Application of 4-sulfonated calix [n] arene/CTAB complex system in improving activity of polyphenol oxidase
A technology of polyphenol oxidase and sulfonated cup, which is applied in the field of biological enzymes, can solve the problems of polyphenol oxidase enzyme activity reduction and inactivation, and achieve the effect of improving enzyme activity
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[0023] (1) Preparation of crude enzyme solution
[0024] Crude pineapple extract: Peel the pineapple and cut into pieces, weigh 24.3158g, add 24mL of phosphate buffer solution (40mM, pH6.0) containing disodium ethylenediaminetetraacetic acid (EDTA·2Na), squeeze it with a juicer Juice for 3-5 minutes, then filter with gauze, centrifuge the filtrate in a high-speed refrigerated centrifuge with a rotation speed of 10,000 rpm and a temperature of 4°C for 25 minutes, and store the separated supernatant at -20°C.
[0025] Crude Apple Extract: Wash the apples in water, drain, and freeze at -20°C for 12 hours. Peel 58.82 g of frozen apples, add 100 mL of phosphate buffer solution (0.2M pH=6.5, containing 0.25% (v / v) TritonX-100 and 2% (w / w) polyvinylpyrrolidone) and mix for 1 min. Then filter with gauze, centrifuge the filtrate in a high-speed refrigerated centrifuge with a rotation speed of 10,000 rpm and a temperature of 4°C for 10 minutes, and store the separated supernatant at -2...
Embodiment 1
[0037] Effect of SC[4] / CTAB compound system on polyphenol oxidase activity in pineapple crude extract:
[0038] The experiment was divided into two groups: group 1 mixed a certain amount of SC[4] mother liquor and pineapple crude extract, group 2 mixed a certain amount of CTAB mother liquor, SC[4] mother liquor and pineapple crude extract, and equilibrated at 30°C for 1h . The obtained mixed solution is added into the catechol solution to carry out the catalytic reaction, and after the reaction is completed, the reaction solution is detected at 420 nm for the change of the absorbance value with time. In the total reaction system, the concentrations of SC[4] in group 1 were 0, 0.02mg / mL, 0.06mg / mL, 0.08mg / mL, 0.1mg / mL, and the concentration of catechol was 150mM. In group 2, the concentration of CTAB was 0.1mg / mL, the concentration of SC[4] was 0, 0.02mg / mL, 0.06mg / mL, 0.08mg / mL, 0.1mg / mL, and the concentration of catechol was 150mM . The absorbance value (A) at the 420nm pl...
Embodiment 2
[0040] Effect of SC[6] / CTAB compound system on polyphenol oxidase activity in pineapple crude extract:
[0041]The experiment was divided into two groups: group 1 mixed a certain amount of SC[6] mother liquor and pineapple crude extract, group 2 mixed a certain amount of CTAB mother liquor, SC[6] mother liquor and pineapple crude extract, and equilibrated at 30°C 1h. The obtained mixed solution is added into the catechol solution to carry out the catalytic reaction, and after the reaction is completed, the reaction solution is detected at 420 nm for the change of the absorbance value with time. In the total reaction system, in group 1, the concentrations of SC[6] were 0, 0.02mg / mL, 0.06mg / mL, 0.08mg / mL, 0.1mg / mL, and the concentration of catechol was 150mM. In group 2, the concentration of CTAB was 0.1mg / mL, the concentration of SC[6] was 0, 0.02mg / mL, 0.06mg / mL, 0.08mg / mL, 0.1mg / mL, and the concentration of catechol was 150mM . The absorbance value (A) at the 420nm place i...
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