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. 2014:2014:571094.
doi: 10.1155/2014/571094. Epub 2014 Oct 27.

The assessment of cholinesterase from the liver of Puntius javanicus as detection of metal ions

Mohd Khalizan Sabullah  1 , Mohd Rosni Sulaiman  2 , Mohd Yunus Abd Shukor  3 , Mohd Arif Syed  3 , Nor Aripin Shamaan  4 , Ariff Khalid  5 , Siti Aqlima Ahmad  3
Affiliations

The assessment of cholinesterase from the liver of Puntius javanicus as detection of metal ions

Mohd Khalizan Sabullah et al. ScientificWorldJournal. 2014.

Abstract

Crude extract of ChE from the liver of Puntius javanicus was purified using procainamide-sepharyl 6B. S-Butyrylthiocholine iodide (BTC) was selected as the specific synthetic substrate for this assay with the highest maximal velocity and lowest biomolecular constant at 53.49 µmole/min/mg and 0.23 mM, respectively, with catalytic efficiency ratio of 0.23. The optimum parameter was obtained at pH 7.5 and optimal temperature in the range of 25 to 30°C. The effect of different storage condition was assessed where ChE activity was significantly decreased after 9 days of storage at room temperature. However, ChE activity showed no significant difference when stored at 4.0, 0, and -25°C for 15 days. Screening of heavy metals shows that chromium, copper, and mercury strongly inhibited P. javanicus ChE by lowering the activity below 50%, while several pairwise combination of metal ions exhibited synergistic inhibiting effects on the enzyme which is greater than single exposure especially chromium, copper, and mercury. The results showed that P. javanicus ChE has the potential to be used as a biosensor for the detection of metal ions.

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Figures

Figure 1
Figure 1
Profile of procainamide-based affinity chromatography purification on ChE from P. javanicus liver. Error bars represent mean ± standard error (n = 3).
Figure 2
Figure 2
Three synthetic substrates, namely, acetylthiocholine iodide (ATC), butyrylthiocholine iodide (BTC), and propionylthiocholine iodide (PTC), with different concentrations ranging from 0 to 2.5 mM incubated with ChE to prove its specificity. Error bars represent mean ± standard error (n = 3).
Figure 3
Figure 3
Optimisation of pH for P. javanicus ChE. Error bars represent mean ± standard error (n = 3).
Figure 4
Figure 4
Optimisation of temperature for ChE from P. javanicus. Error bars represent mean ± standard error (n = 3).
Figure 5
Figure 5
Storage condition of ChE at different temperature: (black) room temperature, (dark grey) 4°C, (light grey) 0°C, and (white) −25°C. Error bars represent mean ± standard error (n = 3).
Figure 6
Figure 6
Percentage of enzyme activity after inhibition by heavy metals at 5 mg/L final concentration. Error bars represent mean ± standard error (n = 3).
Figure 7
Figure 7
Synergistic reaction of P. javanicus ChE with the pairwise metal ion. Error bars represent mean ± standard error (n = 3).
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