CN102229973A - Extraction method of Chinese pine pollen small peptides - Google Patents
Extraction method of Chinese pine pollen small peptides Download PDFInfo
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- CN102229973A CN102229973A CN2011101613428A CN201110161342A CN102229973A CN 102229973 A CN102229973 A CN 102229973A CN 2011101613428 A CN2011101613428 A CN 2011101613428A CN 201110161342 A CN201110161342 A CN 201110161342A CN 102229973 A CN102229973 A CN 102229973A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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Abstract
The invention relates to an extraction method of Chinese pine pollen small peptides. The extraction method comprises the following steps of (1) mixing Chinese pine pollen and water, carrying out a wall breaking process on the Chinese pine pollen in a wet way through a colloid mill, and filtering, (2) extracting a soluble component from the Chinese pine pollen treated by the wall breaking process through adopting a solvent oil, (3) putting the Chinese pine pollen treated by the step 3 into a container with a stirrer, adding filtrate obtained from the step 1 into the Chinese pine pollen, adjusting a pH of the mixture to a pH of 8 to 9, and stirring the mixture for 2 to 4 hours at a temperature of 20 to 70 DEG C to obtain Chinese pine pollen mixed solution, and (4) carrying out processes of enzymatic hydrolysis and enzyme destruction, carrying out a centrifugal separation process, and removing residues to obtain nutrient solution rich in small peptides. The extraction method has a short production period and a high product yield. Nutrient components of extract produced by the extraction method are micro-molecule peptides which can be absorbed by human bodies easily. Active micro-molecule peptide nutrient solution prepared by the extraction method has a high water-solubility and can be used directly.
Description
Technical field
The invention belongs to molecular bioengineering and biochemical engineering field, the extracting method of the little peptide of especially a kind of Chinese pine pollen.
Background technology
Chinese pine pollen contains whole nutritive ingredients of needed by human body, and collocation rationally, and the nutrition of can all replenish, balancing human body is required have the title of " green gold ".Studies show that in recent years, hydrolysis end product major part is the little peptide that 2 or 3 amino-acid residues are formed under the digestive ferment effect of protein in animal digestive tract, it is fast that the absorption of peptide has speed, it is low to consume energy, be difficult for transporting between the saturated and various peptides characteristics of uncontested property and inhibition, the bio-enzyme engineering working method is under mild reaction conditions, hydrolysis generates materials such as polypeptide and amino acid, have very high nutritive value and functional performance, it is controlled to have reaction, side reaction is few, good product quality and do not destroy characteristics such as amino acid, biological enzyme will be that the plant protein resource development and use are the most effective, the most promising working method.
The Chinese pine pollen broken wall generally adopts air-flow dry method method, the energy consumption height, and the nutrition loss is big, have bibliographical information to adopt Wet-process wall breaking more satisfactory, but need to add grinding aid, and mashed prod needs drying behind the broken wall.The water yield when advantage of method of the present invention is Wet-process wall breaking improves several times, has avoided material thickness problem in the process of lapping, and the filtrate after the solid-liquid separation can be used as media when enzymic hydrolysis prepares little peptide.
Patent documentation CN101100691 discloses a kind of method of extracting little peptide with prozyme from Chinese pine pollen, because the auditor thinks that this application does not prove absolutely the dissolving-out method of Chinese pine pollen, therefore be considered to disclose insufficient, on the basis of the application based on this application, prove absolutely the wall-breaking method of Chinese pine pollen, and obtained good effect.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art part, the extracting method of the little peptide of a kind of Chinese pine pollen is provided, and present method is to adopt the wet colloid mill broken wall, and the water of adding can be used for the medium of zymyhydrolyzed protein matter, this method is not only with short production cycle, the product yield height.
The extracting method of the little peptide of a kind of Chinese pine pollen is characterized in that: the method for extracting little peptide may further comprise the steps:
(1) Chinese pine pollen is mixed with water, with colloidal mill Chinese pine pollen is carried out Wet-process wall breaking, the broken wall after-filtration obtains broken wall Chinese pine pollen and filtered liquid;
(2) the broken wall Chinese pine pollen is gone out the composition of solubility with solvent extraction;
(3) put into the container that has agitator behind the Chinese pine pollen desolventizing after will extracting, add filtered liquid, after regulator solution pH value is 8～9, under 20～70 ℃ of conditions, stirred 2～4 hours, formation Chinese pine pollen mixed solution;
(4) in the Chinese pine pollen mixed solution, add prozyme, fully stirred 2～8 hours;
The enzyme that goes out under (5) 90 ℃ of conditions, centrifugation are removed the material slag, promptly obtain small peptide nutrient liquid.
And the solvent of described step (2) is edible solvent oil or supercritical CO
2
And the mass ratio of described Chinese pine pollen and water is 1: 5～15.
And the described enzyme that adds in the Chinese pine pollen mixed solution is the wherein several mixture in Sumizyme MP, neutral protease, papoid, the trypsinase.
And the weight ratio of the addition of described enzyme and Chinese pine pollen butt is 0.2～0.4: 50～150.
And the medium and small peptide molecular weight of described small peptide nutrient liquid is 300-1000 dalton, i.e. 2～8 small-molecular peptides that amino acid is formed.
Advantage of the present invention and beneficial effect are:
1. the present invention utilizes wet method with the Chinese pine pollen broken wall, the filtrate of containing nutrition composition behind the broken wall can be used for the enzyme hydrolysis process media, when having overcome Wet-process wall breaking because the little material thickness of solid-liquid ratio, the high problem of the big subsequent disposal expense of solid-liquid ratio keeps to greatest extent that Chinese pine pollen is natural, the character of high nutrition.
2. the Chinese pine pollen extracting solution of taking method of the present invention to make because broken wall is thorough, makes the abundant strippings of nutritive ingredient such as active small-molecular peptides and VITAMIN, polysaccharide, mineral substance, edibility more, and nutrition is abundanter.
3. the Chinese pine pollen extracting solution separate easily that adopts the present invention to make, water-soluble height, do not destroy the nutrition of pollen, and do not exist pollen extracting liquid to be subjected to the protein coagulated phenomenon of thermogenesis, this product has micronutrient elements such as the active small peptide of the absorption of human body of being easy to and some mineral substance.
4. the present invention utilizes combinative enzyme hydrolysis Chinese pine pollen albumen, reaction characteristics according to different enzymes, need not regulate the pH value, because protein produces more carboxyl in hydrolytic process, system pH is constantly reduced, initial system is an alkalescence, the Sumizyme MP that adopts 2709 fermenting bacillus subtilis to obtain, be suitable for most the pH value for 9-11, main catalytic serine is when system pH drops to 8 when following, neutral protease plays a role, neutral protease is obtained by fermentation of bacillus subtilis, is suitable for the pH value most and is 6.0-7.5, can be owing to the multiple proteins hydrolysis; And papoid has the pH scope of broad, owing to regulate the pH value without alkali, system does not produce salt.
Embodiment
The invention will be further described below by specific embodiment, and following examples are descriptive, is not determinate, can not limit protection scope of the present invention with this.
Embodiment 1:
The present invention extracts the method for little peptide with enzyme from Chinese pine pollen, be the catalyzed reaction effect that utilizes prozyme, makes the protein transduction in the Chinese pine pollen become the active small molecular peptide that easily is absorbed by the body, and its extracting method may further comprise the steps:
(1). add after the removal of impurities 200 parts of Chinese pine pollens in open container, 2000 parts of tap water grind broken wall with colloidal mill, examine under a microscope broken wall and reach at 95% o'clock, separate with plate-and-frame filter press, obtain broken wall Chinese pine pollen and filtrate;
(2). get Chinese pine pollen behind 150 parts of broken walls, extract the non-protein ingredient of Chinese pine pollen solubility with carbon dioxide upercritical fluid extraction; Chinese pine pollen part after the extraction is put into the container of a belt stirrer, and under 45 ℃ of temperature condition, the filtrate that adds 1800 parts is made mixed solution, regulates pH of mixed=10 with alkali, soaking and stirring 3 hours.
(3). in above-mentioned mixed solution, add 0.2 part of Sumizyme MP (permanent magnificent road, east, Nanning biotechnology limited liability company, bacillus alkaline protease), 0.2 part neutral protease (east, Nanning permanent magnificent road biotechnology limited liability company), under 45 ℃ of temperature condition, fully stirred 3 hours, reaction finishes, pH=5.8;
(4). the enzyme 30min that under 90 ℃ of conditions, goes out, be cooled to 40 ℃ of centrifugations, remove the material slag, make small peptide nutrient liquid.
(5). adopt the high effective liquid chromatography for measuring molecular weight, mainly go out the peak position, be i.e. 2-8 the little peptide that amino acid is formed 300-700 dalton.
Embodiment 2:
PH value in the processing step among the embodiment 1 (2) is adjusted into 9, makes the kind of enzyme in the processing step (3) into papoid by neutral protease and get final product reaction end, system pH=5.8.
Other working method are with embodiment 1.
Embodiment 3
(1). add after the removal of impurities 200 parts of Chinese pine pollens in open container, 1000 parts of tap water grind broken wall with colloidal mill, examine under a microscope broken wall and reach at 95% o'clock, separate with plate-and-frame filter press, obtain broken wall Chinese pine pollen and filtrate;
(2). get Chinese pine pollen behind 100 parts of broken walls, extract the non-protein ingredient of Chinese pine pollen solubility with 6# solvent oil (meeting GB 16629-2008 standard); Chinese pine pollen after the extraction is put into the container of a belt stirrer, under 50 ℃ of temperature condition, solution is adjusted to pH=10, soaking and stirring 3 hours with 1000 parts filtrates.
(3). in above-mentioned solution, add 0.2 part of Sumizyme MP, 0.1 part of neutral protease, 0.2 part of papoid fully stirred 3 hours, and reaction finishes, pH=5.5;
(4). the enzyme 30min that under 90 ℃ of conditions, goes out, be cooled to 40 ℃ of centrifugations, the material slag is removed in centrifugation, makes small peptide nutrient liquid.
Claims (6)
1. the extracting method of the little peptide of Chinese pine pollen, it is characterized in that: the method for extracting little peptide may further comprise the steps:
(1) Chinese pine pollen is mixed with water, with colloidal mill Chinese pine pollen is carried out Wet-process wall breaking, the broken wall after-filtration obtains broken wall Chinese pine pollen and filtered liquid;
(2) the broken wall Chinese pine pollen is gone out the composition of solubility with solvent extraction;
(3) put into the container that has agitator behind the Chinese pine pollen desolventizing after will extracting, add filtered liquid, after regulator solution pH value is 8～9, under 20～70 ℃ of conditions, stirred 2～4 hours, formation Chinese pine pollen mixed solution;
(4) in the Chinese pine pollen mixed solution, add prozyme, fully stirred 2～8 hours;
The enzyme that goes out under (5) 90 ℃ of conditions, centrifugation are removed the material slag, promptly obtain small peptide nutrient liquid.
2. the extracting method of the little peptide of Chinese pine pollen according to claim 1 is characterized in that: the solvent of described step (2) is edible solvent oil or supercritical CO
2
3. the extracting method of the little peptide of Chinese pine pollen according to claim 1 is characterized in that: the mass ratio of described Chinese pine pollen and water is 1: 5～15.
4. the extracting method of the little peptide of Chinese pine pollen according to claim 1 is characterized in that: the described enzyme that adds in the Chinese pine pollen mixed solution is the wherein several mixture in Sumizyme MP, neutral protease, papoid, the trypsinase.
5. the extracting method of the little peptide of Chinese pine pollen according to claim 1 is characterized in that: the weight ratio of the addition of described enzyme and Chinese pine pollen butt is 0.2～0.4: 50～150.
6. the extracting method of the little peptide of Chinese pine pollen according to claim 1 is characterized in that: the medium and small peptide molecular weight of described small peptide nutrient liquid is 300-1000 dalton, i.e. 2～8 small-molecular peptides that amino acid is formed.
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| CN2011101613428A CN102229973B (en) | 2011-06-15 | 2011-06-15 | Extraction method of Chinese pine pollen small peptides |
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Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659917A (en) * | 2012-05-24 | 2012-09-12 | 烟台新时代健康产业有限公司 | Method for separating and preparing crude protein and enzymolysis peptide from pollen pini |
| CN103245557A (en) * | 2013-05-09 | 2013-08-14 | 甘肃蓝科石化高新装备股份有限公司 | Double-tube charging/discharging material distribution device for single-unit and double-tube hydrostatic testing machine |
| CN103392904A (en) * | 2013-08-02 | 2013-11-20 | 江苏丘陵地区镇江农业科学研究所 | Method used for extracting black soya bean polypeptide by combining wet ball milling with enzyme method |
| CN104593461A (en) * | 2014-12-29 | 2015-05-06 | 唯美度科技（北京）有限公司 | Low-molecular-weight bee-pollen protein peptide and preparation method thereof |
| CN106261972A (en) * | 2016-08-22 | 2017-01-04 | 得利斯集团有限公司 | A kind of extracting method of Carnis Mactrae peptide |
| CN106359839A (en) * | 2016-08-22 | 2017-02-01 | 得利斯集团有限公司 | Extraction method of oyster peptides |
| CN107252099A (en) * | 2017-06-27 | 2017-10-17 | 烟台新时代健康产业有限公司 | A kind of health food for improving osteoarthritic inflammation and preparation method thereof |
| CN107523599A (en) * | 2017-09-01 | 2017-12-29 | 兰溪市沉默生物科技有限公司 | The method that fermentation method prepares peony pollen polypeptide |
| CN107988299A (en) * | 2017-12-01 | 2018-05-04 | 烟台新时代健康产业有限公司 | A kind of method that anti-oxidant pollen pini peptide is prepared using the pollen pini dregs of rice |
| CN109320358A (en) * | 2018-12-03 | 2019-02-12 | 孙宝泉 | A kind of Selenium-rich nutrient solution and its application in tree peony grafting culture |
| CN110229865A (en) * | 2019-07-11 | 2019-09-13 | 云南惠本生物工程有限公司 | A kind of pollen pini active peptide extracting method |
| CN111621538A (en) * | 2020-06-22 | 2020-09-04 | 浙江亚林生物科技股份有限公司 | Preparation method and application of pollen pini oligopeptide |
| CN114081854A (en) * | 2021-11-19 | 2022-02-25 | 烟台新时代健康产业日化有限公司 | Pollen pini extract with barrier repair function and fermentation preparation process thereof |
| CN116333820A (en) * | 2023-03-15 | 2023-06-27 | 烟台新时代健康产业有限公司 | Method for extracting pollen Pini essential oil from pollen Pini without solvent |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659917A (en) * | 2012-05-24 | 2012-09-12 | 烟台新时代健康产业有限公司 | Method for separating and preparing crude protein and enzymolysis peptide from pollen pini |
| CN103245557A (en) * | 2013-05-09 | 2013-08-14 | 甘肃蓝科石化高新装备股份有限公司 | Double-tube charging/discharging material distribution device for single-unit and double-tube hydrostatic testing machine |
| CN103392904A (en) * | 2013-08-02 | 2013-11-20 | 江苏丘陵地区镇江农业科学研究所 | Method used for extracting black soya bean polypeptide by combining wet ball milling with enzyme method |
| CN104593461A (en) * | 2014-12-29 | 2015-05-06 | 唯美度科技（北京）有限公司 | Low-molecular-weight bee-pollen protein peptide and preparation method thereof |
| CN106261972A (en) * | 2016-08-22 | 2017-01-04 | 得利斯集团有限公司 | A kind of extracting method of Carnis Mactrae peptide |
| CN106359839A (en) * | 2016-08-22 | 2017-02-01 | 得利斯集团有限公司 | Extraction method of oyster peptides |
| CN107252099A (en) * | 2017-06-27 | 2017-10-17 | 烟台新时代健康产业有限公司 | A kind of health food for improving osteoarthritic inflammation and preparation method thereof |
| CN107523599A (en) * | 2017-09-01 | 2017-12-29 | 兰溪市沉默生物科技有限公司 | The method that fermentation method prepares peony pollen polypeptide |
| CN107988299A (en) * | 2017-12-01 | 2018-05-04 | 烟台新时代健康产业有限公司 | A kind of method that anti-oxidant pollen pini peptide is prepared using the pollen pini dregs of rice |
| CN107988299B (en) * | 2017-12-01 | 2021-06-01 | 烟台新时代健康产业有限公司 | Method for preparing antioxidant pollen pini peptide by utilizing pollen pini dregs |
| CN109320358A (en) * | 2018-12-03 | 2019-02-12 | 孙宝泉 | A kind of Selenium-rich nutrient solution and its application in tree peony grafting culture |
| CN110229865A (en) * | 2019-07-11 | 2019-09-13 | 云南惠本生物工程有限公司 | A kind of pollen pini active peptide extracting method |
| CN111621538A (en) * | 2020-06-22 | 2020-09-04 | 浙江亚林生物科技股份有限公司 | Preparation method and application of pollen pini oligopeptide |
| CN114081854A (en) * | 2021-11-19 | 2022-02-25 | 烟台新时代健康产业日化有限公司 | Pollen pini extract with barrier repair function and fermentation preparation process thereof |
| CN114081854B (en) * | 2021-11-19 | 2023-08-15 | 烟台新时代健康产业日化有限公司 | Pollen pini extract with barrier repairing function and fermentation preparation process thereof |
| CN116333820A (en) * | 2023-03-15 | 2023-06-27 | 烟台新时代健康产业有限公司 | Method for extracting pollen Pini essential oil from pollen Pini without solvent |
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