Silica matrices

Porous oxide (silica) matrix infiltrated with phenolic resin... 

Recently, Mark and co-workers also reported on organophilic silica formed by the combination of the sol-gel procedure and water-in-oil micro-emulsion method, in which methacryloyloxypropyltrimethoxysilane was used as one component of silica matrix [8]. The size of the silica particle was controlled by the content of water and emulsifier used. The surface of the particles was effectively covered with methacryloyl. organic groups. This organophilic silica is expected to be used as a novel component of composite materials. 

The second class of materials, which we will consider herein are carbons with a highly ordered porosity prepared by a template technique [15-18]. The pores are characterized by a well-defined size determined by the wall thickness of the silica substrate used as substrate for carbon infiltration. They can be also interconnected, that is very useful for the charge diffusion in the electrodes. Figure 1 presents the general principle of the carbon preparation by a template technique, where the silica matrix can be, for example, MCM-48 or SBA-15. 

The pores of the silica template can be filled by carbon from a gas or a liquid phase. One may consider an insertion of pyrolytic carbon from the thermal decomposition of propylene or by an aqueous solution of sucrose, which after elimination of water requires a carbonization step at 900°C. The carbon infiltration is followed by the dissolution of silica by HF. The main attribute of template carbons is their well sized pores defined by the wall thickness of the silica matrix. Application of such highly ordered materials allows an exact screening of pores adapted for efficient charging of the electrical double layer. The electrochemical performance of capacitor electrodes prepared from the various template carbons have been determined and are tentatively correlated with their structural and microtextural characteristics. 

Similarly to the above-mentioned entrapment of proteins by biomimetic routes, the sol-gel procedure is a useful method for the encapsulation of enzymes and other biological material due to the mild conditions required for the preparation of the silica networks [54,55]. The confinement of the enzyme in the pores of the silica matrix preserves its catalytic activity, since it prevents irreversible structural deformations in the biomolecule. The silica matrix may exert a protective effect against enzyme denaturation even under harsh conditions, as recently reported by Frenkel-Mullerad and Avnir [56] for physically trapped phosphatase enzymes within silica matrices (Figure 1.3). A wide number of organoalkoxy- and alkoxy-silanes have been employed for this purpose, as extensively reviewed by Gill and Ballesteros [57], and the resulting materials have been applied in the construction of optical and electrochemical biosensor devices. Optimization of the sol-gel process is required to prevent denaturation of encapsulated enzymes. Alcohol released during the... 

Fig. 1.3 Schematic representation of the entrapped enzyme in a silica matrix (left side). Enzymatic activity, under extreme alkaline conditions, of acid phosphatase (A) immobilized in silica sol-gel matrices with or without CTAB, or (B) in solution. Reprinted with permission from [56]. Copyright 2005, American Chemical Society.

The first belief in the possibility of enzyme stabilization on a silica matrix was stated by Dickey in 1955, but he did not give experimental evidence, only mentioning that his experiments were unsuccessful [65]. A sol-gel procedure for enzyme immobilization in silica was first developed by Johnson and Whateley in 1971 [66]. The entrapped trypsin retained about 34 % of its tryptic activity observed in solution before the encapsulation. Furthermore, the enzyme was not released from the silica matrix by washing, demonstrating the increased stability and working pH range. Unfortunately, the article did not attract attention, although their method contained all the details that may be found in the present-day common approach. This was probably due to its publication in a colloid journal that was not read by biochemists. 

Interest in sol-gel processing was awakened by the work of Avnir et al. in 1990 who performed successful experiments with such enzymes as [1-glucosidasc, alkaline phosphatase, chitinase and aspartase [68]. This gave impetus to their own systematic study of the entrapment of biopolymers in a silica matrix as well as those of other teams [69-79]. The results have been summarized and discussed in numerous review articles (see, e.g., Refs. [41—43,45—49,51,80—85]). 

Fig. 3.3 The common two-stage sol-gel process used to entrap biopolymers in a silica matrix (see Scheme 3.1). The first stage serves to hydrolyze alkoxide Equation (2) in the acidic or alkaline media. This is also attended with condensation reactions Equations (3) and (4) resulting in the formation of oligomeric silica that self-organizes in the form of sol nanoparticles. Biopolymers are entrapped in the...

It should be pointed out that the addition of substances, which could improve the biocompatibility of sol-gel processing and the functional characteristics of the silica matrix, is practiced rather widely. Polyethylene glycol) is one of such additives [110— 113]. Enzyme stabilization was favored by formation of polyelectrolyte complexes with polymers. For example, an increase in the lactate oxidase and glycolate oxidase activity and lifetime took place when they were combined with poly(N-vinylimida-zole) and poly(ethyleneimine), respectively, prior to their immobilization [87,114]. To improve the functional efficiency of entrapped horseradish peroxidase, a graft copolymer of polyvinylimidazole and polyvinylpyridine was added [115,116]. As shown in Refs. [117,118], the denaturation of calcium-binding proteins, cod III parvalbumin and oncomodulin, in the course of sol-gel processing could be decreased by complexation with calcium cations. 

These examples demonstrate that additives can have a beneficial effect on the entrapped biopolymers. Unfortunately, they are generally not universal. The additives need to be found for individual immobilized biopolymers and that is not so easy to do. For instance, lactate oxidase retained its activity in a silica matrix if the enzyme was taken as a complex with poly(N-vinylimidazole) prior to the immobilization, but the polymer did not stabilize glycolate oxidase [87,114], Its stabilization was observed after an exchange of poly(N-vinylimidazole) for poly(ethyleneimine). This is a decisive disadvantage of the approaches because they do not offer a general solution that might be extended to any immobilized biopolymer. 

An example of the appropriate application of organically-modified silica precursors is alkoxides with an alkyl group. When methyltrimethoxy- or methyl-triethoxysilane (Figure 3.2) was added in formulations to increase the hydro-phobicity of ORMOSILs, it resulted in a better enzymatic activity of lipases immobilized in the alkyl-modified silica than in a hydrophilic matrix fabricated by means ofTEOS alone [51,80,129-133]. Similarly, an increased stability of lipase from Candida antarctica B was observed after its immobilization in a silica matrix... 

Fig. 3.10 Relative activities of endo-l,3-P-D-glucanase L V in aqueous solution (1) and in the immobilized state (2) as well as immobilized endo-l,3-P-D-glucanase L0 (3) vs. the time of testing. The enzyme entrapment was performed as described in Refs. [55,56], The silica matrix...

The decreased denaturating action of the precursor and procedure enables one to immobilize reduced amounts of biomaterial. It was demonstrated in Ref. [55] that biocatalysts prepared by entrapping endo-l,3-P-D-glucanase and a-D-galactosidasc in amounts comparable to that in living cells had a reasonable level of activity. When the TEOS is applied, the enzyme content in silica matrix can be up to 20-30 wt.% to counterbalance losses due to denaturation [50]. 

The porosity of hybrid nanocomposites provides access of the substrates to immobilized enzyme and their proper functioning. It is attributable to the absence of volume shrinkage of synthesized materials after their preparation. Although the compacting does not occur as in the common two-stage processes (Figure 3.7), enzyme macromolecules are held inside the silica matrix and not easily washed out of it. 

Ferrer, M.L., Yuste, L., Rojo, F. and Del Monte, F. (2003) Biocompatible sol-gel route for encapsulation of living bacteria in organically modified silica matrixes. Chemistry of Materials 15, 3614-3618. 

Given that the primary step in the formation of NPSs is the infiltration of macromolecules into the porous silica matrix, the polymer size is an important parameter. Low molecular weight PEs are preferred for infiltration into the pores of the APTS-BMS spheres, and this process can be promoted by tuning the adsorption... 

Luo, T.J.M., Soong, R., Lane, E., Dunn, B. and Montemagno, C. (2005) Photo-induced proton gradients and ATP biosynthesis produced by vesicles encapsulated in a silica matrix. Nature Materials, 4, 220-224. 

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