Liquid-phase reactants enzymes

It is not unusual for the full chemical potential of a reaction to be diminished by slower transport processes (i.e., to be transport limited). In fast liquid phase enzyme reactions, mechanical stirring rates can have a strong influence on the observed kinetics that may be limited by the rate of contacting of the reactants and enzymes. Most heterogeneous catalytic reactions take... 

Different enzymes - particularly lipases - immobilized in membrane reactors have been studied in the presence of two-liquid phases (Table 5). Organic and aqueous phases containing respectively hydrophobic and hydrophilic reactants are separated by a solid mem-... 

Chromatographic fixed-bed reactors consists of a single chromatographic column containing a solid phase on which adsorption and reaction take place. Normally a pulse of reactant is injected into the reactor and, while traveling through the reactor, simultaneous conversion and separation take place (Fig. 3). Since an extensive overview of the models and applications of this type of reactor was presented by Sardin et al. [ 132], only a few recent results will be discussed here. Most of the practical applications have been based on gas-liquid systems, which are not applicable for the enzyme reactions, but a few reactions were also reported in the liquid phase. One of these studies, performed by Mazzotti and co-workers [ 141 ], analyzed the esterification of acetic acid into ethyl acetate according to the reaction ... 

Most liquid phase chemical and biochemical reactions, with or without catalysts or enzymes, can be carried out either batchwise or continuously. For example, if the production scale is not large, then a reaction to produce C from A and B, all of which are soluble in water, can be carried out batchwise in a stirred tank reactor that is, a lank equipped with a mechanical stirrer. The reactants A and B are charged into the reactor at the start of the operation. The product C is subsequently produced from A and B as time goes on, and can be separated from the aqueous solution when its concentration has reached a predetermined value. 

For liquid-phase catalytic or enzymatic reactions, catalysts or enzymes are used as homogeneous solutes in the hquid, or as sohd particles suspended in the hquid phase. In the latter case, (i) the particles per se may be catalysts (ii) the catalysts or enzymes are uniformly distributed within inert particles or (hi) the catalysts or enzymes exist at the surface of pores, inside the particles. In such heterogeneous catalytic or enzymatic systems, a variety of factors that include the mass transfer of reactants and products, heat effects accompanying the reactions, and/or some surface phenomena, may affect the apparent reaction rates. For example, in situation (iii) above, the reactants must move to the catalytic reaction sites within catalyst particles by various mechanisms of diffusion through the pores. In general, the apparent rates of reactions with catalyst or enzymatic particles are lower than the intrinsic reaction rates this is due to the various mass transfer resistances, as is discussed below. 

Investigating the kinetically controlled synthesis of the /flactam antibiotic amoxicillin from 6-aminopenicillanic acid and D-p-hydroxyphenylglycine methyl ester in a solid suspension system in which the reaction nevertheless occurred in the liquid phase, Diender et al. found that the pH value and dissolved concentrations took a very different course at different initial substrate amounts (Diender, 2000). These results were described reasonably well by the model based on mass and charge balances, pH-dependent solubilities of the reactants, and enzyme kinetics. 

Enzymes are proteins that catalyze reactions. Thousands of enzymes have been classified and there is no clear limit as to the number that exists in nature or that can be created artificially. Enzymes have one or more catalytic sites that are similar in principle to the active sites on a solid catalyst that are discussed in Chapter 10, but there are major differences in the nature of the sites and in the nature of the reactions they catalyze. Mass transport to the active site of an enzyme is usually done in the liquid phase. Reaction rates in moles per volume per time are several orders of magnitude lower than rates typical of solid-catalyzed gas reactions. Optimal temperatures for enzymatic reactions span the range typical of living organisms, from about 4°C for cold-water fish, to about 40°C for birds and mammals, to over 100°C for thermophilic bacteria. Enzymatic reactions require very specific molecular orientations before they can proceed. As compensation for the lower reaction rates, enzymatic reactions are highly selective. They often require specific stereoisomers as the reactant (termed the substrate in the jargon of biochemistry) and can generate stereospecific products. Enzymes are subject to inhibition and deactivation like other forms of catalysis. 

One of the expected benefits from using enzymes in supercritical fluids (SCFs) is that mass transfer resistance between the reaction mixture and the active sites in the solid enzyme should be greatly reduced if the reactants and products are dissolved in an SCF instead of running the reaction in a liquid phase. It is expected that the high diffusivity and low viscosity of SCFs will accelerate mass-transfer controlled reactions. 

Enzyme catalysis in nearly solid or semi-solid systems has been thoroughly studied (Ulijn et al. 2003). In this system the reaction mixture consists of solid reactants suspended in a comparatively small volume of liquid phase, either aqueous (van Langen et al. 1999, 2000) or organic (Basso et al. 2000) that becomes saturated reaction ensues and the product formed precipitates out If om that liquid phase. This... 

Traditionally, the field of catalysis is divided into three areas heterogeneous, homogeneous, and enzyme catalysis. Heterogeneous catalysts are present in a phase different from that of the reactants typically, the reactants are in the gas or liquid phase, whereas the catalyst is a solid material. Homogeneous catalysts operate in the same phase as the reactants, and enzyme catalysts are speciahzed proteins. The chemically active part of enzymes is often a tiny part of the protein, and enzyme catalysis can be viewed as a special kind of heterogeneous catalysis. 

The enzyme may be dissolved in a mixed aqueous-ionic liquid medium, which may be mono- or biphasic or it could be suspended or dissolved in an ionic liquid, with little or no water present. Alternatively, whole cells could be suspended in an ionic liquid, in the presence or absence of a water phase. Mixed aqueous-organic media are often used in biotransformations to increase the solubility of hydrophobic reactants and products. Similarly, mixed aqueous-ionic liquid media have been used for a variety of biotransformations, but in most cases there is no clear advantage over water-miscible organic solvents such as tert-butanol. 

Many enzyme reactions in the pharmaceuticals sector are carried out in an aqueous medium, i.e., the enzyme is dissolved in water, and the reactant is usually a second (water-insoluble) phase. Fundamentally, it would be interesting to learn what would happen if the phases were reversed, i.e., if the reactant were dissolved in some solvent and the enzyme the second phase. Klibanov and coworkers had previously reported on enzymatic catalysis reactions in organic liquids with only scant moisture present to render the enzyme active (Zaks and Klibanov, 1984). 

In many cases an attractive option is to use no solvent at all. In some cases at least one of the reactants will be liquid, so can be the basis of a fluid phase for transfer of reactants. If slightly raised temperatures are used, this condition will be met more often. (Remember that at reduced water activity, the enzyme will usually be stable to higher temperatures than in aqueous solution.)... 

In heterogeneous catalysis, which is of great industrial importance, the catalyst is a solid and the reactants are gases or liquids. In homogeneous catalysis, the catalyst and the reactants are in the same phase. Enzymes are catalysts in living systems. 

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