Immobilization systems cross-linking

A thermally stable NHase from Comamonas testosteroni 5-MGAM-4D (ATCC 55 744) [22] was recombinantly expressed in Escherichia coli, and the resulting transformant cells immobilized in alginate beads that were subsequently chemically cross-linked with glutaraldehyde and polyethylenimine. This immobilized cell catalyst (at 0.5 % dew per reaction volume) was added to an aqueous reaction mixture containing 32wt% 3-cyanopyridine at 25 °C, and a quantitative conversion to nicotinamide was obtained. The versatility of this catalyst system was further illustrated by a systematic study of substrates, which included... 

Typical systems. A considerable number of immobilized polyether systems have been synthesized both for phase transfer catalysis as just discussed and for use in a number of analytical applications. Such immobilized systems are generally synthesized by either copolymerization of suitably functionalized macrocycles in the presence of cross-linking agents or by appending functionalized macrocycles to existing polymeric substrates. Structures (184)-(186) give examples of different... 

Starting from the corresponding hydroxymethyl-benzocrown, it has been possible to generate the immobilized system (186) by reacting the above precursor with chloromethylated polystyrene (which is available commercially as Merrifield s resin). Typically, systems of this type contain a polystyrene matrix which has been cross-linked with approximately 1-4% p-divinylbenzene. In one study involving (186), a clean resolution of the alkali metal halides was achieved by HPLC using (186) as the solid phase and methanol as eluent (Blasius etal., 1980). In other studies, the divalent alkaline earths were also separated. 

Immobilized cryptates. Like the crowns, cryptates have been immobilized on polymeric backbones. A typical system is given by (221) (Cinquini, Colonna, Molinari, Montanari Tundo, 1976). In this case, the polymeric matrix is polystyrene cross-linked with p-divinyl benzene and the cage is connected to this matrix via a long-chain aliphatic spacer group. This reagent is quite effective as a (triphase) transfer catalyst. 

With a view to producing catalysts that can easily be removed from reaction products, typical phase-transfer catalysts such as onium salts, crown ethers, and cryptands have been immobilized on polymer supports. The use of such catalysts in liquid-liquid and liquid-solid two-phase systems has been described as triphase catalysis (Regen, 1975, 1977). Cinquini et al. (1976) have compared the activities of catalysts consisting of ligands bound to chloromethylated polystyrene cross-linked with 2 or 4% divinylbenzene and having different densities of catalytic sites ([126], [127], [ 132]—[ 135]) in the... 

Fig. 16. Cross-relaxation or Z-spectra derived from the solvent proton spectra of cross-linked bovine serum albumin gel supported in a ternary solvent system consisting of 8.8% H2O, 8.7% acetone, and 8.8% methanol and 63% D20.The offset axis represents the frequency offset of the off-resonance preparation pulse which partly saturates the immobilized spin. In this case the 3 s preparation had an amplitude of 880 Hz at a Larmor frequency of 500 MHz (87).

Another coupling method, i.e. cross-linking or entrapment in polymeric films, which has been used to create a more permanent nucleic acid surface, is described in some chapters (e.g. conductive electroactive polymers for DNA immobilization and self-assembly DNA-conjugated polymers). One chapter reviews the basic characteristics of the biotin-(strept)avidin system laying the emphasis on nucleic acids apphcations. The biotin-(strept)avidin system can be also used for rapid prototyping to test a large number of protocols and... 

Schmid et al. used the same principle to develop sensors to be incorporated into FI systems for the determination of ascorbic acid in fruit juices [38] and that of lactic acid in dairy products [39]. The membrane used in both applications consisted of decacyclene dissolved in silicone rubber that was treated similarly as the membrane in glucose sensors (Fig. 3.4.B). The oxygen optrode was coated with a sheet of carbon black as optical insulation in order to protect it from ambient light or intrinsic sample fluorescence. Ascorbic acid oxidase or lactic acid oxidase was immobilized by adsorbing it onto carbon black and cross-linking it with glutaraldehyde. The FI system automatically buffered and diluted the food samples, thereby protecting the biosensor from a low pH and interferents. 

The data presented in Figure 4.14a are consistent with the following mechanism. The dispersion that emerges from the blender is fundamentally unstable with respect to coagulation and coagulates rapidly to form a volume-filling network throughout the continuous phase. Except for the size and structure of the chains, the situation is comparable to a cross-linked polymer swollen by solvent. In both, the liquid is essentially immobilized by the network of chains, and the system behaves as an elastic solid under low stress. The term gel is used to describe such systems whether the dispersed particles are lyophilic or lyophobic. 

Kokufuta, Zhang and Tanaka developed a gel system that undergoes reversible swelling and collapsing changes in response to saccharides, sodium salt of dextran sulfate (DSS) and a-methyl-D-mannopyranoside (MP) [126]. The gel consists of a covalently cross-linked polymer network of W-isopropylacrylamide into which concanavalin A (ConA) is immobilized. As shown in Fig. 31, at a certain temperature the gel swells five times when DSS ions bind to ConA due to the excess ionic pressure created by DSS. The replacement of the DSS by non-ionic MP brings about collapse of the gel. The transition can be repeated with excellent reproducibility. 

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