Purification and characterization of an acetylcholinesterase from the infective juveniles of Heterorhabditis bacteriophora

Mohamed, M. I.; Abdel-Gawad, A. S.; Ghazy, Abdel-Hady M.

doi:10.1016/j.cbpc.2007.04.001

Cited by 9 publications

(4 citation statements)

References 40 publications

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“…ATC was shown to have the greatest ratio of catalytic efficiency in a series of calculations, thus proving that a partial purified enzyme sample collected from brain extract of P. reticulata predominantly contains AChE. This observation is in agreement with the data obtained in several studies [4,22,[25][26][27][28][29][30][31][32]. The result also showed that the ChE in brain extract prefers to hydrolyse ATC, followed by PTC and BTC.…”

Section: Substrate Specificity Profilesupporting

confidence: 89%

Partial Purification and Characterization of the Acetylcholinesterase from Poecilia reticulata (Peters, 1859)

Hazuki,

Yasid,

Shukor

2023

J Environ Bioremed Toxicol

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Poecilia reticulata (P. reticulata) is a species of tropical fish in the family Poeciliidae that is found all over the world and is a favorite in freshwater aquariums. The Malay name for this fish is "Ikan Gapi," although the English term "Guppy" has become more common. The Million Fish, or Rainbow Fish, as it's known in other parts of the world. The goals of this research were to char-acterize the acetylcholinesterase that will be partially purified from P. reticulata brain extract using ammonium sulphate fractionation followed by an affinity chromatography method. The result shows that ammonium sulphate fractionation gave better purification fold over the procainamide affinity chromatographic method with fold of purification of 3.38 compared to 0.64 for the latter. Polyacrylamide gel electrophoresis (SDS-PAGE) shows a dramatic reduction of protein bands compared to crude extract. A pH of 9 in a Tris-HCl buffer system and a temperature of 15 °C were found to be ideal for the activity of the enzyme. Based on its high Vmax and low Km with acetylthiocholine iodide (ATC) as a substrate compared to butyryl thiocholine iodide (BTC) and propionyl thiocholine iodide (PTC), the substrate specificity profile identified acetylcholinesterase (AChE) as the major enzyme present in the purified enzyme. The molecular weights of the en-zyme following affinity chromatographic purification are about 92 kDa.

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Section: Substrate Specificity Profilesupporting

confidence: 89%

Partial Purification and Characterization of the Acetylcholinesterase from Poecilia reticulata (Peters, 1859)

Hazuki,

Yasid,

Shukor

2023

J Environ Bioremed Toxicol

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“…(). A sample of L 4 gut extract (20 μg of protein) was submitted to SDS‐PAGE (Mohamed et al., ) using a 12.5% (w/v) gel at 4°C. After electrophoresis, the gel was immersed in 2.5% Triton X‐100 in 0.1 M Tris‐HCl pH 8.0 to remove SDS and incubated (30 min, 4°C) with 3% casein (w/v) in 0.1 M Tris‐HCl pH 8.0.…”

Section: Methodsmentioning

confidence: 99%

“…Zymography for AChE was performed according to Mohamed et al. (). Whole L 4 extract (100 μg of protein) was submitted to SDS‐PAGE on 6–10% (w/v) gradient gel.…”

Section: Methodsmentioning

confidence: 99%

EFFECT OF Moringa oleifera FLOWER EXTRACT ON LARVAL TRYPSIN AND ACETHYLCHOLINESTERASE ACTIVITIES IN Aedes aegypti

Pontual

Napoleão

Assis

et al. 2012

Arch Insect Biochem Physiol

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Aedes aegypti control is crucial to reducing dengue fever. Aedes aegypti larvae have developed resistance to organophosporous insecticides and the use of natural larvicides may help manage larval resistance by increasing elements in insecticide rotation programs. Here, we report on larvicidal activity of Moringa oleifera flower extract against A. aegypti L(1), L(2), L(3), and L(4) as well as the effect of flower extract on gut trypsin and whole-larval acetylcholinesterase from L(4.) In addition, the heated flower extract was investigated for larvicidal activity against L(4) and effect on larval gut trypsin. Moringa oleifera flower extract contains a proteinaceous trypsin inhibitor (M. oleifera flower trypsin inhibitor, MoFTI), triterpene (β-amyrin), sterol (β-sitosterol) as well as flavonoids (kaempferol and quercetin). Larvicidal activity was detected against L(2), L(3), and L(4) (LC(50) of 1.72%, 1.67%, and 0.92%, respectively). Flower extract inhibited L(4) gut trypsin (MoFTI K(i) = 0.6 nM) and did not affect acetylcholinesterase activity. In vivo assay showed that gut trypsin activity from L(4) treated with M. oleifera flower extract decreased over time (0-1,440 min) and was strongly inhibited (98.6%) after 310 min incubation; acetylcholinesterase activity was not affected. Thermal treatment resulted in a loss of trypsin inhibitor and larvicidal activities, supporting the hypothesis that flower extract contains a proteinaceous trypsin inhibitor that may be responsible for the deleterious effects on larval mortality.

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“…The further purification was carried out using size exclusion chromatography on a Sephacryl S-100 (Figure 1A), column equilibrated with Tris-HCl 20mM in Triton x100 (0,1%), NaCl 0.25M, pH 8.0. AChE was eluted using the same buffer and the chromatographic step decreased the specific activity to 0.194 U/ mg [8,9].…”

Section: Partial Purification Of Prochilodus Brevis Achementioning

confidence: 99%

Acetylcholinesterase from Curimatã Fish Brain (Prochilodus Brevis) as Potential Biocatalyst for Voltammetric Biosensor Construction

Abreu

2018

BJSTR

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Background:The AChE (acetylcholinesterase) is a serine hydrolase responsible for terminating neurotransmission by hydrolyzing the acetylcholine released on synaptic cleft. Studies of AChE as a target of pesticide toxicity have yielded several practical outcomes and are the basis for constructing biosensors. These devices are primarily designed to determine and quantify the inhibition of AChE by toxic chemicals.Objective: to construct a biosensor based on acetylcholinesterase from the brain of the Prochilodus brevis fish, and to use the same as biomarker of agrochemicals that inhibit the enzyme.Methods: Acetylcholinesterase was isolated from curimatã fish brain (prochilodus brevis) and partially purified using amonium sulfate precipitation followed by size-exclusion chromatography (ChE1). AChE from curimatã fish brain was directly immobilizes on the surface of glassy carbon electrode modified with multi-walled carbon nanotubes.Results: Acetylcholinesterase was characterized as having a specific activity of 0.194U/mg. The optimum activity was found at pH 8.5, phosphate buffer 0.7µM, 28ºC and exhibited a thermostability at 37°C. The glassy carbon modified electrode exhibits excellent electrocatalytic activity to the increase of thiocholine, with a linear response in the 0.05 mM to 0.85 mM concentration range, with a 73µM limit of detection and with a 240µM limit of quantification. Conclusion:The AChE from curimatã fish brain allowed the modification of the glassy carbon electrode providing a potential sensor detection system that can be used for determination of thiocholine (pesticides) in real samples of environmental importance.

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Purification and characterization of an acetylcholinesterase from the infective juveniles of Heterorhabditis bacteriophora

Cited by 9 publications

References 40 publications

Partial Purification and Characterization of the Acetylcholinesterase from Poecilia reticulata (Peters, 1859)

Partial Purification and Characterization of the Acetylcholinesterase from Poecilia reticulata (Peters, 1859)

EFFECT OF Moringa oleifera FLOWER EXTRACT ON LARVAL TRYPSIN AND ACETHYLCHOLINESTERASE ACTIVITIES IN Aedes aegypti

Acetylcholinesterase from Curimatã Fish Brain (Prochilodus Brevis) as Potential Biocatalyst for Voltammetric Biosensor Construction

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