The anti-cholinesterase
activity of the obtained products was accomplished according to the
described procedure.36 (link) Electric eel AChE
(type-VI-S), potassium phosphate buffer (pH 8.0), equine BChE, butyryl
thiocholine iodide, 5,5-dithio-bis-nitrobenzoic acid
(DTNB), galantamine from Lycoris sp.,
and acetyl thiocholine iodide, were used in the present study.
Spectrophotometric Ellman’s technique was used to carry out
the AChE and BChE assays using butyryl thiocholine iodide and acetyl
thiocholine iodide as substrates. Two mixtures having 5 μL of
AChE (0.03 μg/mL) and 5 μL of BChE (0.01 μg/mL)
particularly and each one added with 205 μL of samples (125–1000
μg/mL) and DTNB (5 μL) were incubated for 15 min at 30
°C. After 15 min, 5 μL of substrates was added to the reaction
mixture, and the reaction was detected for 4 min at 412 nm by using
a UV–visible spectrophotometer. The presence of yellow color
in the mixture showed the creation of the 5-thio-2-nitrobenzoate anion
by reaction of thiocholines and DTNB. In the absence of samples and
enzymes, nonenzymatic hydrolysis of substrate was also examined. The
reaction mixture was considered as a control in the absence of samples.
Percent inhibition and enzyme activities were calculated through the
following formulas where V is the rate of reaction
in the presence of an inhibitor and Vmax is the rate of reaction without the inhibitor.
activity of the obtained products was accomplished according to the
described procedure.36 (link) Electric eel AChE
(type-VI-S), potassium phosphate buffer (pH 8.0), equine BChE, butyryl
thiocholine iodide, 5,5-dithio-bis-nitrobenzoic acid
(DTNB), galantamine from Lycoris sp.,
and acetyl thiocholine iodide, were used in the present study.
Spectrophotometric Ellman’s technique was used to carry out
the AChE and BChE assays using butyryl thiocholine iodide and acetyl
thiocholine iodide as substrates. Two mixtures having 5 μL of
AChE (0.03 μg/mL) and 5 μL of BChE (0.01 μg/mL)
particularly and each one added with 205 μL of samples (125–1000
μg/mL) and DTNB (5 μL) were incubated for 15 min at 30
°C. After 15 min, 5 μL of substrates was added to the reaction
mixture, and the reaction was detected for 4 min at 412 nm by using
a UV–visible spectrophotometer. The presence of yellow color
in the mixture showed the creation of the 5-thio-2-nitrobenzoate anion
by reaction of thiocholines and DTNB. In the absence of samples and
enzymes, nonenzymatic hydrolysis of substrate was also examined. The
reaction mixture was considered as a control in the absence of samples.
Percent inhibition and enzyme activities were calculated through the
following formulas where V is the rate of reaction
in the presence of an inhibitor and Vmax is the rate of reaction without the inhibitor.
