CN102477413A - Separation technology for polyphenol oxidase of tobacco - Google Patents
Separation technology for polyphenol oxidase of tobacco Download PDFInfo
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- CN102477413A CN102477413A CN2010105718176A CN201010571817A CN102477413A CN 102477413 A CN102477413 A CN 102477413A CN 2010105718176 A CN2010105718176 A CN 2010105718176A CN 201010571817 A CN201010571817 A CN 201010571817A CN 102477413 A CN102477413 A CN 102477413A
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- tobacco
- polyphenoloxidase
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Abstract
The invention relates to tobacco, in particular to a separation technology for polyphenol oxidase of the tobacco, which belongs to the technical field of chemical industry. In the technology, a finished product is prepared by adopting the following steps: chopping, homogenating and freezing fresh tobacco, centrifugally extracting supernate through crude enzyme extraction and ultrasonic treatment to perform chromatographic separation and condensing. The separation technology has a simple process and a reasonable flow. The obtained polyphenol oxidase can be widely used for medicines, foodstuffs and relative Industry.
Description
Technical field
The present invention relates to one and grow tobacco, be specifically related to one grow tobacco polyphenoloxidase stripping technique, belong to chemical technology field.
Background technology
Polyphenoloxidase is the one type of cuproprotein material that extensively exists in the plant, is oxidized to quinone through the molecular oxygen catalysis aldehydes matter, and polymerization and the brownization of quinone takes place subsequently.Several kinds of different polyphenoloxidase are all arranged in different plants.Owing to the main effect of polyphenoloxidase is the spoiled and loss of nutrient substances that the enzyme browning reaction causes fruit, vegetables.Polyphenoloxidase is not only served as vital role in the developmental process of tobacco, and affects the color and the fragrance of tobacco leaf.Because polyphenoloxidase potential application background reaches the freshening effect to food, in recent years, for from tobacco, extracting polyphenoloxidase, is in the development stage.
Summary of the invention
The objective of the invention is for overcoming the weak point of above-mentioned prior art; Provide one grow tobacco polyphenoloxidase stripping technique; This technology adopts new fresh tobacco leaf chopping, homogenate, freezing, through the extraction of crude enzyme liquid, through ultrasonication; Centrifugal extraction supernatant is carried out chromatographic separation, gets finished product after concentrating.
The present invention realizes with following technical scheme: one grow tobacco polyphenoloxidase stripping technique, it is characterized in that: this technology comprises the steps:
(1) extraction of crude enzyme liquid, with new fresh tobacco leaf chopping, homogenate under 4 ℃ of conditions adds cold acetone and water, under-18 ℃ of conditions, preserves 24h;
(2) with 4 layers of filtered through gauze of step (1) warp,, freezing subsequent use behind the dry 2h under the room temperature with 150ml cold acetone washing 3 times;
(3) phosphoric acid buffer of adding 200ml in above-mentioned acetone dry powder, pH value is 6.5, ultrasonication 1h; Centrifugal; The ammonium sulfate that adds 30% volumetric molar concentration in the supernatant, centrifugal behind the stirring 4h under 4 ℃ of conditions, get supernatant; Get the ammonium sulfate of 80% volumetric molar concentration, centrifugal behind the stirring 24h under 4 ℃ of conditions;
(4) above-mentioned solution is carried out chromatographic separation, wherein polyphenoloxidase is preserved subsequent use under-18 ℃ of conditions after the dialysis tubing desalination and concentration.
Advantage of the present invention is: this technology adopts new fresh tobacco leaf chopping, homogenate, freezing, and through the extraction of crude enzyme liquid, through ultrasonication, centrifugal extraction supernatant is carried out chromatographic separation, gets finished product after concentrating.This stripping technique, technology is simple, and flow process is reasonable, and the gained polyphenoloxidase can be widely used in medicine, food and association area.
Embodiment
Embodiment,
One grow tobacco polyphenoloxidase stripping technique, it is characterized in that: this technology comprises the steps:
(1) extraction of crude enzyme liquid, with new fresh tobacco leaf chopping, homogenate under 4 ℃ of conditions adds cold acetone and water, under-18 ℃ of conditions, preserves 24h;
(2) with 4 layers of filtered through gauze of step (1) warp,, freezing subsequent use behind the dry 2h under the room temperature with 150ml cold acetone washing 3 times;
(3) phosphoric acid buffer of adding 200ml in above-mentioned acetone dry powder, pH value is 6.5, ultrasonication 1h; Centrifugal; The ammonium sulfate that adds 30% volumetric molar concentration in the supernatant, centrifugal behind the stirring 4h under 4 ℃ of conditions, get supernatant; Get the ammonium sulfate of 80% volumetric molar concentration, centrifugal behind the stirring 24h under 4 ℃ of conditions;
(4) above-mentioned solution is carried out chromatographic separation, wherein polyphenoloxidase is preserved subsequent use under-18 ℃ of conditions after the dialysis tubing desalination and concentration.
Claims (1)
- One grow tobacco polyphenoloxidase stripping technique, it is characterized in that: this technology comprises the steps:(1) extraction of crude enzyme liquid, with new fresh tobacco leaf chopping, homogenate under 4 ℃ of conditions adds cold acetone and water, under-18 ℃ of conditions, preserves 24h;(2) with 4 layers of filtered through gauze of step (1) warp,, freezing subsequent use behind the dry 2h under the room temperature with 150ml cold acetone washing 3 times;(3) phosphoric acid buffer of adding 200ml in above-mentioned acetone dry powder, pH value is 6.5, ultrasonication 1h; Centrifugal; The ammonium sulfate that adds 30% volumetric molar concentration in the supernatant, centrifugal behind the stirring 4h under 4 ℃ of conditions, get supernatant; Get the ammonium sulfate of 80% volumetric molar concentration, centrifugal behind the stirring 24h under 4 ℃ of conditions;(4) above-mentioned solution is carried out chromatographic separation, wherein polyphenoloxidase is preserved subsequent use under-18 ℃ of conditions after the dialysis tubing desalination and concentration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105718176A CN102477413A (en) | 2010-11-25 | 2010-11-25 | Separation technology for polyphenol oxidase of tobacco |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105718176A CN102477413A (en) | 2010-11-25 | 2010-11-25 | Separation technology for polyphenol oxidase of tobacco |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102477413A true CN102477413A (en) | 2012-05-30 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010105718176A Pending CN102477413A (en) | 2010-11-25 | 2010-11-25 | Separation technology for polyphenol oxidase of tobacco |
Country Status (1)
| Country | Link |
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| CN (1) | CN102477413A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105158180A (en) * | 2015-07-07 | 2015-12-16 | 莆田学院 | Method for screening tyrosinase inhibitor |
| CN106754771A (en) * | 2016-12-13 | 2017-05-31 | 罗志聪 | The extracting method of polyphenol oxidase in a kind of lychee exocarp |
| CN113017140A (en) * | 2021-03-12 | 2021-06-25 | 云南瑞升烟草技术（集团）有限公司 | Method for preparing endogenous total enzyme by using fresh tobacco and application of endogenous total enzyme |
-
2010
- 2010-11-25 CN CN2010105718176A patent/CN102477413A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105158180A (en) * | 2015-07-07 | 2015-12-16 | 莆田学院 | Method for screening tyrosinase inhibitor |
| CN106754771A (en) * | 2016-12-13 | 2017-05-31 | 罗志聪 | The extracting method of polyphenol oxidase in a kind of lychee exocarp |
| CN113017140A (en) * | 2021-03-12 | 2021-06-25 | 云南瑞升烟草技术（集团）有限公司 | Method for preparing endogenous total enzyme by using fresh tobacco and application of endogenous total enzyme |
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Legal Events
| Date | Code | Title | Description |
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| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20120530 |