CN111978430A - Preparation method of oat beta-glucan - Google Patents
Preparation method of oat beta-glucan Download PDFInfo
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- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
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Abstract
The invention discloses a preparation method of oat beta-glucan, which comprises the steps of firstly carrying out degreasing treatment on raw materials, facilitating the subsequent dissolution of effective components and the reduction of impurities, and then carrying out electrolysis treatment on mixed liquid after water extraction to promote the separation and dissolution of the beta-glucan in oat bran; after the electrolysis treatment, the ultrasonic treatment and the enzymolysis treatment are carried out to promote the complete release of the beta-glucan, the extraction rate of the beta-glucan is effectively improved, impurities are removed through isoelectric point precipitation, flocculation precipitation and alcohol precipitation in the extraction process to ensure the purity of the product, finally, separation membranes of different specifications are adopted to carry out series separation to obtain products of different molecular weights, the requirements of various fields are met, finally, the chemical structure of the beta-glucan can be furthest prevented from being damaged through a vacuum drying mode, the excellent chemical activity of the beta-glucan is kept, and the practical application value is high.
Description
Technical Field
The invention relates to the technical field of fine chemical engineering, and particularly relates to a preparation method of oat beta-glucan.
Background
Oats have a comprehensive and higher nutritional value than other cereals. Five nutritional indexes such as protein, fat, vitamins, mineral elements, cellulose and the like are in the first place. Research shows that the oat contains a large amount of dietary fiber, wherein the water-soluble dietary fiber beta-glucan has higher application value. The beta-glucan is non-starch viscous polysaccharide formed by connecting a pyran glucose unit through 1-3 and 1-4 glycosidic bonds, has special physiological functions of reducing cholesterol, regulating blood sugar, improving immunity and the like, prevents and treats constipation, improves intestinal microenvironment, is a cosmetic intermediate and the like, and is widely applied to the fields of medicines, health-care foods and cosmetics.
In the prior art, oat bran is mostly adopted to extract beta-glucan, starch and protein are hydrolyzed by an enzymolysis method in the extraction process, then the beta-glucan is separated from filtrate, and in the process of extracting the beta-glucan by utilizing the oat bran, because the oat bran has certain grease and other substances, the dissolution and the purity of the beta-glucan are further influenced, a certain organic solvent or ethanol is mostly added in the extraction process to dissolve out the other substances and then purification is carried out, however, in the prior art, the removal and purification of the substances such as starch, cellulose and the like are mostly aimed at, partial organic solvent or ethanol and other substances can be remained in the purified beta-glucan, the extraction cost of the beta-glucan is high, the purity is limited, the extraction rate is not good, and the application in different fields has certain requirements on the molecular weight of the beta-glucan, however, the lack of a scheme for preparing beta-glucan with a specific molecular weight integrally in the prior art severely limits the development and application of the beta-glucan.
Therefore, the problem to be solved by those skilled in the art is how to provide a method for preparing oat β -glucan with high efficiency, high purity and controllable molecular weight.
Disclosure of Invention
In view of the above, the invention provides a preparation method of oat beta-glucan, which improves and integrates an extraction process and efficiently and quickly produces high-purity oat beta-glucan.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of oat beta-glucan comprises the following steps:
(1) dehydrating and drying oat grains, crushing to 10-150 meshes, separating to obtain oat bran, and then performing microwave drying on the oat bran for 5-10min in 20-60 kW;
(2) performing supercritical carbon dioxide extraction on the oat bran in the step (1) for 3-5 h;
(3) dispersing the oat bran treated in the step (2) into clear water, and then inserting a carbon rod electrode for electrolysis for 10-15 min;
(4) carrying out ultrasonic treatment on the electrolyzed mixed solution for 60-150min, then adjusting the temperature and the pH value, and adding mixed enzyme for enzymolysis to obtain an enzymolysis solution;
(5) adjusting pH of the enzymolysis solution to 8-10 at 20-95 deg.C, extracting for 30-90min and repeating the operation for 1-3 times;
(6) adjusting the pH of the extracting solution obtained in the step (5) to 3-6, heating to 80-100 ℃, heating for 30-90min, standing for precipitation for 1-12h, and separating to obtain a supernatant;
(7) on the basis of the quality of the supernatant, adding 0.01-2.0 wt% of chitosan into the supernatant obtained in the step (6), then adding 0.1-5.0 wt% of sodium alginate, uniformly stirring, standing for precipitation for 1-15h, and separating to obtain a supernatant;
(8) and (4) filtering and separating the supernatant obtained in the step (7) by using ultrafiltration membranes with different molecular weights in a series membrane separation mode, collecting trapped liquids in sections, concentrating the trapped liquids respectively, cooling to room temperature, adding alcohol for alcohol precipitation, collecting precipitates, and drying the precipitates in vacuum to obtain the high-purity oat beta-glucan with different molecular weights.
Preferably, the supercritical carbon dioxide extraction pressure in the step (2) is 20-50MPa, the temperature is 40-60 ℃, and the extraction flow rate is 2200-.
The beneficial effects of the preferred technical scheme are as follows: the supercritical extraction can remove the grease in the oat bran firstly, does not damage the effective components in the oat bran, is beneficial to the dissolution of the subsequent beta-glucan, reduces the impurity content in the subsequent treatment process and is beneficial to the purification.
Preferably, the feed-liquid ratio of the oat bran to the clear water in the step (3) is 1 (10-25), the pH is adjusted to 3-4, and the electrolytic voltage is 15-20V.
Preferably, the ultrasonic treatment temperature in the step (4) is 40-60 ℃ and the time is 60-150 min.
The beneficial effects of the preferred technical scheme are as follows: the reaction can be promoted to move in the positive direction within the range of the feed-liquid ratio, so that the beta-glucan can be fully dissolved out, the mixed solution is subjected to electrolysis treatment, and the rapid directional movement of ions in the mixed solution can be effectively promoted, so that the effective impact can be carried out on the tissue components of the oat bran, the decomposition of the tissue of the oat bran is promoted, and the separation and the dissolution of the beta-glucan in the oat bran are promoted;
and the activity of endogenous beta-glucanase in oat bran tissues can be reduced by carrying out ultrasonic treatment after electrolytic treatment, and during ultrasonic treatment, the beta-glucan can absorb water, so that the swelling and cracking of cell walls are promoted, the release of the beta-glucan combined with proteins, pentosan, hemicellulose and other substances in an aleurone layer is promoted, the solubility of the beta-glucanase is improved, and the extraction rate of the beta-glucan is effectively improved.
Preferably, the temperature is adjusted to 30-40 ℃ and the pH is 6-8 before enzymolysis in the step (4), the added mixed enzymes are alpha-amylase, saccharifying enzyme and alkaline protease, the addition amount of the alpha-amylase is 0.1-0.3 wt%, the addition amount of the saccharifying enzyme is 0.5-3 wt% and the addition amount of the alkaline protease is 1-8 wt% based on the weight of the oat bran.
The beneficial effects of the preferred technical scheme are as follows: the temperature and the pH value of the system are adjusted to achieve the optimal enzymolysis condition, the beta-glucan combined in the starch and the protein is further and completely released through the enzymolysis of the complex enzyme, and the starch and the protein are hydrolyzed into small molecules to be beneficial to completely removing the small molecules through isoelectric point precipitation and flocculation precipitation in the subsequent process.
Preferably, the cut-off molecular weight of the ultrafiltration membrane in the step (8) is 30000-1500000Da respectively.
The beneficial effects of the preferred technical scheme are as follows: within the range, the molecular weight of the membrane can be selected according to requirements, so that the membrane is suitable for different product requirements.
Preferably, the concentration in the step (8) adopts double-effect concentration, the concentration temperature is 40-70 ℃, the vacuum degree is 40-80Kpa, the time is 1-10h, and the concentration of the concentrated solution is 20-40 DEG Be.
Preferably, the alcohol is cooled to 20-30 ℃ before alcohol precipitation in the step (8), the volume concentration of the added alcohol is 60-90%, and the adding amount is 4-6 times of the volume of the concentrated solution.
Preferably, in the step (8), the vacuum drying temperature is 50-80 ℃, the vacuum degree is 60-100KPa, and the drying time is 2-8 h.
The beneficial effects of the preferred technical scheme are as follows: by adopting a vacuum drying mode, the drying temperature is effectively controlled, the chemical structure of the beta-glucan can be furthest prevented from being damaged, and the excellent chemical activity of the beta-glucan is kept.
Through the technical scheme, compared with the prior art, the preparation method of the oat beta-glucan can greatly improve the extraction rate and purity of the beta-glucan, firstly, the raw material is degreased, the subsequent dissolution of effective components and the reduction of impurities are facilitated, and then the separation and dissolution of the beta-glucan in the oat bran are promoted by performing electrolytic treatment on the mixed solution after water extraction; after the electrolysis treatment, the ultrasonic treatment and the enzymolysis treatment are carried out to promote the complete release of the beta-glucan, the extraction rate of the beta-glucan is effectively improved, impurities are removed through isoelectric point precipitation, flocculation precipitation and alcohol precipitation in the extraction process to ensure the purity of the product, finally, separation membranes of different specifications are adopted to carry out series separation to obtain products of different molecular weights, the requirements of various fields are met, finally, the chemical structure of the beta-glucan can be furthest prevented from being damaged through a vacuum drying mode, the excellent chemical activity of the beta-glucan is kept, and the practical application value is high.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
(1) Dehydrating and drying oat grains, crushing the oat grains to 100 meshes, separating to obtain oat bran, and then performing microwave drying on the oat bran for 10min in 20kW mode;
(2) performing supercritical carbon dioxide extraction on the oat bran in the step (1) under the conditions of pressure of 20MPa and temperature of 60 ℃, wherein the extraction time is 5h, and the extraction flow is 4300L/h;
(3) dispersing the oat bran treated in the step (2) into clear water according to the feed-liquid ratio of 1:10, adjusting the pH to 3, inserting a carbon rod electrode, and electrolyzing for 15min at 15V;
(4) performing ultrasonic treatment on the electrolyzed mixed solution at 60 ℃ for 60min, then adjusting the temperature to be 30 ℃ and the pH value to be 6, and adding mixed enzyme for enzymolysis to obtain an enzymolysis solution, wherein the mixed enzyme comprises alpha-amylase, saccharifying enzyme and alkaline protease, and the weight of oat bran is taken as a basis, the adding amount of the alpha-amylase is 0.1 wt%, the saccharifying enzyme is 0.5 wt%, and the alkaline protease is 1 wt%;
(5) adjusting pH of the enzymolysis solution to 8, extracting at 30 deg.C for 90min, and repeating the operation for 3 times;
(6) adjusting the pH of the extracting solution obtained in the step (5) to 3, heating to 80 ℃, heating for 90min, standing for precipitation for 6h, and separating to obtain a supernatant;
(7) on the basis of the quality of the supernatant, adding 2.0 wt% of chitosan into the supernatant obtained in the step (6), then adding 5.0 wt% of sodium alginate, then uniformly stirring, standing for precipitation for 6 hours, and separating to obtain the supernatant;
(8) filtering and separating the supernatant obtained in the step (7) by ultrafiltration membranes with different molecular weight cut-off in a series membrane separation mode, collecting cut-off liquid in sections, and performing double-effect concentration on the cut-off liquid respectively, wherein the concentration temperature is 40 ℃, the vacuum degree is 80Kpa, the time is 3 hours, and the concentration of the concentrated liquid is 20 degrees Be; cooling to room temperature after concentration, adding alcohol with concentration of 60% -90% and 4 times of the volume of the concentrated solution for alcohol precipitation, collecting precipitate, and vacuum drying the precipitate at 50 ℃ under the vacuum degree of 100KPa to obtain high-purity oat beta-glucan with different molecular weights.
Wherein the cut-off molecular weight of the ultrafiltration membrane in the step (8) is 30000-1500000Da respectively.
Example 2
(1) Dehydrating and drying oat grains, crushing the oat grains to 100 meshes, separating to obtain oat bran, and then drying the oat bran for 8min by using 40kW microwave;
(2) performing supercritical carbon dioxide extraction on the oat bran in the step (1) under the conditions of the pressure of 35MPa and the temperature of 50 ℃, wherein the extraction time is 5h, and the extraction flow is 3500L/h;
(3) dispersing the oat bran treated in the step (2) into clear water according to the feed-liquid ratio of 1:20, adjusting the pH to 4, inserting a carbon rod electrode, and electrolyzing for 10min at 20V;
(4) performing ultrasonic treatment on the electrolyzed mixed solution at 50 ℃ for 100min, then adjusting the temperature to be 30 ℃ and the pH value to be 7, and adding mixed enzyme for enzymolysis to obtain an enzymolysis solution, wherein the mixed enzyme comprises alpha-amylase, saccharifying enzyme and alkaline protease, and the weight of oat bran is taken as a basis, the adding amount of the alpha-amylase is 0.2 wt%, the saccharifying enzyme is 1.8 wt%, and the alkaline protease is 5 wt%;
(5) adjusting pH of the enzymolysis solution to 9, extracting at 550 deg.C for 50min, and repeating the operation for 3 times;
(6) adjusting the pH of the extracting solution obtained in the step (5) to 5, heating to 90 ℃, heating for 60min, standing for precipitation for 12h, and separating to obtain a supernatant;
(7) on the basis of the quality of the supernatant, adding 0.01 wt% of chitosan into the supernatant obtained in the step (6), then adding 5.0 wt% of sodium alginate, then uniformly stirring, standing for precipitation for 15h, and separating to obtain the supernatant;
(8) filtering and separating the supernatant obtained in the step (7) by ultrafiltration membranes with different molecular weight cut-off in a series membrane separation mode, collecting cut-off liquid in sections, and performing double-effect concentration on the cut-off liquid respectively, wherein the concentration temperature is 550 ℃, the vacuum degree is 60Kpa, the time is 5 hours, and the concentration of the concentrated liquid is 30 DEG Be; cooling to room temperature after concentration, adding alcohol with the concentration of 60% -90% and the volume of 5 times of the concentrated solution, precipitating with ethanol, collecting the precipitate, and drying the precipitate in vacuum at 650 ℃ and the vacuum degree of 80KPa to obtain the high-purity oat beta-glucan with different molecular weights.
Wherein the cut-off molecular weight of the ultrafiltration membrane in the step (8) is 30000-1500000Da respectively.
Example 3
(1) Dehydrating and drying oat grains, crushing the oat grains to 150 meshes, separating to obtain oat bran, and then drying the oat bran for 5min by 60kW microwave;
(2) performing supercritical carbon dioxide extraction on the oat bran in the step (1) under the conditions of 50MPa of pressure and 40 ℃, wherein the extraction time is 3h, and the extraction flow rate is 2200L/h;
(3) dispersing the oat bran treated in the step (2) into clear water according to the feed-liquid ratio of 1:25, adjusting the pH to 4, inserting a carbon rod electrode, and electrolyzing for 15min at 20V;
(4) carrying out ultrasonic treatment on the electrolyzed mixed solution for 150min at 40 ℃, then adjusting the temperature to 40 ℃ and the pH value to 8, and adding mixed enzyme for enzymolysis to obtain an enzymolysis solution, wherein the mixed enzyme is alpha-amylase, saccharifying enzyme and alkaline protease, and the weight of oat bran is taken as a basis, the adding amount of the alpha-amylase is 0.3 wt%, the saccharifying enzyme is 3 wt%, and the alkaline protease is 8 wt%;
(5) adjusting pH of the enzymolysis solution to 10, extracting at 95 deg.C for 30min, and repeating the operation for 2 times;
(6) adjusting the pH of the extracting solution obtained in the step (5) to 6, heating to 100 ℃, heating for 30min, standing for precipitation for 12h, and separating to obtain a supernatant;
(7) on the basis of the quality of the supernatant, adding 2.0 wt% of chitosan into the supernatant obtained in the step (6), then adding 0.1 wt% of sodium alginate, then uniformly stirring, standing for precipitation for 12 hours, and separating to obtain the supernatant;
(8) filtering and separating the supernatant obtained in the step (7) by ultrafiltration membranes with different molecular weight cut-off in a series membrane separation mode, collecting cut-off liquid in sections, and performing double-effect concentration on the cut-off liquid respectively, wherein the concentration temperature is 70 ℃, the vacuum degree is 40Kpa, the time is 10 hours, and the concentration of the concentrated liquid is 40 degrees Be; cooling to room temperature after concentration, adding alcohol with concentration of 60% -90% 6 times of the volume of the concentrated solution for alcohol precipitation, collecting precipitate, and vacuum drying the precipitate at 50 ℃ under the vacuum degree of 60KPa to obtain high-purity oat beta-glucan with different molecular weights.
Wherein the cut-off molecular weight of the ultrafiltration membrane in the step (8) is 30000-
The yield of oat beta glucan was calculated as: the total amount of oat beta-glucan with each molecular weight obtained by extraction is 100% of the total weight of the raw material; the purity was determined by conventional means and the results were as follows:
| yield (%) | Purity (%) | |
| Example 1 | 7.96 | 91％ |
| Example 2 | 8.32 | 93％ |
| Example 3 | 8.21 | 89％ |
Therefore, by adopting the technical scheme of the invention, the high yield of the oat beta-glucan can be ensured, the purity of the product is greatly improved, impurities are effectively reduced, the limitation of the product to the application of the product in food or cosmetics due to the allergen doped in the product is avoided, and the high-purity oat beta-glucan with different molecular weights obtained by films with different specifications can be suitable for various fields and has higher practical application value.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (9)
1. The preparation method of oat beta-glucan is characterized by comprising the following steps:
(1) dehydrating and drying oat grains, crushing to 10-150 meshes, separating to obtain oat bran, and then performing microwave drying on the oat bran for 5-10min in 20-60 kW;
(2) performing supercritical carbon dioxide extraction on the oat bran in the step (1) for 3-5 h;
(3) dispersing the oat bran treated in the step (2) into clear water, and then inserting a carbon rod electrode for electrolysis for 10-15 min;
(4) carrying out ultrasonic treatment on the electrolyzed mixed solution for 60-150min, then adjusting the temperature and the pH value, and adding mixed enzyme for enzymolysis to obtain an enzymolysis solution;
(5) adjusting pH of the enzymolysis solution to 8-10 at 20-95 deg.C, extracting for 30-90min and repeating the operation for 1-3 times;
(6) adjusting the pH of the extracting solution obtained in the step (5) to 3-6, heating to 80-100 ℃, heating for 30-90min, standing for precipitation for 1-12h, and separating to obtain a supernatant;
(7) on the basis of the quality of the supernatant, adding 0.01-2.0 wt% of chitosan into the supernatant obtained in the step (6), then adding 0.1-5.0 wt% of sodium alginate, uniformly stirring, standing for precipitation for 1-15h, and separating to obtain a supernatant;
(8) and (4) filtering and separating the supernatant obtained in the step (7) by using ultrafiltration membranes with different molecular weights in a series membrane separation mode, collecting trapped liquids in sections, concentrating the trapped liquids respectively, cooling to room temperature, adding alcohol for alcohol precipitation, collecting precipitates, and drying the precipitates in vacuum to obtain the high-purity oat beta-glucan with different molecular weights.
2. The method for preparing oat beta-glucan according to claim 1, wherein the supercritical carbon dioxide extraction pressure in step (2) is 20-50MPa, the temperature is 40-60 ℃, and the extraction flow rate is 2200-.
3. The method for preparing oat beta-glucan according to claim 1, wherein the feed-liquid ratio of oat bran to clear water in the step (3) is 1 (10-25), the pH is adjusted to 3-4, and the electrolytic voltage is 15-20V.
4. The method for preparing oat beta-glucan according to claim 1, wherein the ultrasonic treatment temperature in the step (4) is 40-60 ℃ and the ultrasonic treatment time is 60-150 min.
5. The method for preparing oat beta-glucan according to claim 1, wherein the temperature is adjusted to 30-40 ℃ and the pH is 6-8 before enzymolysis in the step (4), the added mixed enzyme is alpha-amylase, saccharifying enzyme and alkaline protease, the alpha-amylase is added in an amount of 0.1-0.3 wt%, the saccharifying enzyme is 0.5-3 wt%, and the alkaline protease is 1-8 wt% based on the weight of oat bran.
6. The method for preparing oat beta-glucan according to claim 1, wherein the cut-off molecular weight of the ultrafiltration membrane in the step (8) is 30000-1500000Da respectively.
7. The method for preparing oat beta-glucan according to claim 1, wherein the concentration in the step (8) adopts double-effect concentration, the concentration temperature is 40-70 ℃, the vacuum degree is 40-80Kpa, the time is 1-10h, and the concentration of the concentrated solution is 20-40 ° Be.
8. The method for preparing oat beta-glucan according to claim 1, wherein the cooling to 20-30 ℃ is performed before the alcohol precipitation in the step (8), the volume concentration of the added alcohol is 60% -90%, and the added amount is 4-6 times of the volume of the concentrated solution.
9. The method for preparing oat beta-glucan according to claim 1, wherein the vacuum drying temperature in the step (8) is 50-80 ℃, the vacuum degree is 60-100KPa, and the drying time is 2-8 h.
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| CN115413712A (en) * | 2022-09-15 | 2022-12-02 | 草根知本集团有限公司 | Oat milk and preparation method thereof |
| CN115444139A (en) * | 2022-09-15 | 2022-12-09 | 草根知本集团有限公司 | Application of beta-glucan or extract thereof in preparation of food or medicine for improving hyperuricemia |
| CN118303578A (en) * | 2024-06-11 | 2024-07-09 | 南京西麦大健康科技有限公司 | Method for regulating and controlling GI value of oatmeal through ultrasonic wave synergistic granule-finishing solid-state enzymolysis |
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| CN115363100A (en) * | 2022-09-15 | 2022-11-22 | 草根知本集团有限公司 | Plant protein beverage rich in beta-glucan and preparation method thereof |
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