Enzymes pH and

The reason is that the minerals serve as a shield physically protecting cellulose fibers from cellulolytic enzymes diffusing into the matrix. At pH 7.5-9, the enzymes have 10-1000 times lower activity, depending on the types of the enzymes, pH, and the dissociation constants of catalytically active groups in the enzyme active center. 

Giving the various compounds that can be expressed differently in the plant, the relationship between the best solvent to be used with a plant can be complicated by the part of plant (leave, stem, root, or bark of these parts) to be extracted from and this was reported in several studies [133-136]. Several other factors need to be considered during the extraction and evaluation of the compounds such as the distribution of the compounds in the plant, extraction system (solvent, time, temperature, and solid liquid ratio), the effects of storage, chemicals used, dialysis, use of enzymes, pH, and centrifugation. 

Firefly. Firefly luciferase (EC 1.13.12.7) is a homodimeric enzyme (62 kDa subunit) that has binding sites for firefly luciferin and Mg ATP . Amino acid sequence analysis has iadicated that beetle luciferases evolved from coen2yme A synthetase (206). Firefly bioluminescence is the most efficient bioluminescent reaction known, with Qc reported to be 88% (5), and at 562 nm (56). At low pH and ia the presence of certain metal ions (eg, Pb ", ... 

Assays using equiUbrium (end point) methods are easy to do but the time requited to reach the end point must be considered. Substrate(s) to be measured reacts with co-enzyme or co-reactant (C) to produce products (P and Q) in an enzyme-catalyzed reaction. The greater the consumption of S, the more accurate the results. The consumption of S depends on the initial concentration of C relative to S and the equiUbrium constant of the reaction. A change in absorbance is usually monitored. Changes in pH and temperature may alter the equiUbrium constant but no serious errors are introduced unless the equihbrium constant is small. In order to complete an assay in a reasonable time, for example several minutes, the amount and therefore the cost of the enzyme and co-factor maybe relatively high. Sophisticated equipment is not requited, however. 

Enzyme Assays. An enzyme assay determines the amount of enzyme present in sample. However, enzymes are usually not measured on a stoichiometric basis. Enzyme activity is usually determined from a rate assay and expressed in activity units. As mentioned above, a change in temperature, pH, and/or substrate concentration affects the reaction velocity. These parameters must therefore be carefully controlled in order to achieve reproducible results. 

Enzyme Sta.bihty, Loss of enzyme-catalytic activity may be caused by physical denaturation, eg, high temperature, drying/freezing, etc or by chemical denaturation, eg, acidic or alkaline hydrolysis, proteolysis, oxidation, denaturants such as surfactants or solvents, etc. pH has a strong influence on enzyme stabiHty, and must be adjusted to a range suitable for the particular enzyme. If the enzyme is not sufficiendy stable in aqueous solution, it can be stabilized by certain additives a comprehensive treatment with additional examples is available (27). 

A given enzyme may be assayed by its action on soluble substrates under chemical and physical conditions different from those encountered in a real-life wash. Such experiments indicate the enzyme s performance with respect to pH and temperature variations, or in conjunction with other soluble substances, etc. The analytical data thus obtained are not necessarily representative of the wash performance of the enzyme, and real wash trials are necessary to evaluate wash performance of detergent enzymes. 

Most ingredients in a detergent formulation contribute to the ionic strength of the wash solution. The effect of ionic strength on protease performance depends on pH and enzyme identity. The pH wash solutions also affects protease performance (Pig. 8). 

When selecting a suitable feed symp, the main criteria are optimization of enzyme productivity and minimization of the formation of by-products. Typical feed symp specifications are shown in Table 5. Higher symp concentration and higher viscosity results in a reduced isomerization rate due to diffusion resistance in the pores of the immobilized enzyme. A deaeration step is desirable to remove dissolved oxygen that would otherwise iacrease the formation of by-products. The pH is adjusted to the optimum level for the productivity of the enzyme. 

Enzyme reac tions are also sensitive to pH and temperature changes. In characterizing an enzyme, its optimal pH and optimal temperature are conditions at which the enzyme has its highest catalytic activity. 

Figure 18.4 The hanging-drop method of protein crystallization, (a) About 10 pi of a 10 mg/ml protein solution in a buffer with added precipitant—such as ammonium sulfate, at a concentration below that at which it causes the protein to precipitate—is put on a thin glass plate that is sealed upside down on the top of a small container. In the container there is about 1 ml of concentrated precipitant solution. Equilibrium between the drop and the container is slowly reached through vapor diffusion, the precipitant concentration in the drop is increased by loss of water to the reservoir, and once the saturation point is reached the protein slowly comes out of solution. If other conditions such as pH and temperature are right, protein crystals will occur in the drop, (b) Crystals of recombinant enzyme RuBisCo from Anacystis nidulans formed by the hanging-drop method. (Courtesy of Janet Newman, Uppsala, who produced these crystals.)...

Mammals, fungi, and higher plants produce a family of proteolytic enzymes known as aspartic proteases. These enzymes are active at acidic (or sometimes neutral) pH, and each possesses two aspartic acid residues at the active site. Aspartic proteases carry out a variety of functions (Table 16.3), including digestion pepsin and ehymosin), lysosomal protein degradation eathepsin D and E), and regulation of blood pressure renin is an aspartic protease involved in the production of an otensin, a hormone that stimulates smooth muscle contraction and reduces excretion of salts and fluid). The aspartic proteases display a variety of substrate specificities, but normally they are most active in the cleavage of peptide bonds between two hydrophobic amino acid residues. The preferred substrates of pepsin, for example, contain aromatic residues on both sides of the peptide bond to be cleaved. 

The latter would normally require a high pH and the contribution of the enzyme is therefore presumed to be the provision of a suitable environment, within the protein pocket, which allows the dissociation of the coordinated H,0 to occur in a medium of pH 7 which would otherwise be much too low. 

Enzyme activity in organic solvents depends on parameters such as water activity, pH control, substrate-product solvation, enzyme form, and nature of the solvent. 

When discussing the role of reaction medium on enzyme enantioselectivity, the potential effects of (i) water activity [5b,13f,32], (ii) enzyme form, and (iii) pH, should... 

Biocatalysts usually require mild reaction conditions for an optimal activity (physiologic temperature and pH) and, in general, they show high activity, chemo- and enantioselectivity. Furthermore, when using enzymes, many functional group protections and/or activations can be avoided, allowing shorter synthetic transformations. The use of enzymes is therefore very attractive from an environmental and economic point of view. 

The protected E-ring moiety of (S)-camptothecin has been prepared in enantio-merically enriched form by the enzymatic resolution of a triester with PLE in a pH 7 phosphate buffer-acetonitrile (5 1) solution (Figure 6.7). The alkaloid camptothecin is an inhibitor of the enzyme topoisomerase and some of its derivatives are anticancer drugs [52]. 

O2, Mn, pH, and solid concentrations indicates that the character of the solid is important partly because some surfaces bind Mn " more strongly and partly because they facilitate the electron transfer differently. Catalysis by enzymes is clearly the most effective oxidation enhancing process as indicated by the laboratory studies with spores and material from the O2/H2S interface of Saanich Inlet. Microbial catalysis in this environment reduces the oxidation lifetime of Mn to about one day. This example illustrates... 

Table 16-7 The surface or enzyme-catalyzed reaction rate constant, Mn/ for oxidation of Mn normalized for oxygen concentration [O2], pH and particulate concentration [X]. d[Mn ]/dt = Mn [Mn][02][0H] [X]...

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