CN101701234A - Method for extracting ellagic acid in pomegranate peel residue by using biological enzyme - Google Patents
Method for extracting ellagic acid in pomegranate peel residue by using biological enzyme Download PDFInfo
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- CN101701234A CN101701234A CN200910236945A CN200910236945A CN101701234A CN 101701234 A CN101701234 A CN 101701234A CN 200910236945 A CN200910236945 A CN 200910236945A CN 200910236945 A CN200910236945 A CN 200910236945A CN 101701234 A CN101701234 A CN 101701234A
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- ellagic acid
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- ellagitannins
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Abstract
The invention discloses a method for extracting ellagic acid in pomegranate peel residue by using biological enzyme, belonging to the fields of food, medicine and chemical engineering. The method takes the pomegranate peel residue as raw material and water as solvent, and can be used for preparing the ellagic acid by using the biological enzyme. Biological compound enzyme of degradable ellagitannin can be used for carrying out enzymolysis reaction on the ellagitannin, and alkaline liquor is used for stopping the enzymolysis reaction; simultaneously, an alkaline water system can be formed for extracting the ellagic acid, and crude product of the ellagic acid can be obtained; and the extraction rate of the ellagic acid reaches 12.69-24% based on the content of the ellagitannin obtained in the step (1). The method has simple and continuous technique, environment-friendly manufacturing process, environmental protection and less discharge of the three wastes, and is suitable for large-scale industrialized production.
Description
Technical field
The invention belongs to food, medicine, chemical field, be specifically related to a kind of method of extracting ellagic acid in the pomegranate peel residue with biological enzyme.
Background technology
Ellagic acid (2,3,7,8-tet rahydroxy benzopyrano[5,4,3cde] benzopyran-5, be the dimerization derivative of gallic acid 10-dione), be a kind of natural polyphenol material that extensively is present in the plant tissues such as various mushy fruits, nut.At nature, it can not only exist with the free form, and more is that form with the Ellagitannins of facile hydrolysis exists.The molecular formula of ellagic acid is C
14H
6O
8, be slightly soluble in water, alcohol, be dissolved in alkali, pyridine, be insoluble to ether.Pure ellagic acid is a kind of yellow needle-like crystal, and fusing point (pyridine) is greater than 360 ℃.
The poly-hydroxy structures shape of ellagic acid it have multiple physiologically active.Studies show that ellagic acid has effects such as good inhibition tumour, anti-oxidant, blood coagulation, inhibition bacterium and virus, calm, step-down.Thereby it is widely used in fields such as food, medicine, chemical industry.But be subjected to the restriction of ellagic acid physico-chemical property, its extraction, purifying never have good technical and break through, and have limited the production and the application of the ellagic acid of plant origin.
Preparation method according to the bibliographical information ellagic acid mainly contains following several:
A kind of is to utilize chemical solvents to extract from plant tissue, can utilize raw material production purity such as pomegranate, strawberry than high product by this method, it is that raw material can be produced content and is about 5% natural ellagic acid extract with strawberry, pomegranate that four or five tame factories are arranged at present in the world approximately.But owing to be subjected to the restriction of ellagic acid content in the raw material, can't produce on a large scale, and chemical solvents can cause detrimentally affect to environment, historical facts or anecdotes border using value is little.
A kind of is to obtain ellagic acid by the acidolysis Ellagitannins.Ellagitannins is heated under acidic conditions and glucose response can obtain ellagic acid, this method productive rate is higher relatively, but the acidolysis process need at high temperature carries out, and acid-resistant system costs an arm and a leg, and does not launch suitability for industrialized production as yet.
Microbial degradation method is to utilize the microorganism can produce tannase mainly to have aspergillus niger, wooden aspergillus, Penicillium chrysogenum, wooden mould, whiterot fungi and candiyeast etc. to act on ester bond in the tannin molecule, and Ellagitannins is degraded to ellagic acid.But ellagic acid is water insoluble, and fermenting process also has a large amount of mycelium to produce, and this method long reaction time, for follow-up separation and purification has brought very big difficulty, is difficult to obtain highly purified ellagic acid.
In addition, can also obtain ellagic acid by the method for chemosynthesis.Ellagic acid is the dimerization derivative of gallic acid, is main raw material with the gallic acid, selects suitable oxygenant for use, controls certain condition and carries out oxidative coupling and can obtain ellagic acid, and in this process, the productive rate of ellagic acid is about 20%-30%.In the methanol solution of methyl gallate, add saturated barium hydroxide solution, use the diluted acid acidifying behind the blowing air 2.5h, can obtain ellagic acid.Productive rate is about 30%; Methyl gallate is dissolved in 10% the ammoniacal liquor, continues to import air it is carried out oxidation, also can obtain ellagic acid.But, can cause detrimentally affect to environment owing to used organic reagent.
So, invent a kind of simple possible, do not cause the extracting method of the ellagic acid of environmental pollution to have good realistic meaning.
Summary of the invention
The object of the invention is to provide a kind of method of extracting ellagic acid in the pomegranate peel residue with biological enzyme.The principle of the inventive method is to extract solvent with water for being raw material with the pomegranate peel residue, behind the employing compound bio-enzyme enzymolysis Ellagitannins, uses the alkali lye enzymolysis reaction, forms the buck system simultaneously and extracts ellagic acid.Realization reduces cost, utilization of waste material, environmental friendliness, the purpose of raising ellagic acid output.
A kind of method with ellagic acid in the biological enzyme extraction pomegranate peel residue, carry out according to following operation steps:
(1) extraction of Ellagitannins
Oven dry and the pomegranate peel residue pulverized are placed water, and wherein, the mass ratio of pomegranate peel residue and water is 1: 15-1: 30, and be supersound process 2-3 time under the ultrasonic wave effect of 300-350W then with it at power, 20-45 minute at every turn, obtain the Ellagitannins aqueous solution;
(2) enzyme digestion reaction
The pH to 5.0-5.5 of the Ellagitannins aqueous solution that regulating step (1) obtains joined compound bio-enzyme in the above-mentioned Ellagitannins aqueous solution then, at 40-50 ℃ of following enzyme digestion reaction 1-2 hour;
(3) extraction of ellagic acid
Behind the enzyme digestion reaction, the enzymolysis solution pH to 10-14 of regulating step (2) carries out centrifugally then, and the supernatant liquor that obtains is the thick product of ellagic acid.
The enzyme amount that described compound bio-enzyme consumption needs in every gram Ellagitannins in the above-mentioned Ellagitannins aqueous solution of degrading, its component and consumption are: cellulase A:1.56-15.6U/g, cellulase B:0.05-0.5U/g, lytic enzyme: 0.2-2U/g.
Described cellulase A is a zytase.
Described cellulase B is that pentosanase is or/and the plant essence is carried prozyme SPE-007.
Described lytic enzyme is tannase and/or composite plant lytic enzyme.
All use NaOH regulator solution pH value in described step (2) and (3).
Use the Ellagitannins content of the Ellagitannins aqueous solution that the method for step (1) obtains to be 5.62-13.28g/L.
In the thick product of ellagic acid that described step (3) obtains, in the Ellagitannins content that obtains in the step (1), ellagic acid content is 12.69-24%.
Advantage of the present invention comprises following three aspects: the catalytic efficiency height of (1) compound bio-enzyme, and selectivity is strong, has shortened the production time; (2) use the alkali lye enzymolysis reaction, form the buck system simultaneously and extract ellagic acid, process is continuous, and technology is simple; (3) leaching process is a solvent with water fully, has realized the environmental friendliness of production process, and three waste discharge is few, is fit to large-scale commercial production.
China is pomegranate plantation big country, and its pericarp is used for the Chinese medicine medicinal material except that part, and major part is abandoned by people, causes very big waste.The present invention is a raw material with the pomegranate peel residue, and raw materials cost is low, can improve additional value of farm products.
Embodiment
Further specifying the present invention below by embodiment is raw material with the pomegranate peel residue, prepares the method for ellagic acid with compound bio-enzyme, but does not limit the present invention in the described scope of embodiments.
The amount that every gram Ellagitannins needs in the Ellagitannins aqueous solution that each enzyme dosage obtains in degradation step (1) in the compound bio-enzyme that uses in following examples.The raw material pomegranate peel residue adopts ultrasonic apparatus to carry out ultrasonication.
Embodiment 1:
(1) extraction of Ellagitannins
After the pomegranate rind of oven dry pulverized, be added to the water, wherein the mass ratio of pomegranate peel residue and water is 1: 15, and placing power is supersound process 3 times under the ultrasonic wave effect of 300W, each 45 minutes, obtains the aqueous solution of Ellagitannins.Adopt forint-phenol colorimetry (Foline-Phenol) to measure Ellagitannins content in the above-mentioned Ellagitannins aqueous solution.The content of Ellagitannins is 10.19g/L in the Ellagitannins aqueous solution that obtains.
(2) enzyme digestion reaction
The Ellagitannins aqueous solution that step (1) is obtained, regulate pH to 5.0 with the NaOH aqueous solution, then successively to wherein adding zytase 158.96U (enzyme dosage of every gram Ellagitannins of promptly degrading is 15.6U), pentosanase 5.1U (enzyme dosage of every gram Ellagitannins of promptly degrading is 0.5U) and tannase 20.4U (enzyme dosage of every gram Ellagitannins of promptly degrading is 2U), at 50 ℃ of following water-bath concussions, enzyme digestion reaction 2 hours.
(3) extraction of ellagic acid
The enzymolysis solution that step (2) obtains is regulated pH to 10 with the NaOH aqueous solution, carry out centrifugal, centrifugal 20 minutes of 5000r/min, the supernatant liquor that obtains is the thick product of ellagic acid, through high-performance liquid chromatogram determination ellagic acid content, the extraction yield that obtains ellagic acid is 7.89% (in Ellagitannins content in the Ellagitannins aqueous solution that obtains in the step (1)).
Embodiment 2:
(1) extraction of Ellagitannins
After the pomegranate rind of oven dry pulverized, be added to the water, wherein the volume ratio of the quality of pomegranate peel residue and water is 1: 20, and placing power is supersound process 3 times under the ultrasonic wave effect of 330W, each 35 minutes, obtains the aqueous solution of Ellagitannins.Adopt forint-phenol colorimetry (Foline-Phenol) to measure Ellagitannins content in the above-mentioned Ellagitannins aqueous solution.The content of Ellagitannins is 8.90g/L in the Ellagitannins aqueous solution that obtains.
(2) enzyme digestion reaction
The Ellagitannins aqueous solution that step (1) is obtained, regulate pH to 5.3 with the NaOH aqueous solution, then successively to wherein adding zytase 92.56U (enzyme dosage of every gram Ellagitannins of promptly degrading is 10.4U), SPE-007 0.89U (enzyme dosage of every gram Ellagitannins of promptly degrading is 0.1U) and composite plant lytic enzyme 2.848U (enzyme dosage of every gram Ellagitannins of promptly degrading is 0.32U), at 45 ℃ of following water-bath concussions, enzyme digestion reaction 1 hour.
(3) extraction of ellagic acid
The enzymolysis solution that step (2) obtains is regulated pH to 13 with the NaOH aqueous solution, carry out centrifugal, centrifugal 20 minutes of 5000r/min, the supernatant liquor that obtains is the thick product of ellagic acid, through high-performance liquid chromatogram determination ellagic acid content, the extraction yield that obtains ellagic acid is 15.76% (in Ellagitannins content in the Ellagitannins aqueous solution that obtains in the step (1)).
Embodiment 3:
(1) extraction of Ellagitannins
After the pomegranate rind of oven dry pulverized, be added to the water, wherein the volume ratio of the quality of pomegranate peel residue and water is 1: 30, and placing power is supersound process 2 times under the ultrasonic wave effect of 350W, each 30 minutes, obtains the aqueous solution of Ellagitannins.Adopt forint-phenol colorimetry (Foline-Phenol) to measure Ellagitannins content in the above-mentioned Ellagitannins aqueous solution.The content of Ellagitannins is 5.62g/L in the Ellagitannins aqueous solution that obtains.
(2) enzyme digestion reaction
The Ellagitannins aqueous solution that step (1) is obtained, regulate pH to 5.5 with the NaOH aqueous solution, then successively to wherein adding zytase 8.767U (enzyme dosage of every gram Ellagitannins of promptly degrading is 1.56U), SPE-0070.281U (enzyme dosage of every gram Ellagitannins of promptly degrading is 0.05U) and tannase 1.124U (enzyme dosage of every gram Ellagitannins of promptly degrading is 0.2U), at 40 ℃ of following water-bath concussions, enzyme digestion reaction 1 hour.
(3) extraction of ellagic acid
The enzymolysis solution that step (2) obtains is regulated pH to 14 with the NaOH aqueous solution, carry out centrifugal, centrifugal 20 minutes of 5000r/min, the supernatant liquor that obtains is the thick product of ellagic acid, through high-performance liquid chromatogram determination ellagic acid content, the extraction yield that obtains ellagic acid is 24% (in Ellagitannins content in the Ellagitannins aqueous solution that obtains in the step (1)).
The used enzyme of above embodiment, zytase are available from Beijing Challenge Biotechnologies Co., Ltd., and enzyme is lived and is 13000U/g, SPE-007 and composite plant lytic enzyme are available from Novozymes Company, enzyme is lived and to be 100U/g, and tannase is available from permanent magnificent road, east, Nanning biotechnology limited liability company, and enzyme is lived and is 200U/g.
Claims (5)
1. the method with ellagic acid in the biological enzyme extraction pomegranate peel residue is characterized in that, carries out according to following operation steps:
(1) extraction of Ellagitannins
Oven dry and the pomegranate peel residue pulverized are placed water, and wherein, the mass ratio of pomegranate peel residue and water is 1: 15-1: 30, and be supersound process 2-3 time under the ultrasonic wave effect of 300-350W then with it at power, 20-45 minute at every turn, obtain the Ellagitannins aqueous solution;
(2) enzyme digestion reaction
The pH to 5.0-5.5 of the Ellagitannins aqueous solution that regulating step (1) obtains joined compound bio-enzyme in the above-mentioned Ellagitannins aqueous solution then, at 40-50 ℃ of following enzyme digestion reaction 1-2 hour;
(3) extraction of ellagic acid
Behind the enzyme digestion reaction, the enzymolysis solution pH to 10-14 of regulating step (2) carries out centrifugally then, and the supernatant liquor that obtains is the thick product of ellagic acid.
2. the method for extracting ellagic acid in the pomegranate peel residue with biological enzyme according to claim 1, it is characterized in that, the enzyme amount that the consumption of described compound bio-enzyme needs in every gram Ellagitannins in the above-mentioned Ellagitannins aqueous solution of degrading, its component and consumption are: cellulase A:1.56-15.6U/g, cellulase B:0.05-0.5U/g, lytic enzyme: 0.2-2U/g.
3. the method with ellagic acid in the biological enzyme extraction pomegranate peel residue according to claim 2 is characterized in that described cellulase A is a zytase.
4. the method with ellagic acid in the biological enzyme extraction pomegranate peel residue according to claim 2 is characterized in that described cellulase B is that pentosanase and/or plant essence are carried prozyme SPE-007.
5. the method with ellagic acid in the biological enzyme extraction pomegranate peel residue according to claim 2 is characterized in that described lytic enzyme is tannase and/or composite plant lytic enzyme.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453039A (en) * | 2010-11-03 | 2012-05-16 | 张守力 | method for extracting ellagic acid from pomegranate bark by enzyme hydrolysis method |
| CN104489854A (en) * | 2014-12-22 | 2015-04-08 | 贵州省健康茶科技有限公司 | Lagerstroemia speciosa preservative |
| CN106344633A (en) * | 2016-09-28 | 2017-01-25 | 江苏春申堂药业有限公司 | Preparation method of pericarpium granati extract under normal temperature |
| CN108434043A (en) * | 2018-06-12 | 2018-08-24 | 艾苛密（上海）健康科技股份有限公司 | Skin-whitening Punica granatum L. extract and preparation method thereof |
| CN113603704A (en) * | 2021-08-30 | 2021-11-05 | 湖南华诚生物资源股份有限公司 | Enzymatic preparation method for separating ellagic acid from byproducts of rubusoside production |
-
2009
- 2009-10-29 CN CN2009102369452A patent/CN101701234B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453039A (en) * | 2010-11-03 | 2012-05-16 | 张守力 | method for extracting ellagic acid from pomegranate bark by enzyme hydrolysis method |
| CN104489854A (en) * | 2014-12-22 | 2015-04-08 | 贵州省健康茶科技有限公司 | Lagerstroemia speciosa preservative |
| CN106344633A (en) * | 2016-09-28 | 2017-01-25 | 江苏春申堂药业有限公司 | Preparation method of pericarpium granati extract under normal temperature |
| CN108434043A (en) * | 2018-06-12 | 2018-08-24 | 艾苛密（上海）健康科技股份有限公司 | Skin-whitening Punica granatum L. extract and preparation method thereof |
| CN108434043B (en) * | 2018-06-12 | 2021-04-09 | 艾苛密（上海）健康科技股份有限公司 | Pomegranate extract for skin whitening and preparation method thereof |
| CN113603704A (en) * | 2021-08-30 | 2021-11-05 | 湖南华诚生物资源股份有限公司 | Enzymatic preparation method for separating ellagic acid from byproducts of rubusoside production |
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