Immobilized enzymes/cells

Figure 3.8 — (A) Biosensors used in different FI manifolds to perform reaction-rate measurements (I) stopped-flow manifold (II) iterative flow-reversal system (III) open-closed configuration S sample B buffer P pump IV injection valve PC personal computer IMEC immobilized enzyme cell D detector W waste SV switching valve. (B) Types of recordings obtained by using the three types of biosensors and measurements to be performed on them in order to develop reaction-rate methods. (Reproduced from [50] with permission of Elsevier Science Publishers).

The principle of the mass transport of substrates/nutrients into the immobilized enzyme/cells, through a solid, porous layer (membrane, biofilm) or through a gel layer of enzyme/cells is the same. The structure, the thickness of this mass-transport layer can be very different, thus, the mass-transport parameters, namely diffusion... 

A sensitized electrode is a composite device that combines a membrane reactor and a primary electrode. The membrane reactor converts a specific analyte into products that are measurable by the primary electrode. Membrane reactors containing immobilized enzymes, cells, or neutral carriers are capable of very selective conversion of sugars, amino acids, organic acids, and alcohols, and... 

Yoda K. 1988. Immobilized enzyme cells. Methods Enzymology, 137 61. 

Yoda K. (1988) Multienzyme membrane electrodes, in Methods in Enzymology, vol. 137. (Immobilized enzymes cells, Pt D), 61-68. 

Biosynthesis ofS(— )-M llc Acid. Aqueous fumaric acid is converted to levorotatory malic acid by the intracellular enzyme, fumarase, which is produced by various microorganisms. A Japanese process for continuous commercial production of S(—)-mahc acid from fumaric acid is based on the use of immobilized Brevibacteriumflavum cells in carrageenan (32). The yield of pyrogen-free S(—)-mahc acid that is suitable for pharmaceutical use is ca 70% of the theoretical. 

Immobilization. Enzymes, as individual water-soluble molecules, are generally efficient catalysts. In biological systems they are predorninandy intracellular or associated with cell membranes, ie, in a type of immobilized state. This enables them to perform their activity in a specific environment, be stored and protected in stable form, take part in multi-enzyme reactions, acquire cofactors, etc. Unfortunately, this optimization of enzyme use and performance in nature may not be directiy transferable to the laboratory. 

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. 

Some of the industrial biocatalysts are nitrile hydralase (Nitto Chemicals), which has a productivity of 50 g acrylamide per litre per hour penicillin G amidase (Smith Kline Beechem and others), which has a productivity of 1 - 2 tonnes 6-APA per kg of the immobilized enzyme glucose isomerase (Novo Nordisk, etc.), which has a productivity of 20 tonnes of high fmctose syrup per kg of immobilized enzyme (Cheetham, 1998). Wandrey et al. (2000) have given an account of industrial biocatalysis past, present, and future. It appears that more than 100 different biotransformations are carried out in industry. In the case of isolated enzymes the cost of enzyme is expected to drop due to an efficient production with genetically engineered microorganisms or higher cells. Rozzell (1999) has discussed myths and realities... 

All soil metabolic proce.sses are driven by enzymes. The main sources of enzymes in soil are roots, animals, and microorganisms the last are considered to be the most important (49). Once enzymes are produced and excreted from microbial cells or from root cells, they face harsh conditions most may be rapidly decomposed by organisms (50), part may be adsorbed onto soil organomineral colloids and possibly protected against microbial degradation (51), and a minor portion may stand active in soil solution (52). The fraction of extracellular enzyme activity of soil, which is not denaturated and/or inactivated through interactions with soil fabric (51), is called naturally stabilized or immobilized. Moreover, it has been hypothesized that immobilized enzymes have a peculiar behavior, for they might not require cofactors for their catalysis. 

Polyamines and their ammonium salts have been of interest because they are known to have potential applications as chelating agents (1-3), ion exchange resins (4-6), flocculants (7,8), and other industrial uses (9). Recent biomedical applications have constituted another important use of polymeric amines they have been investigated for use as biocompatable materials, polymeric drugs, immobilization of enzymes, cell-culture substratum and cancer chemotherapeutic agents (10-12). 

In another approach, the alcohol moiety, formed by an enzymatic hydrolysis of an ester, can act as a nucleophile. In their synthesis of pityol (8-37a), a pheromone of the elm bark beetle, Faber and coworkers [17] used an enzyme-triggered reaction of the diastereomeric mixture of ( )-epoxy ester 8-35 employing an immobilized enzyme preparation (Novo SP 409) or whole lyophilized cells of Rhodococcus erythro-polis NCIMB 11540 (Scheme 8.9). As an intermediate, the enantiopure alcohol 8-36 is formed via kinetic resolution as a mixture ofdiastereomers, which leads to the diastereomeric THF derivatives pityol (8-37a) and 8-37b as a separable mixture with a... 

Dias, A.C.P, Cabral, J.M.S. and Pinheiro, H.M. (1994) Sterol side-chain cleavage with immobilized Mycobacterium cells in water-immiscible organic solvents. Enzyme and Microbial Technology, 16, 708-714. 

Klibanov, A.M. (1983) Immobilized enzymes and cells as practical catalysts. 

Reactions of cell growth or those using immobilized enzymes are instances of gas-liquid-solid reactions. In principle, accordingly, any of the types of reactors described in Section 8.3 could be employed as fermentors. Mostly, however, mechanically agitated tanks are the type adopted. Aeration supplies additional agitation as well as metabolic need, and moreover sweeps away C02 and noxious byproducts. 

Immobilized catalases, for textile bleaching effluent treatment, 4 68—69 Immobilized cells, 3 670 Immobilized enzyme biosensors,... 

Immobilized enzymes and immobilized cells can be used in principle as heterogeneous catalysts not only for the enhancement of hydrolytic reactions but also in specific synthetic reactions. Many of the enzymes involved in the synthesis of complex biological compounds -in vivo and in vitro require cofactors which have to be provided in suitable form when immobilized enzymes or cells are used. 

Immobilized enzymes and immobilized cells are being employed as specific heterogeneous catalysts by several of the chemical industries. An evaluation of the mode of action and efficiency of... 

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