Lipase supports

Mixtures of triglycerides, triglycerides plus free fatty adds or triglycerides plus fatty acid alkyl esters are used as reactants in fat modification processes. These mixtures are exposed to lipases supported on macroporous particles in the presence of a small amount of water. Liquid substrates (oils) can be reacted without use of a solvent, but with solid reactants (fats) it is necessary to add a solvent to ensure that the reactants and products are completely dissolved in the organic phase. Various water immisdble solvents can be used, but hexane is preferred for commercial operation because this solvent is already used industrially for the processing of oils and fats. 

In this communication a study of the catalytic behavior of the immobilized Rhizomucor miehei lipase in the transesterification reaction to biodiesel production has been reported. The main drawbacks associated to the current biodiesel production by basic homogeneous catalysis could be overcome by using immobilized lipases. Immobilization by adsorption and entrapment have been used as methods to prepare the heterogeneous biocatalyst. Zeolites and related materials have been used as inorganic lipase supports. To promote the enzyme adsorption, the surface of the supports have been functionalized by synthesis procedures or by post-treatments. While, the enzyme entrapping procedure has been carried out by sol-gel method in order to obtain the biocatalyst protected by a mesoporous matrix and to reduce its leaching after several catalytic uses. 

Theil, F. (1995) Lipase-supported synthesis of biologically-active compounds. Chem. Reviews., 95, 2203-2227. 

Bosley, J.A. and Clayton, J.C. (1993) Blueprint for a lipase support Use of hydrophobic controlled-pore glasses as model systems. Biotechnology and Bioengineering, 43, 934-938. 

Fig. 7.5 Pseudomonas cepacia lipase supported on ceramic particles PS-C catalysed polyester synthesis using diethyl octane-1,8-dicarboxylate and 1,4-butanediol in [bmim]PF6 at room...

Bosley, J. A., and J. C. Cla)4on. 1994. Blneprint for a Lipase Support Use of Hydrophobic Controlled-Pore Glasses as Model Systems. Biotechnology and Bioengineering 43 (10) 934-938. 

Camilo Naranjo, J., Cordoba, A., Giraldo, L., Garcia, V.S., Carlos Moreno-Pirajan, J., 2010. Lipase supported on granular activated carbon and activated carbon cloth as a catalyst in the synthesis of biodiesel fuel. Journal of Molecular Catalysis B-Enzymatic 66, 166-171. 

Anionic - surfactants, such as - fatty alcohol sulfates, fatty alcohol ether sulfates and ->sulfo-succinates, are also applied. Proteolytic - enzymes and sometimes lipases support the process. Dehairing and Liming ... 

In addition to having the required spedfidty, lipases employed as catalysts for modification of triglycerides must be stable and active under the reaction conditions used. Lipases are usually attached to supports (ie they are immobilised). Catalyst activity and stability depend, therefore, not only on the lipase, but also the support used for its immobilisation. Interesterification reactions are generally run at temperatures up to 70°C with low water availability. Fortunately many immobilised lipases are active and resistant to heat inactivation under conditions of low water availability, but they can be susceptible to inactivation by minor components in oils and fats. If possible, lipases resistant to this type of poisoning should be selected for commercial operations. 

Lipases catalyse reactions at interfaces, and to obtain a high rate of interesterification the reaction systems should have a large area of interface between the water immiscible reactant phase and the more hydrophilic phase which contains the lipase. This can be achieved by supporting the lipase on the surface of macroporous particles. 

Adsorption on solid matrices, which improves (at optimal protein/support ratios) enzyme dispersion, reduces diffusion limitations and favors substrate access to individual enzyme molecules. Immobilized lipases with excellent activity and stability were obtained by entrapping the enzymes in hydrophobic sol-gel materials [20]. Finally, in order to minimize substrate diffusion limitations and maximize enzyme dispersion, various approaches have been attempted to solubilize the biocatalysts in organic solvents. The most widespread method is the one based on the covalent linking of the amphiphilic polymer polyethylene glycol (PEG) to enzyme molecules [21]. 

Immobilization of lipase on porous ceramic support (Toyonite) for acceleration ... 

Lipase from C. rugosa entrapped on polyethylene glycol Iso-octane-water (4/1) Olive oil hydrolysis 35 41.5 /xmol/(min g of support) 0.043% w/v 121... 

Two possible pathways for the biosynthesis of 2-AG have been proposed (1) a phospholipase C (PLC) hydrolysis of membrane phospholipids followed by a second hydrolysis of the resulting 1,2-diacylglycerol by diacylglycerol lipase or (2) a phospholipase Ai (PLA,) activity that generates a lysophospholipid, which in turn is hydrolyzed to 2-AG by lysophospholipase C (Fig. 5) (Piomelli, 1998). Alternative pathways may also exist from either triacylglycerols by a neutral lipase activity or lysophosphatidic acid by a dephosphorylase. The fact that PLC and diacylglycerol lipase inhibitors inhibit 2-AG formation in cortical neurons supports the contention that 2-AG is, at least predominantly, biosynthesized by the PLC pathway (Stella, 1997). However, a mixed pathway may also be plausible. 

Blanco, R.M., Terreros, P., Fernandez-Perez, M., Otero, C. and az-Gonzalez, G. (2004) Functionalization of mesoporous silica for lipase immobilization Characterization of the support and the catalysts. Journal of Molecular Catalysis B-Enzymatic, 30, 83-93. 

One of the most promising applications of enzyme-immobilized mesoporous materials is as microscopic reactors. Galameau et al. investigated the effect of mesoporous silica structures and their surface natures on the activity of immobilized lipases [199]. Too hydrophilic (pure silica) or too hydrophobic (butyl-grafted silica) supports are not appropriate for the development of high activity for lipases. An adequate hydrophobic/hydrophilic balance of the support, such as a supported-micelle, provides the best route to enhance lipase activity. They also encapsulated the lipases in sponge mesoporous silicates, a new procedure based on the addition of a mixture of lecithin and amines to a sol-gel synthesis to provide pore-size control. 

Layered phosphate/phosphonate and phosphonate materials, obtained by substitution of the phosphate moiety by phosphonate groups, display interesting tunable hydrophilic/organophilic properties for adsorption processes. When Candida rugosa lipase (CRL) is simply equilibrated with zirconium phosphate and phosphonate [135,136], immobilization was demonstrated to take place at the surface of the microcrystals. However, because lipase exhibits a strong hydrophobic character, its uptake by zirconium phosphate and phosphonate was much more related to the hydrophobic/hydrophilic character of the supports than to the surface area properties. A higher uptake is observed for zirconium-phenylphosphonate (78 %)... 

The concept of zeolite action was tested in a particular reaction where the enzyme is exposed from the beginning to an acidic environment the esterification of geraniol with acetic acid catalyzed by Candida antarctica lipase B immobilized on zeolite NaA [219]. Lipases have been used for the hydrolysis of triglycerides and due to their ambivalent hydrophobic/hydrophilic properties they are effective biocatalysts for the hydrolysis of hydrophobic substrates [220]. When water-soluble lipases are used in organic media they have to be immobilized on solid supports in order to exhibit significant catalytic activity. 

In conventional synthetic transformations, enzymes are normally used in aqueous or organic solvent at moderate temperatures to preserve the activity of enzymes. Consequently, some of these reactions require longer reaction times. In view of the newer developments wherein enzymes can be immobilized on solid supports [183], they are amenable to relatively higher temperature reaction with adequate pH control. The application of MW irradiation has been explored with two enzyme systems namely Pseudomonas lipase dispersed in Hyflo Super Cell and commercially available SP 435 Novozym (Candida antarctica lipase grafted on an acrylic resin). 

The multipolymer enzymatic resolution of soluble polymer-supported alcohols 42 and 43 was achieved using an immobilised lipase from Candida Antarctica (Novozym 435). The R-alcohol was obtained in enantiomerically pure form (>99% ee) after its cleavage from the poly(ethylene) glycol (PEG) scaffold . The achiral hydantoin- and isoxazoline-substituted dispirocyclobutanoids 47 were produced using both solution and solid-phase synthesis <00JOC3520, OOCC1835>. 

Lipases can catalyze hydrolysis of esters, synthesis of esters, trans-esterification, and synthesis of some polymers. They have been applied in many fields including the food industry, fine chemistry, and the pharmaceutical industry. The low stability of native lipases makes them unsuitable for industrial applications. In order to use them more economically and efficiently, their operational stability can be improved by immobilization. Numerous efforts have been focused on the preparation of lipases in immobilized forms involving a variety of both support materials and immobilization methods [278],... 

J. Xu et al. [283] have shown that immobilization of enzymes can be done using a specially designed composite membrane with a porous hydrophobic layer and a hydrophilic ultrafiltration layer. A polytetrafluoroethylene (PTFE) membrane with micrometer pores as an excellent hydrophobic support for immobilization was employed for the porous hydrophobic layer, and a biocompatible material of polyvinyl alcohol (PVA) which provided a favourable environment to retain the lipase activity was used to prepare the hydrophilic... 

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