Pesticides enzymes

Following work by the same group addressed some of the major problems arising when electrochemical biosensors are in contact with food matrices pH effect and particle effect. Both problems were solved treating the biosensor surface with a Tween20 /phosphate buffer solution (pH 7.5) after the incubation with pesticide. The treatment was successful in removing the particulate, the correct pH for enzyme activity measurement was attained and the pesticide enzyme inhibition... 

Because of the speciflcity of the antibodies, every group of pesticides will require a specific antibody and the corresponding pesticide-enzyme conjugate. 

Phosphorescence can be found with pesticides, enzymes and aromatic hydrocarbons. 

Substances other than enzymes can be immobilized. Examples include the fixing of heparin on polytetrafluoroethylene with the aid of PEI (424), the controUed release of pesticides which are bound to PEI (425), and the inhibition of herbicide suspensions by addition of PEI (426). The uptake of anionic dyes by fabric or paper is improved if the paper is first catonized with PEI (427). In addition, PEI is able to absorb odorizing substances such as fatty acids and aldehydes. Because of its high molecular weight, PEI can be used in cosmetics and body care products, as weU as in industrial elimination of odors, such as the improvement of ambient air quaHty in sewage treatment plants (428). 

Experimental procedures have been described in which the desired reactions have been carried out either by whole microbial cells or by enzymes (1—3). These involve carbohydrates (qv) (4,5) steroids (qv), sterols, and bile acids (6—11) nonsteroid cycHc compounds (12) ahcycHc and alkane hydroxylations (13—16) alkaloids (7,17,18) various pharmaceuticals (qv) (19—21), including antibiotics (19—24) and miscellaneous natural products (25—27). Reviews of the microbial oxidation of aUphatic and aromatic hydrocarbons (qv) (28), monoterpenes (29,30), pesticides (qv) (31,32), lignin (qv) (33,34), flavors and fragrances (35), and other organic molecules (8,12,36,37) have been pubflshed (see Enzyp applications, industrial Enzyt s in organic synthesis Elavors AND spices). 

P. A. Cunniff, ed.. Official Methods of Analysis of AO AC International, 16th ed., Vols. I and II, AO AC International, Arlington, Va., 1995. Vol. I includes Pesticide Formulations and Pesticide Residues. Over 2100 coUabotatively tested, approved methods for chemical and microbiological analyses, with 149 new methods, 103 revised/updated methods, methods using anibody-based test kits, enzyme immunoassay, and annual supplements containing new and revised methods chemical and common names of all dmgs and pesticides easy-to-locate references. 

Pesticides are susceptible to a variety of transformations in the environment, including both chemical degradation and microbial metaboHsm. Microbial transformations are catalyzed exclusively by enzymes, whereas chemical transformations are mediated by a variety of organic and inorganic compounds. Many pesticide transformations can occur either chemically or biologically. Consequentiy, most pesticide dissipation studies include sterile treatments to... 

Oxidative Reactions. The majority of pesticides, or pesticide products, are susceptible to some form of attack by oxidative enzymes. For more persistent pesticides, oxidation is frequently the primary mode of metaboHsm, although there are important exceptions, eg, DDT. For less persistent pesticides, oxidation may play a relatively minor role, or be the first reaction ia a metaboHc pathway. Oxidation generally results ia degradation of the parent molecule. However, attack by certain oxidative enzymes (phenol oxidases) can result ia the condensation or polymerization of the parent molecules this phenomenon is referred to as oxidative coupling (16). Examples of some important oxidative reactions are ether cleavage, alkyl-hydroxylation, aryl-hydroxylation, AJ-dealkylation, and sulfoxidation. 

L-Tyrosine metabohsm and catecholamine biosynthesis occur largely in the brain, central nervous tissue, and endocrine system, which have large pools of L-ascorbic acid (128). Catecholamine, a neurotransmitter, is the precursor in the formation of dopamine, which is converted to noradrenaline and adrenaline. The precise role of ascorbic acid has not been completely understood. Ascorbic acid has important biochemical functions with various hydroxylase enzymes in steroid, dmg, andhpid metabohsm. The cytochrome P-450 oxidase catalyzes the conversion of cholesterol to bUe acids and the detoxification process of aromatic dmgs and other xenobiotics, eg, carcinogens, poUutants, and pesticides, in the body (129). The effects of L-ascorbic acid on histamine metabohsm related to scurvy and anaphylactic shock have been investigated (130). Another ceUular reaction involving ascorbic acid is the conversion of folate to tetrahydrofolate. Ascorbic acid has many biochemical functions which affect the immune system of the body (131). 

Biosensors ai e widely used to the detection of hazardous contaminants in foodstuffs, soil and fresh waters. Due to high sensitivity, simple design, low cost and real-time measurement mode biosensors ai e considered as an alternative to conventional analytical techniques, e.g. GC or HPLC. Although the sensitivity and selectivity of contaminant detection is mainly determined by a biological component, i.e. enzyme or antibodies, the biosensor performance can be efficiently controlled by the optimization of its assembly and working conditions. In this report, the prospects to the improvement of pesticide detection with cholinesterase sensors based on modified screen-printed electrodes are summarized. The following opportunities for the controlled improvement of analytical characteristics of anticholinesterase pesticides ai e discussed ... 

The commercial exploitation of our increased understanding of protein stmcture will not, of course, be restricted to the pharmaceutical industry. The industrial use of enzymes in the chemical industry, the development of new and more specific pesticides and herbicides, the modification of enzymes in order to change the composition of plant oils and plant carbohydrates are all examples of other commercial developments that depend, in part, on understanding the structure of particular proteins at high resolution. 

Chemicals are ubiquitous as air, carbohydrates, enzymes, lipids, minerals, proteins, vitamins, water, and wood. Naturally occurring chemicals are supplemented by man-made substances. There are about 70000 chemicals in use with another 500-1000 added each year. Their properties have been harnessed to enhance the quality of life, e.g. cosmetics, detergents, energy fuels, explosives, fertilizers, foods and drinks, glass, metals, paints, paper, pesticides, pharmaceuticals, plastics, rubber, solvents, textiles thus chemicals are found in virtually all workplaces. Besides the benefits, chemicals also pose dangers to man and the environment. For example ... 

Pesticide inhibition on an active enzyme has been reported, which caused enzyme activities to reduce. The collected data with and without inhibition are presented hi Table E.5.1. Determine the rate model with and without inhibitor (see Table E.5.1). Also define the type of inhibition. 

Toxin (Enzyme Inhibition) Biosensors Enzyme affectors (inhibitors and activators) that influence the rate of biocatalytic reactions can also be measured. Sensing probes for organophosphate and carbamate pesticides, for the respiratory... 

Example 6-2 The following standard addition plot was obtained for a competitive electrochemical enzyme immunoassay of the pesticide 2,4-D. A ground water sample (diluted 1 20 was subsequently assayed by the same protocol to yield a current signal of 65 nA. Calculate the concentration of 2,4-D in the original sample. 

Suggest an enzyme electrode-based procedure for detecting organophosphate pesticides. 

Compounds that affect activities of hepatic microsomal enzymes can antagonize the effects of methyl parathion, presumably by decreasing metabolism of methyl parathion to methyl paraoxon or enhancing degradation to relatively nontoxic metabolites. For example, pretreatment with phenobarbital protected rats from methyl parathion s cholinergic effects (Murphy 1980) and reduced inhibition of acetylcholinesterase activity in the rat brain (Tvede et al. 1989). Phenobarbital pretreatment prevented lethality from methyl parathion in mice compared to saline-pretreated controls (Sultatos 1987). Pretreatment of rats with two other pesticides, chlordecone or mirex, also reduced inhibition of brain acetylcholinesterase activity in rats dosed with methyl parathion (2.5 mg/kg intraperitoneally), while pretreatment with the herbicide linuron decreased acetylcholine brain levels below those found with methyl parathion treatment alone (Tvede et al. 1989). 

Golbs S, Fuchs V, Leipner E, et al. 1978c. [Studies into effects of pesticide combinations in laboratory rats. 3rd communication Experiments about influence on different serum enzymes]. Arch Exp Vet Med Leipzig 32 569-577. (German)... 

Agarwal et al. 1978), the quantification of these specific enzymes may indicate that exposure to endosulfan has occurred. Blood tests, such as decay curves for aminopyrine in plasma, which are semiquantitative indices of liver enzyme induction, have been used successfully in the past to demonstrate enzyme induction in pesticide-exposed workers. Because numerous chemicals found at hazardous waste sites also induce these hepatic enzymes, these measurements are not specific for endosulfan exposure. However, measurements of enzyme activity, together with the detection of the parent compound or its metabolites in tissue or excreta, can be useful indicators of exposure. All of these potential biomarkers require further verification in epidemiological studies. Further studies with focus on the development of methods to separate and measure the estrogenicity of endosulfan in in vitro assays would be valuable since these assays are more sensitive and discriminative than other conventional biomarkers. Preliminary results have been presented by Sonnenschein et al. (1995). 

Den Tonkelaar EM, Van Esch GJ. 1974. No-effect levels of organochlorine pesticides based on induction of microsomal liver enzymes in short-term toxicity experiments. Toxicology 2 371-380. 

Patel Y, Kushwah HS, Kushwah A, et al. 1998a. In Vitro action of pesticides mixture on certain metabolic enzymes in rats. Indian Vet J 75 600-603. 

Siddiqui MKJ, Rahman MF, Anjum F, et al. 1987b. Effect of oral administration of endosulfan on some hematological parameters and serum enzymes in rats. Pesticides 21 25-27. 

Selective toxicity is also important in relation to the development of resistance or tolerance to pollutants from two distinct points of view. On the one hand, there is interest among scientists concerned with crop protection and disease control in mechanisms by which crop pests, vectors of disease, plant pathogens, and weeds develop resistance to pesticides. Understanding the mechanism should point to ways of overcoming resistance, for example, other compounds not affected by resistance mechanisms or synergists to inhibit enzymes that provide a resistance mechanism. On the other hand, the development of resistance can be a useful indication of the environmental impact of pollutants. 

Other interesting examples of proteases that exhibit promiscuous behavior are proline dipeptidase from Alteromonas sp. JD6.5, whose original activity is to cleave a dipeptide bond with a prolyl residue at the carboxy terminus [121, 122] and aminopeptidase P (AMPP) from E. coli, which is a prohne-specific peptidase that catalyzes the hydrolysis of N-terminal peptide bonds containing a proline residue [123, 124]. Both enzymes exhibit phosphotriesterase activity. This means that they are capable of catalyzing the reaction that does not exist in nature. It is of particular importance, since they can hydrolyze unnatural substrates - triesters of phosphoric acid and diesters of phosphonic acids - such as organophosphorus pesticides or organophosphoms warfare agents (Scheme 5.25) [125]. 

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