Biochemical aspects of lipase immobilization at polysaccharides for biotechnology

Abstract

The design of immobilized enzyme preparations is an important and relevant area of modern sciences and technologies. Immobilization of enzymes from animal sources (component I) on natural carriers (component II) increases the system stability by protecting the active site of the enzyme from deactivation; facilitates the separation and accelerates the recovery of the enzyme. This makes reuse possible and provides a significant reduction in operating costs. Hydrolytic enzymes (such as lipases) and polysaccharides (such as chitosan) are the most promising of such pairs of components. The main attention here is devoted to the discussion on lipase immobilization on polysaccharide (mainly - chitin and chitosan). Based on the analysis of the available literature, the most adequate method is the immobilization of lipase from porcine pancreas (LPP) on polysaccharide particles (such as chitin or chitosan) pre-treated with ultrasound (to increase the particle surface area) and glutaraldehyde (for particle activation) that shows reasonably high LPP activity and stability. In order to increase further the activity of the lipase, some authors proposed to incorporate a spacer in the form of 1,3-diaminopropane (or 1,3-diaminobutane) prior to activation of the surface of the chitosan particles. In particular cases, the use of chitin (instead of chitosan) may be an alternative solution for biotechnological applications. Recently the idea of constructing "supramolecular enzyme systems" realized in the so-called "coimmobilized multienzymatic systems" strategy. The most fascinating example is the combined assay of a mixture of native LPP, glycerol kinase (from Cellulomonas) and glycerol-3-phosphate oxidase (from Aerococcus viridans) linked by glutaraldehyde to chitosan (as shell for inorganic nanoparticle core). This material was placed on a Pt-electrode as biosensor and was successfully applied for amperometric determination of the triglyceride level in the serum of healthy and diseased person. Thus, the whole innovative research-production sequence is described by Aggarwal V. and Pundir C.S.: from simple components to advanced material and further biomedical application. Thus, the following approach of lipase immobilization appears the most promising for future applications: a few types of lipases or the combination of LPP with some other enzymes immobilized simultaneously on multifunctional carriers (as nanohybrids of inorganic core and polysaccharide shell).

Keywords: Chitin and chitosan; Colloid biochemistry; Enzyme preparations; Lipase immobilization; Nanoparticles.