Enzymes, detergent compound

National Academy of Science-National Research Council, Division of Medical Science, Report of the ad hoc Committee on Enzyme Detergents Enzyme-Containing Laundering Compounds and Consumer Health, supported by Food and Drug Administration, 1971. 

When binding of a substrate molecule at an enzyme active site promotes substrate binding at other sites, this is called positive homotropic behavior (one of the allosteric interactions). When this co-operative phenomenon is caused by a compound other than the substrate, the behavior is designated as a positive heterotropic response. Equation (6) explains some of the profile of rate constant vs. detergent concentration. Thus, Piszkiewicz claims that micelle-catalyzed reactions can be conceived as models of allosteric enzymes. A major factor which causes the different kinetic behavior [i.e. (4) vs. (5)] will be the hydrophobic nature of substrate. If a substrate molecule does not perturb the micellar structure extensively, the classical formulation of (4) is derived. On the other hand, the allosteric kinetics of (5) will be found if a hydrophobic substrate molecule can induce micellization. 

The digestion and absorption of fat is considerably more complex than that of carbohydrate or protein because it is insoluble in water, whereas almost aU enzymes catalyse reactions in an aqueous medium. In such media, fat can form small droplets, an emulsion, which is stable in this medium. Formation of an emulsion is aided by the presence of detergents these possess hydrophobic and hydrophilic groups, so that they associate with both the fat and the aqueous phases. Such compounds are known as emulsifying agents and those involved in digestion are mainly the bile salts and phospholipids. 

From bisubstrate, kinetic analysis with a transferase from hen oviduct that, under the conditions of the assay, formed only GlcNAc-PP-Dol, it followed that both dolichol phosphate and UDP-GlcNAe have to he bound to the enzyme before release of the product occurs.52 However, the fact that only partially purified preparations have thus far been obtained (the preparations may also still be contaminated with substrates and product), together with experimental difficulties in handling both the substrate dolichol phosphate (which, furthermore, is not one compound, see the earlier discussion) and the unstable enzyme (enveloped in micelles of detergent), make difficult a sensible interpretation and comparison of the kinetic parameters detenuined for the different enzvme-preparations. The solubilized enzymes catalyzing reactions 1,2, and 3 have in common their alkaline pH optima and dependence on Mg2+ or Mn2+ ions. The latter fact makes (ethylenedinitrilo)tetraacetic acid (EDTA) a reversible inhibitor of enzyme activity and an important experimental tool. 

Both the plasma membrane and internal membranes associated with organelles may be damaged by toxic compounds. Chemicals such as detergents, strong acids and alkalies, and snake venoms, which contain hydrolytic enzymes, can also directly damage the plasma membrane. 

For the study of polyprenyl glycosyl phosphates as intermediates in the synthesis of complex glycans, several techniques have been developed, and these have been described elsewhere in detail.18 20 Two important features should be emphasized. First, the very small amounts of polyprenyl phosphates that are present in most tissues for this reason, the use of radioactive techniques for the detection of products is obligatory. Second, on account of the hydrophobic nature of these compounds, and as the enzymes involved in the reactions are membrane-bound, the use of detergents and organic solvents becomes necessary. 

Surfactants, emulsifiers Dissolve lipids, disrupt membranes, denature proteins, and inactivate enzymes in high concentrations, and act as wetting agents Cationic detergents are used to sanitize utensils, anionic detergents are used to launder clothes and clean household objects, and quaternary ammonium compounds are sometimes used as an antiseptic on skin... 

The solubilization phenomenon, which refers to the dissolution of normally insoluble or only slightly soluble compounds in water caused by the addition of surfactants, is one of the most striking effects encountered for surfactant systems. Solubilization is of considerable physico-chemical interst, such as in discussion of the structure and dynamics of micelles and of the mechanism of enzyme catalysis, and has numerous practical applications, such as in detergency, in pharmaceutical preparations and in micellar catalysis. In biology, solubilization phenomena are most significant, e.g., cholesterol solubilization in phospholipid bilayers and fat solubilization in fat digestion and transport. 

We have now extended these studies to synthetic phospholipids that contain short chain fatty acyl groups and which are water soluble, such as dibutyryl and dihexanoyl phosphatidylcholine (PC). These phospholipids are monomeric below their critical micelle concentration (cmc), yet activate the enzyme. In order to carry out kinetic studies, the long chain phospholipid substrate must generally be solubilized by a detergent such as Triton X-100 which serves as an inert matrix. Further understanding of the mechanism of the activation by short-chain phospholipids requires first a quantitation of the solubilization of these compounds by detergent ... 

The compounds with final high rank orders are then tested experimentally using inhibition assays. The number of compounds to test is generally limited by the monetary and time cost of the experimental assays. The assays can then be repeated in the presence of detergent, e.g., Triton X-100, as a first test to filter out promiscuous, aggregate-based inhibitors (46). The inhibitory effect of the active compounds can also be tested on similar enzymes to investigate the specificity of the compounds for the enzyme of interest (see Note 16). 

Surfactants are frequently used in detergents and food products to alter the properties of solution interfaces, mediating between immiscible phases because of their hydrophobic and hydrophilic moieties. The addition of surfactants increases the concentration of hydrophobic compounds in the water phase by solubilization or emulsification above a specific threshold, the critical micellar concentration (CMC), where surfactant molecules aggregate to micelles [130]. Two widely utilized nonionic surfactants, Tween 80 and Triton X-100, were evaluated in terms of enzyme interaction, by calculating the inactivation coefficient (kA) under static conditions. Concentrations lower than CMC were studied in order to avoid diffusional limitations in the interaction of the enzyme and the PAH in the micellar phase. The concentration 0.25 CMC was considered the most favorable for the enzyme, with Triton X-100 being the surfactant that led to the lowest inactivation coefficients for all the concentrations tested and was 2.5 times lower than kd in control experiment. 

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