Method for improving activity of polyphenol oxidase in apples by virtue of hexadecyl trimethyl ammonium bromide and dodecyl trimethyl ammonium bromide

A technology of dodecyltrimethylammonium bromide and hexadecyltrimethylammonium bromide, which is applied in the field of enzyme catalyst preparation in bioengineering, can solve the problems of enzyme activity, low PPO activity, phenolic Compound content reduction and other issues

Active Publication Date: 2016-07-06
YANGZHOU UNIV
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Problems solved by technology

However, after purification, the content of phenolic compounds is reduced and polyphenol oxidase (PPO) is in a bound state, which may affect the enzyme activity.
[0005] PPO exists in a large amount in plants

Method used

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  • Method for improving activity of polyphenol oxidase in apples by virtue of hexadecyl trimethyl ammonium bromide and dodecyl trimethyl ammonium bromide
  • Method for improving activity of polyphenol oxidase in apples by virtue of hexadecyl trimethyl ammonium bromide and dodecyl trimethyl ammonium bromide
  • Method for improving activity of polyphenol oxidase in apples by virtue of hexadecyl trimethyl ammonium bromide and dodecyl trimethyl ammonium bromide

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Experimental program
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Embodiment Construction

[0021] 1. Preparation of polyphenol oxidase crude enzyme solution:

[0022] 1. Pretreatment: Wash the apples in distilled water, dry them, and store them in a freezer at -20°C (more than 12h).

[0023] 2, the extraction of crude enzyme liquid: take by weighing 58.82g processed apples, place in 100mL phosphate buffered saline solution (0.2M, pH6.5), make a slurry with a mixer, then squeeze and filter with four layers of gauze to obtain The crude filtrate was centrifuged at 10,000 rpm for 10 min at 4°C, and the supernatant, that is, the PPO crude enzyme solution, was taken for later use.

[0024] 2. Preparation of catechol solution: prepare 300mM catechol solution in phosphate buffer solution (0.1M, pH7).

[0025] 3. Preparation of surfactant solution:

[0026] Prepare 25 mMCTAB and 60 mMDTAB solutions in deionized water, respectively.

[0027] 4. Carry out enzyme-catalyzed reaction test with catechol as substrate, and measure the activity of PPO enzyme respectively with ultr...

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Abstract

The invention relates to a method for improving the activity of polyphenol oxidase in apples by virtue of hexadecyl trimethyl ammonium bromide and dodecyl trimethyl ammonium bromide, belonging to the technical field of preparation of enzyme catalysts in bioengineering. The method comprises the step of extraction of polyphenol oxidase (PPO) crude enzyme liquid. The method is characterized in that catalytic reaction is carried out by taking polyphenol oxidase as a catalyst in the presence of hexadecyl trimethyl ammonium bromide (CTAB) or dodecyl trimethyl ammonium bromide (DTAB). According to the method, the activity of polyphenol oxidase in the apples is improved by virtue of hexadecyl trimethyl ammonium bromide or dodecyl trimethyl ammonium bromide.

Description

technical field [0001] The invention belongs to the technical field of preparation of enzyme catalysts in bioengineering. Background technique [0002] In recent years, researchers have explored the mechanisms of various enzymes through enzyme-catalyzed reactions, which can not only reveal the mystery of the biocatalysis process, but also lay a scientific foundation for human beings to use some of these principles to research and develop new and efficient catalysts. [0003] Polyphenol oxidase is a kind of metalloprotease widely distributed in nature. It is ubiquitous in plants and can oxidize phenols or polyphenols to corresponding quinones. It is widely used in food, medicine and environmental protection industries. Researching its activity can better solve problems such as food storage, plant resistance to pathogens and viruses, and biodegradation of industrial wastewater containing phenolic organic substances. [0004] At present, most researchers study the purified pol...

Claims

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Application Information

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IPC IPC(8): C12N9/02
CPCC12N9/0059C12Y110/03001
Inventor 郭霞张礼飞
Owner YANGZHOU UNIV
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