Insect esterase

Methods have been published that allow the classification of two types of esterases, the carboxylic ester hydrolases (CEHs) and the phosphoric triester hydrolases (PTEHs) (Anspaugh and Roe, 2004). The CEHs contain the B-esterases, which are inhibited by organophosphates. B-esierases include many other esterases, such as CarbE, acetylcholinesterase (AChE), cholinesterases (ChE), aryleslerases, sterol esterases, insect juvenile hormone esterases, aixl others. The determination of A-esterases uses a protocol for the detection of PTEHs. The PTEH assay allows for the identification of two subclasses of esterases, the A-esterase (known as aiyldialkylphos-phatase) and ditsopropyl fluorophosphatase. Both these enzymes metabolize OP compounds. 

There are marked species differences in A-esterase activity. Birds have very low, often undetectable, levels of activity in plasma toward paraoxon, diazoxon, pirimi-phos-methyl oxon, and chlorpyrifos oxon (Brealey et al. 1980, Mackness et al. 1987, Walker et al. 1991 Figure 2.10). Mammals have much higher plasma A-esterase activities to all of these substrates. The toxicological implications of this are discussed in Chapter 10. Some species of insects have no measurable A-esterase activity, even in strains that have resistance to OPs (Mackness et al. 1982, Walker 1994). These include the peach potato aphid (Myzus persicae Devonshire 1991) and the... 

The organophosphorons insecticides dimethoate and diazinon are mnch more toxic to insects (e.g., housefly) than they are to the rat or other mammals. A major factor responsible for this is rapid detoxication of the active oxon forms of these insecticides by A-esterases of mammals. Insects in general appear to have no A-esterase activity or, at best, low A-esterase activity (some earlier stndies confnsed A-esterase activity with B-esterase activity) (Walker 1994b). Diazinon also shows marked selectivity between birds and mammals, which has been explained on the gronnds of rapid detoxication by A-esterase in mammals, an activity that is absent from the blood of most species of birds (see Section 23.23). The related OP insecticides pirimiphos methyl and pirimiphos ethyl show similar selectivity between birds and mammals. Pyrethroid insecticides are highly selective between insects and mammals, and this has been attributed to faster metabolic detoxication by mammals and greater sensitivity of target (Na+ channel) in insects. 

In addition to ester bonds with P (Section 10.2.1, Figures 10.1 and 10.2), some OPs have other ester bonds not involving P, which are readily broken by esteratic hydrolysis to bring about a loss of toxicity. Examples include the two carboxylester bonds of malathion, and the amido bond of dimethoate (Figure 10.2). The two carboxylester bonds of malathion can be cleaved by B-esterase attack, a conversion that provides the basis for the marked selectivity of this compound. Most insects lack an effective carboxylesterase, and for them malathion is highly toxic. Mammals and certain resistant insects, however, possess forms of carboxylesterase that rapidly hydrolyze these bonds, and are accordingly insensitive to malathion toxicity. 

Devonshire, A.L. (1991). Role of esterases in resistance of insects to insecticides. In Biochemical Society Transactions 19, 755-759. 

Metabolism of fenvalerate proceeds by way of oxidation and hydrolysis to produce metabolites considered pharmacologically inactive or inferior to the parent compound. Insects and fish are extremely susceptible to fenvalerate when compared to mammals and birds. Interspecies differences are associated with rates of metabolism, excretion, absorption, esterase activity, and neurosensitivity. 

Lord and Potter1 have claimed that it is important not to generalize the known anti-cholinesterase activity of organo-phosphorus insecticides in mammals to account for their action in insects. They could find no specific cholinesterase in two species of insect, but there was a general esterase inhibited by the insecticides. 

Carboxylesterases are well-represented in insects and are sometimes important in the development of resistance to insecticides. Thus, a well-characterized carboxylesterase E4 is responsible for resistance to organophosphorus insecticides in the aphid (Myzuspersicae) [107]. In the California Culex mosquito, the esterase B1 is 500-fold more abundant in organophosphate-resistant than in susceptible insects. The increase of esterase levels is the result of gene amplification, i.e., the resistant animals have an increased number of copies of the structural esterase gene [108],... 

Esterases of the Juvenile Hormone of Insects Many works have been dedicated to the inhibition of esterases of the juvenile hormone of insects. The purpose of these works is to control insect populations by ehminating their metamorphosis. Among the numerous trifluoromethyl ketones that have been synthesized, thioalkyl derivatives of trifluoroacetone have been shown to be the most active ones. Curiously, the corresponding alcohols are also excellent inhibitors. Trifluoromethyl ketones can also inhibit other insect esterases antenna esterases and esterases that are involved in the release of pheromones (Figure 7.33). The inhibition of these latter ones can also be interesting for insect control purposes. 

The mechanisms of resistance fall into two main categories. Many insects produce an increased level of detoxifying enzymes, such as esterases, that modify the insecticides to inactive metabolites very rapidly. Such a system is seen in aphids that are resistant to OP insecticides. In other cases it is the target site that is modified such that the insecticide (the enzyme inhibitor) no longer binds to the target and is, therefore, ineffective. This has recently been shown to occur in some aphids that are resistant to OP insecticides but the classical example is knockdown resistance (kdr) and super-kdr to pyrethroid insecticides shown by many insects but particularly house flies Musca domes tied). This resistance is thought to result from a modification of... 

Hydrolytic reactions. There are numerous different esterases responsible for the hydrolysis of esters and amides, and they occur in most species. However, the activity may vary considerably between species. For example, the insecticide malathion owes its selective toxicity to this difference. In mammals, the major route of metabolism is hydrolysis to the dicarboxylic acid, whereas in insects it is oxidation to malaoxon (Fig. 5.12). Malaoxon is a very potent cholinesterase inhibitor, and its insecticidal action is probably due to this property. The hydrolysis product has a low mammalian toxicity (see chap. 7). 

Another important problem is the development of insects resistant to insecticides. This often arises as a result of increased levels of carboxylesterases which hydrolyze both organophosphates and car-baryl.h/1 A mutation that changed a single active site glycine to aspartate in a carboxylesterase of a blowfly changed the esterase to an organophosphorus hydrolase which protected the fly against insecticides.)... 

One of the insect neurohormones, the activation hormone, controls the secretion of the corpora allata, paired glands that synthesize the juvenile hormone (Fig. 22-4) in insect larvae. While the structure of the juvenile hormone varies somewhat with species, it is usually a polyprenyl ester. A specific binding protein provides the hormone with protection from degrada-tive enzymes. However, in the tobacco homworm an esterase, able to hydrolyze the protein-bound juvenile hormone, is produced at the start of pupal differentiation.354 The exact mechanism of action of juvenile hormones has been difficult to determine. However, it affects polyamine synthesis.355 356... 

The hydrolysis of esters by esterases and of amides by amidases constitutes one of the most common enzymatic reactions of xenobiotics in humans and other animal species. Because both the number of enzymes involved in hydrolytic attack and the number of substrates for them is large, it is not surprising to observe interspecific differences in the disposition of xenobiotics due to variations in these enzymes. In mammals the presence of carboxylesterase that hydrolyzes malathion but is generally absent in insects explains the remarkable selectivity of this insecticide. As with esters, wide differences exist between species in the rates of hydrolysis of various amides in vivo. Fluoracetamide is less toxic to mice than to the American cockroach. This is explained by the faster release of the toxic fluoroacetate in insects as compared with mice. The insecticide dimethoate is susceptible to the attack of both esterases and amidases, yielding nontoxic products. In the rat and mouse, both reactions occur, whereas sheep liver contains only the amidases and that of guinea pig only the esterase. The relative rates of these degradative enzymes in insects are very low as compared with those of mammals, however, and this correlates well with the high selectivity of dimethoate. 

Blood and various organs of humans and other animals contain esterases capable of acetylsalicylic acid hydrolysis. A comparative study has shown that the liver is the most active tissue in all animal species studied except for the guinea pig, in which the kidney is more than twice as active as the liver. Human liver is least active the enzyme in guinea pig liver is the most active. The relatively low toxicity of some of the new synthetic pyrethroid insecticides appears to be related to the ability of mammals to hydrolyze their carboxyester linkages. Thus mouse liver microsomes catalyzing (+)-/runs-resin e 111ri n hydrolysis are more than 30-fold more active than insect microsomal preparations. The relative rates of hydrolysis of this substrate in enzyme preparations from various species are mouse > > milkweed bug > > cockroach > > cabbage looper > housefly. 

Ishida Y. and Leal S. (2002) Cloning of putative odorant-degrading enzyme and integumental esterase cDNAs from the wild silkmoth, Antheraea polyphemus. Insect Biochem. Mol. Biol. 32, 1775-1780. 

Since this manuscript was submitted, the ApolSE has reportedly been cloned, using PCR primers designed to conserved regions of known insect esterase enzymes (Ishida and Leal, 2002), indicating that ApolSE is indeed a member of... 

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