The Immobilization of Lipases on Porous Support by Adsorption and Hydrophobic Interaction Method

Abstract

<jats:p>Four major enzymes commonly used in the market are lipases, proteases, amylases, and cellulases. For instance, in both academic and industrial levels, microbial lipases have been well studied for industrial and biotechnological applications compared to others. Immobilization is done to minimize the cost. The improvement of enzyme properties enables the reusability of enzymes and facilitates enzymes used in a continuous process. Immobilized enzymes are enzymes physically confined in a particularly defined region with retention to their catalytic activities. Immobilized enzymes can be used repeatedly compared to free enzymes, which are unable to catalyze reactions continuously in the system. Immobilization also provides a higher pH value and thermal stability for enzymes toward synthesis. The main parameter influencing the immobilization is the support used to immobilize the enzyme. The support should have a large surface area, high rigidity, suitable shape and particle size, reusability, and resistance to microbial attachment, which will enhance the stability of the enzyme. The diffusion of the substrate in the carrier is more favorable on hydrophobic supports instead of hydrophilic supports. The methods used for enzyme immobilization also play a crucial role in immobilization performance. The combination of immobilization methods will increase the binding force between enzymes and the support, thus reducing the leakage of the enzymes from the support. The adsorption of lipase on a hydrophobic support causes the interfacial activation of lipase during immobilization. The adsorption method also causes less or no change in enzyme conformation, especially on the active site of the enzyme. Thus, this method is the most used in the immobilization process for industrial applications.</jats:p>

Biocatalysis for Rare Ginsenoside Rh2 Production in High Level with Co-Immobilized UDP-Glycosyltransferase Bs-YjiC Mutant and Sucrose Synthase AtSuSy

“…For the co-immobilization of engineered M315F and AtSuSy onto heterofunctional resin with two kinds of groups, the two-step immobilization protocol was accomplished according to Mateo et al [32,33]. His tag affinity method is generic and can be easily applied to specific adsorption [28,34]. The crude two enzymes from E. coli BL21(DE3) crude cell extract (M315/AtSuSy total activity addition of 4000 mU/g wet resin) were in 10 mL solution (25 mM Tris-HCl buffer, pH 7.0) at 25 • C and adsorbed by heterofunctional resin (1.0 g wet resin) under mild stirring for 6 h. In the first step, the optimal ratio of M315F/AtSuSy adsorbed by resin for the biosynthesis of Rh2 was determined using the various ratios of M315F/AtSuSy adjustment, next, adsorption time, pH, and temperature were studied, respectively.…”

Modulation of the Biocatalytic Properties of a Novel Lipase from Psychrophilic Serratia sp. (USBA-GBX-513) by Different Immobilization Strategies

“…Lipases are the most used enzymes in biocatalysis due to their high activity, specificity, selectivity, and robustness in a variety of reaction media [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 ]. Lipases have a peculiar catalytic mechanism, called interfacial activation, which is based on the interaction of the lipase with hydrophobic structures, which causes deep conformational changes on the structure of the active site of the lipase and its consequent enzyme activation [ 25 , 26 , 27 , 28 , 29 ], permitting the enzyme to act in the hydrolysis of insoluble drops of oils and be adsorbed on any hydrophobic surface [ 30 , 31 , 32 , 33 , 34 ].…”

“…A significant number of reversible and irreversible immobilization protocols have been developed to immobilize lipases [ 12 , 31 , 34 , 35 , 36 , 37 , 38 , 39 ]. Such immobilization strategies can improve enzyme stability and even the enantioselectivity or enantiospecificity of the biocatalyst if properly performed [ 16 ].…”

“…Immobilization methods included: interfacial adsorption on octyl agarose hydrophobic support (OC) [ 17 ], covalent binding on octyl glyoxyl Sepharose (OCGLX) after interfacial activation immobilization [ 44 , 45 , 46 ], direct covalent binding on CNBr-Sepharose [ 62 , 63 ], and anion exchange on Q-Sepharose [ 61 ]. The immobilized enzyme derivatives were compared with the commercial lipase B from Candida antarctica (CALB), immobilized on octyl Sepharose [ 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , ...…”

Stabilization of β-Galactosidase on Modified Gold Nanoparticles: A Preliminary Biochemical Study to Obtain Lactose-Free Dairy Products for Lactose-Intolerant Individuals

“…Recently, nanoparticles have also attracted the attention of researchers to improve traditional enzyme immobilization. Nanoparticle-based enzyme immobilization is an enigma that deserves special attention, as it provides a greater surface area for binding higher amounts of enzyme to the matrix, and prevents unfolding of protein and permits greater flexibility for the conformational changes required for enzyme activity [ 17 , 18 ]. The other advantages involved with its use include continuous operations, catalyst recycling, enhanced stability, easy separation from the reaction mixture, possible modulation of the catalytic properties, and much easier prevention of microbial growth [ 19 , 20 ].…”