Gel beads

Because enzymes can be intraceUularly associated with cell membranes, whole microbial cells, viable or nonviable, can be used to exploit the activity of one or more types of enzyme and cofactor regeneration, eg, alcohol production from sugar with yeast cells. Viable cells may be further stabilized by entrapment in aqueous gel beads or attached to the surface of spherical particles. Otherwise cells are usually homogenized and cross-linked with glutaraldehyde [111-30-8] to form an insoluble yet penetrable matrix. This is the method upon which the principal industrial appHcations of immobilized enzymes is based. 

The presence of a COOH functionality on a polystyrene resin can be detected using a 0.25% solution of malachite green-oxalate in ethanol in the presence of a drop of triethylamine. Beads with COOH functionalities are coloured dark green or appear as clear gel beads [Attardi, Porcu and Taddei Tetrahedron Lett 41 7391 2000]. 

FIGURE 5A.1 A gel filtration chromatogr phy column. Larger molecules are excluded from the gel beads and emerge from the column sooner than smaller molecules, whose migration is retarded because they can enter the beads. 

Wang et al.2 and Najafpour et al.3A worked with immobilised microbial cells of Nitrobacer agilis, Saccharomyces cerevisiae and Pseudomonas aeruginosa in gel beads, respectively. They found separately that the cells retained more than 90% of their activity after immobilisation by using specific oxygen uptake rate (SOUR) [mg 02g 1 (dry biomass) h 11 as the biomass activity indicator. Such differences in immobilised biomass and activity between free and immobilised biomass activities depend strongly on the particular characteristics of the microbial systems and their interaction with the support matrix. 

The separation on gel beads was more and more refined. Oligomer series of many polyhexapeptides from n = lton = 9 could be completely separated16,20 24) (see Fig. 4). The hexapeptide oligomers could be separated better than the respective tripeptides. 

A simple example of gel formation is provided by chitosan tripolyphosphate and chitosan polyphosphate gel beads the pH-responsive swelling abihty, drug-release characteristics, and morphology of the gel bead depend on polyelectrolyte complexation mechanism and the molecular weight. The chitosan beads gelled in pentasodium tripolyphosphate or polyphosphoric acid solution by ionotropic cross-hnking or interpolymer complexation, respectively. 

Direct measurement of adsorptive stripping voltaimnetric peaks using HMDE 0.60 V and accumulation potential of -0.40V Dilution in phosphate buffer and water, analyzed in Vis region Ion pair formation with octadecyltrimethylammonium bromide at pH 5.6, extraction of ion pair into n-butanol Sample solution mixed with 1 M HCl, ethanol and purification on Sephadex DEAE 25 gel, gel beads are filtered off, packed into 1 nun cell and absorbance measured... 

At end, it is important to mention that calcium pectate gel beads were compared with calcium alginates gel beads for all entrapment uses [65, 66] in this work, the authors determined the pore size of the beads by size exclusion chromatography using dextran standards and other solutes. 

C-J Kim, PI Lee. Hydrophobic anionic gel beads for swelling-controlled drug delivery. Pharm Res 9 195-201, 1991. 

Scheme 5.7 Encapsulation of enzyme microcapsules into a gel-like structure (host gel bead) resulting in a capsules-in-bead microreactor. Reproduced from [20] by permission of The Royal Society of Chemistry.

Multiple-enzyme conversion of CO2 to formate, then formaldehyde, then methanol by FateDH, FaldDH, and ADH Enzymes loaded in CaC03 followed by LbL deposition and dissolution of core, then encapsulation into a gel bead... 

The effectiveness of zeolites in catalysis and separation can often be improved by the textural and chemical properties of the matrices in which they are imbedded. Chitosan gels issued from renewable resources are already used as supports for the preparation of heterogeneous catalysts in the form of colloids, flakes or gel beads [1, 2], In this study we present several methods for the incorporation of zeolites in chitosan matrices and characterize the synergic effect of the components on the properties of the composite. 

Chen et al. [54] have reported a model for the assessment of the combined effects of the intrinsic reaction kinetics and dye diffusion into phosphorylated polyvinyl alcohol (PVA) gel beads. The analysis of the experimental data in terms of biofilm effectiveness factor highlighted the relevance of intraparticle diffusion to the effective azo-dye conversion rate. On the basis of these results, they have identified the optimal conditions for the gel bead diameter and PVA composition to limit diffusion resistance. 

Yoshimura et al. [193] carried out microdeterminations of phosphate by gel-phase colorimetry with molybdenum blue. In this method phosphate reacted with molybdate in acidic conditions to produce 12-phosphomolybdate. The blue species of phosphomolybdate were reduced by ascorbic acid in the presence of antimonyl ions and adsorbed on to Sephadex G-25 gel beads. Attenuation at 836 and 416 nm (adsorption maximum and minimum wavelengths) was measured, and the difference was used to determine trace levels of phosphate. The effect of nitrate, sulfate, silicic acid, arsenate, aluminium, titanium, iron, manganese, copper, and humic acid on the determination were examined. 

Columns of gel beads are more often used than thin layers. The process can be considered analogous to a partition system wherein molecules which are completely excluded from the gel have a distribution ratio of 0 whilst those small enough to penetrate all parts of the structure have a value of 1. Adsorption or ion-exchange effects can cause the distribution ratios of polar molecules to exceed 1. Components of a mixture are characterized by their retention volume, VR, which is determined by the distribution ratio. 

Relative molecular mass distributions for components of biochemical and polymer systems can be determined with a 10% accuracy using standards. With biochemical materials, where both simple and macro-molecules may be present in an electrolyte solution, desalting is commonly employed to isolate the macromolecules. Inorganic salts and small molecules are eluted well after such materials as peptides, proteins, enzymes and viruses. Desalting is most efficient if gels with relatively small pores are used, the process being more rapid than dialysis. Dilute solutions of macro-molecules can be concentrated and isolated by adding dry gel beads to absorb the solvent and low RMM solutes. 

Microbes tend to form flocks as they grow, into which nutrients and dissolved oxygen must diffuse. The rate of growth thus depends on the diffusional effectiveness. This topic is developed by Atkinson (1974). Similarly enzymes immobilized in gel beads, for instance, have a reduced catalytic effectiveness analogous to that of porous granular catalysts that are studied in Chapter 7. For the M-M equation this topic is touched on in problems P8.04.15 and P8.04.16. 

Yang, Q., Liu, X. Y., Umetani, K., Kamo, N. and Miyake, J. (1999). Partitioning of triphenylalkylphosphonium homologues in gel bead-immobilized liposomes chromatographic measurement of their membrane partition coefficients, Biochim. Biophys. Acta-Biomem., 1417, 122-130. 

Despite the success with gel-phase NMR spectroscopy the spectra obtained are of limited value because of the broad peaks, and indeed the NMR spectra of unlabelled samples need relatively long acquisition times (because of the low abundance of C) especially with lightly loaded resins such as TentaGel and Argo-Gel. NMR [206, 207] and NMR [208, 209] spectroscopy of gel beads have also been shown to be a convenient NMR technique for monitoring reactions of fluorine or phosphorous-containing molecules, respectively, attached to solvent-swollen polymer supports. 

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