poly(hydroxybutyrate) nanoparticles (Deepak et al 2009) and small or large poly(hydroxybutyrate) beads (Mendes et al 2011). Another carrier, poly(o-toluidine) built of particles of o-toluidine isomer has active -NH 2 groups (Ashly and Mohanan 2010).…”

“…Lipases catalyse the hydrolysis of esters formed by short-and long-chain alcohols, mono-and multi-hydroxides, and saturated and unsaturated carboxylic acids of short and long chains. The catalysts based on lipases are used in the reactions of esterification or transesterification of different substrates (Brem et al 2011;Liu et al 2013;Yu and Fang 2013), and in the process of biodiesel production (Salis et al 2010;Mendes et al 2011;Tran et al 2012). A wide use of this group of proteins and their affinity to many carriers permit their immobilization on many organic and inorganic carriers.…”

“…Organic carriers seem to be preferred for the immobilization of lipases. They include a wide and highly diverse gamut of polymers, such as cross-linked PVA (Ozturk and Kilinc 2010) and epoxy activated PVA (Mendes et al, 2012), poly(N-methylol acrylamide) (Santos et al 2007), small and large poly(hydroxybutyrate) beads (Mendes et al 2011) and polyacrylonitryle electrospun fibres (Sakai et al 2010), organic matrices of natural origin, including chitosan beads (Nasratun et al 2010), MANAE-agarose and cellulose ultrathin film (Kosaka et al 2007), commercial polymer products, e.g. polypropylene (c) Fig.…”

Collagen-Immobilized Lipases Show Good Activity and Reusability for Butyl Butyrate Synthesis

“…Therefore, a suitable support plays a key role in the immobilization process. A variety of natural polymers have been used for lipase immobilization, e.g., agarose, chitosan, alginate, cellulose, and starch [32][33][34][35]. However, extra steps such as addition of active groups are required in these methods in order to facilitate cross-linking of these carriers with enzyme molecules.…”

Immobilization of Lipases on Magnetic Collagen Fibers and Its Applications for Short-Chain Ester Synthesis

“…Lipases have been immobilized by numerous methods such as covalent bonding, cross-linking, entrapment, encapsulation, and adsorption [9,10]. Different natural polymers have been employed for lipase immobilization such as agarose, alginate, chitosan, cellulose, and starch [11,12].…”