Method for improving polyphenol oxidase activity in apples by using cetyltrimethylammonium bromide and dodecyltrimethylammonium bromide

A technology of cetyltrimethylammonium bromide and dodecyltrimethylammonium bromide, which is applied in the field of enzyme catalyst preparation in bioengineering, can solve the problems of low PPO activity, reduced content of phenolic compounds, and enzyme Activity has an impact, etc.

Active Publication Date: 2019-06-21
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, among which apples are rich in content. my country's apple planting area and output account for more than 40% of the world's total, but the activity of PPO extracted from apples is low

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  • Method for improving polyphenol oxidase activity in apples by using cetyltrimethylammonium bromide and dodecyltrimethylammonium bromide
  • Method for improving polyphenol oxidase activity in apples by using cetyltrimethylammonium bromide and dodecyltrimethylammonium bromide
  • Method for improving polyphenol oxidase activity in apples by using cetyltrimethylammonium bromide and dodecyltrimethylammonium 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 12 h).

[0023] 2. Extraction of crude enzyme solution: Weigh 58.82 g of treated apples, place them in 100 mL of phosphate buffered saline solution (0.2 M, pH6.5), beat with a mixer, and then filter with four layers of gauze. The obtained crude filtrate was centrifuged at 10,000 rpm for 10 min at 4°C, and the supernatant, ie the PPO crude enzyme solution, was taken for later use.

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

[0025] 3. Preparation of surfactant solution:

[0026] Prepare 25 mM CTAB and 60 mM DTAB 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 ultraviolet spectropho...

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Abstract

The invention discloses a method for improving the activity of polyphenol oxidase in apples by using cetyltrimethylammonium bromide and dodecyltrimethylammonium bromide, belonging to the technical field of enzyme catalyst preparation in bioengineering. The present invention includes the extraction of polyphenol oxidase (PPO) crude enzyme liquid, characterized in that: Under certain conditions, the catalytic reaction is carried out with polyphenol oxidase as a catalyst. The invention utilizes cetyltrimethylammonium bromide or dodecyltrimethylammonium bromide to improve the activity of PPO in apples.

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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02
CPCC12N9/0059C12Y110/03001
Inventor 郭霞张礼飞
Owner YANGZHOU UNIV
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