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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

Pending Publication Date: 2019-10-11
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is widely used in many fields such as food, chemical industry, medicine, etc., but because some additives are often used in the application process, the enzyme activity of polyphenol oxidase is reduced or even inactivated

Method used

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  • Application of 4-sulfonated calix [n] arene/CTAB complex system in improving activity of polyphenol oxidase
  • Application of 4-sulfonated calix [n] arene/CTAB complex system in improving activity of polyphenol oxidase
  • Application of 4-sulfonated calix [n] arene/CTAB complex system in improving activity of polyphenol oxidase

Examples

Experimental program
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Effect test

preparation example Construction

[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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Abstract

The invention discloses application of a 4-sulfonated calix [n] arene / CTAB complex system in improving activity of polyphenol oxidase in pineapples. By complexing 4-sulfonated calix [n] arene inhibiting enzymatic activity and a surfactant CTAB improving the enzymatic activity, a super-amphiphilic molecule is formed to react with the polyphenol oxidase in a pineapple extracting solution. The regulation of the activity of the polyphenol oxidase in the pineapples is achieved through the 4-sulfonated calix [n] arene / CTAB complex system for the first time, complexing of the 4-sulfonated calix [n] arene / CTAB reduces the inhibiting effect of the 4-sulfonated calix [n] arene on the activity of the polyphenol oxidase in the pineapples, and under optimal conditions, the enzymatic activity of the polyphenol oxidase in the pineapples can be significantly increased to 419%.

Description

technical field [0001] The invention belongs to the technical field of biological enzymes, and relates to the application of a compound system of 4-sulfonated calix[n]arene / hexadecyltrimethylammonium bromide in improving the activity of polyphenol oxidase, in particular to 4-sulfonated calix [n]Application of aromatic hydrocarbon / cetyltrimethylammonium bromide compound system in improving the activity of polyphenol oxidase in pineapple. Background technique [0002] There are many different kinds of proteases in many plants, among which polyphenol oxidase (PPO) is a kind of metalloprotease widely distributed in nature, and widely exists in plants such as apples, pineapples, and mushrooms. It is widely used in food, chemical industry, medicine and many other fields, but because some additives are often used in the application process, the enzyme activity of polyphenol oxidase is reduced or even inactivated. How to maintain the activity of polyphenol oxidase in the presence o...

Claims

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Application Information

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IPC IPC(8): C12N9/02
CPCC12N9/0059C12Y110/03001
Inventor 郭霞朱嘉昕陈露
Owner YANGZHOU UNIV
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