CN117903336A - Method for viscosity reduction alcohol-free extraction of high-purity beta-glucan by using oxidant - Google Patents
Method for viscosity reduction alcohol-free extraction of high-purity beta-glucan by using oxidant 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 relates to an extraction method of non-water-soluble beta-glucan, in particular to a method for viscosity reduction and alcohol-free extraction of high-purity beta-glucan (such as curdlan) alkali solution by using an oxidant. Adding alkali into crude curdlan product suspension to prepare thick alkali solution, adding an oxidant to reduce viscosity, filtering the alkali solution after viscosity reduction to remove impurities by using a ceramic membrane, adding acid into the clarified beta-glucan alkali solution after impurity removal to neutralize, desalting by using the ceramic membrane, concentrating and drying to obtain a high-purity beta-glucan product, wherein the dry purity of the product is more than 90% (w/w), and the extraction yield is more than 80% (w/w). Compared with the traditional extraction technology, the extraction and purification technology of the invention has the advantages that the content of the alkali-soluble curdlan is obviously improved, the technology is simple, the whole process is alcohol-free extraction, and the extraction method is safer. The concentration of glucan is more than or equal to 5% when the method is used for alkali dissolution, the method can obviously reduce the purification cost, and simultaneously solve the problem of non-gel unqualified product treatment of the stock of the curdlan production enterprises.
Description
Technical Field
The invention relates to a method for extracting beta-glucan, in particular to a method for viscosity-reducing alcohol-free extraction of high-purity beta-glucan (such as curdlan) alkali solution by using an oxidant.
Background
Beta-glucans are a class of polysaccharides whose main chain is linked by beta-1, 3-glycosidic linkages, often containing beta-1, 2/beta-1, 4/beta-1, 6-linked branches in varying proportions and sizes depending on the source.
The beta-glucan has the biological activities of enhancing immunity, resisting tumors, resisting oxidation, resisting bacteria, resisting viruses, resisting fungi, reducing cholesterol, reducing blood fat and the like, is a good biological effect regulator, and is widely applied to a plurality of fields of medicines, foods, cosmetics, animal feed additives and the like.
Curdlan is synthesized by bacteria of rhizobium genus, and is water-insoluble beta-glucan formed by connecting beta-1, 3-D-glycosidic bonds, and currently, products sold on the market of domestic enterprises are crude products containing thalli and impurities, wherein the glucan content of the crude products is only 78% -80%, and the Curdlan is used as a gel for the food industry, and the gel is mainly formed by heating the Curdlan.
The gel property of the curdlan is given by the triple helix molecular conformation of the special beta-glucan, the normal product is gel type, but the gel strength of the crude curdlan product is found to be reduced along with the storage time in production practice, the amplitude reduction is positively correlated with the time, and finally the curdlan product is naturally changed into a non-gel type product, the detection shows that the molecular weight of the non-gel type product is not changed, the research shows that the natural conversion of gel type into non-gel type is caused by the change of the molecular space conformation, namely the collapse of the original compact triple helix structure into a loose triple helix structure is a non-reversible process. Therefore, the non-gel unqualified products with the gel property lost cause certain stock backlog, the phase change increases the operation cost of enterprises, and the provision of a digestion application way for the enterprises becomes urgent.
The various derivatives of beta-glucan can be applied to high-added-value industries such as medicines, cosmetics and the like due to the unique biological activity of the derivatives. The non-gel product of the curdlan is also suitable for developing the beta-glucan derivative, but the existence of impurities such as thalli in a crude product seriously affects the preparation quality of the derivative and the separation of the process. The purification of crude curdlan products is a prerequisite for the application of curdlan non-gel products in the development of beta-glucan derivatives.
The original curdlan purification process aims to ensure the stability of the molecular weight and molecular structure of curdlan so as to maintain the gel strength of the product, and adopts a physical adsorption or mechanical viscosity reduction means with relatively mild conditions to remove impurities after alkali dissolution. The viscosity of the curdlan alkali solution rises rapidly with the increase of the curdlan content, so that the curdlan alkali solution subjected to physical adsorption or mechanical viscosity reduction treatment can be adopted, but the total curdlan content is lower: physical adsorption methods are often less than 2.5% (w/v), and mechanical viscosity reduction methods are also not more than 4% (w/v); the details are shown in table 1 below.
TABLE 1
Disclosure of Invention
The invention mainly aims to provide a method for extracting high-purity beta-glucan by using an oxidant to reduce viscosity without alcohol. The method not only reduces the purification cost, solves the problem of processing non-gel type unqualified products in stock of the curdlan production enterprises, reduces the operation cost, but also provides a new idea for carrying out the chemical modification of the beta-glucan.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
The invention provides a method for viscosity reduction and extraction of beta-glucan by using an oxidant, which comprises the following steps:
s1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution;
S2, adding an oxidant into the curdlan alkali solution, and stirring and reacting to prepare a low-viscosity beta-glucan alkali solution;
S3, separating the low-viscosity beta-glucan alkali solution by using a ceramic membrane to obtain a clarified beta-glucan alkali solution;
s4, adding an acid solution into the clarified beta-glucan alkali solution, neutralizing until the pH is neutral, and separating out beta-glucan neutralization gel to obtain a salt-containing beta-glucan neutralization gel aqueous suspension;
S5, desalting, concentrating and drying the salt-containing beta-glucan aqueous suspension by using a ceramic membrane to obtain the beta-glucan aqueous suspension.
Further, in the step S1, the concentration of beta-glucan in the dissolution liquid is controlled to be 5.0% -10.0% (w/v), and the concentration of hydroxyl ions is controlled to be 0.5-1.5 mol/L.
Further, the addition amount of the oxidant in the step S2 is 0.5-2.5% of the volume of the curdlan alkali solution, the mixture is stirred and reacted for 6-12 hours, and the temperature is controlled to be 25-50 ℃.
Further, the oxidant is one or more selected from sodium hypochlorite, hydrogen peroxide, sodium peroxide and ozone.
Further, in the step S3 and the step S5, ceramic membrane filtration is performed at a temperature of 25 to 40 ℃.
Further, the pore diameter of the ceramic membrane is 50-200 nm.
Further, in step S5, when the filtrate conductivity decreases to 500 to 1200. Mu.s/cm, desalting is stopped.
Further, in the step S5, the beta-glucan is concentrated until the mass concentration of the beta-glucan is more than or equal to 5 percent.
Further, in step S5, spray drying is adopted, the inlet temperature of the spray dryer is 110-140 ℃, and the outlet temperature is 60-90 ℃.
The invention also provides the beta-glucan prepared by the method.
Compared with the prior art, the invention has the following technical advantages:
The method comprises the steps of adding an oxidant into an alkali-soluble curdlan crude product for viscosity reduction, filtering the alkali solution after viscosity reduction by using a ceramic membrane to remove impurities, adding acid into the clarified beta-glucan alkali solution after impurity removal for neutralization, desalting by using the ceramic membrane, concentrating and drying to obtain a high-purity beta-glucan product, wherein the dry purity of the product is more than 90% (w/w), and the extraction yield is more than 80% (w/w).
Compared with the traditional extraction technology, the extraction and purification technology of the invention has the advantages that the content of the alkali-soluble curdlan is obviously improved, the technology is simple, the whole process is alcohol-free extraction, and the extraction method is safer. The concentration of glucan is more than or equal to 5% when the method is used for alkali dissolution, and the method can obviously reduce the purification cost.
Drawings
FIG. 1 is a flow chart of a method for viscosity reduction extraction of beta-glucan using an oxidizing agent in accordance with the present invention.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present invention. As used herein, the singular forms also are intended to include the plural forms unless the context clearly indicates otherwise, and furthermore, it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, and/or combinations thereof.
In order to enable those skilled in the art to more clearly understand the technical scheme of the present invention, the technical scheme of the present invention will be described in detail with reference to specific embodiments.
Example 1
A method for viscosity reduction extraction of beta-glucan by using an oxidant, which comprises the following steps:
S1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution; controlling the concentration of the curdlan in the dissolution liquid to be 5.0% (w/v), and the concentration of OH < - > to be 1.0mol/L; the alkali solution is sodium hydroxide solution.
S2, adding hydrogen peroxide into the curdlan alkali solution, wherein the addition amount of the oxidant is 0.5% of the volume of the curdlan alkali solution, stirring and reacting for 8 hours, and controlling the temperature to 40 ℃ to obtain the low-viscosity curdlan alkali solution.
S3, filtering and concentrating the obtained low-viscosity curdlan alkali solution by using a ceramic membrane, controlling the operation temperature to be 40 ℃, and recovering the filtered alkali solution to obtain a clarified beta-glucan alkali solution; the pore diameter of the ceramic membrane is 100nm.
S4, adding acetic acid solution into the clarified beta-glucan alkali solution, neutralizing to pH7.0, and separating out beta-glucan in the form of white gel to obtain beta-glucan high-salt-content gel; the concentration of the acetic acid solution is 1.0mol/L.
S5, filtering and concentrating the obtained beta-glucan gel with a ceramic membrane, controlling the operation temperature at 40 ℃, discharging the filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding purified water when the conductivity of the filtrate is reduced to 600 mu S/cm, and continuously concentrating until the mass concentration of the beta-glucan reaches 5% (w/v), thus obtaining beta-glucan aqueous suspension; the pore diameter of the ceramic membrane is 100nm.
S6, drying: drying the beta-glucan aqueous suspension by a spray dryer, wherein the inlet temperature of the spray dryer is 120 ℃, the outlet temperature of the spray dryer is 80 ℃, the beta-glucan powder is obtained, the purity of the product is over 90.8% (w/w), the glucan yield is 80.75%, and the non-gel beta-1, 3-glucan product is obtained after packaging.
Example 2
A method for viscosity reduction extraction of beta-glucan by using an oxidant, which comprises the following steps:
S1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution; controlling the concentration of the curdlan in the dissolution liquid to be 5.0% (w/v), and the concentration of OH < - > to be 1.0mol/L; the alkali solution is sodium hydroxide solution.
S2, adding hydrogen peroxide into the curdlan alkali solution, wherein the addition amount of the oxidant is 0.5% of the volume of the curdlan alkali solution, stirring and reacting for 8 hours, and controlling the temperature to 40 ℃ to obtain the low-viscosity curdlan alkali solution.
S3, filtering and concentrating the obtained low-viscosity curdlan alkali solution by using a ceramic membrane, controlling the operation temperature to be 40 ℃, and recovering the filtered alkali solution to obtain a clarified beta-glucan alkali solution; the pore diameter of the ceramic membrane is 50nm.
S4, neutralization precipitation: adding acetic acid solution into the clarified glucan alkali solution, neutralizing until the pH value is 7.0, and separating out beta-glucan in white gel to obtain beta-glucan high-salt-content gel; the concentration of the acetic acid solution is 1.0mol/L.
S5, filtering and concentrating the obtained beta-glucan gel with a ceramic membrane, controlling the operation temperature at 40 ℃, discharging the filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding purified water when the conductivity of the filtrate is reduced to 600 mu S/cm, and continuously concentrating until the final mass concentration of the beta-glucan reaches 5% (w/v), thus obtaining beta-glucan aqueous suspension; the pore diameter of the ceramic membrane is 50nm.
S6, drying: and (3) drying the beta-glucan aqueous suspension by a spray dryer, wherein the inlet temperature of the spray dryer is 120 ℃, the outlet temperature of the spray dryer is 80 ℃, the beta-glucan powder is obtained, the product purity is over 92.1% (w/w), the glucan yield is 82.32%, and the non-gel beta-1, 3-glucan product is obtained after packaging.
Example 3
A method for viscosity reduction extraction of beta-glucan by using an oxidant, which comprises the following steps:
S1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution; controlling the concentration of the curdlan in the dissolution liquid to be 5.0% (w/v), and the concentration of OH - to be 1.0mol/L; the alkali solution is sodium hydroxide solution.
S2, adding hydrogen peroxide into the curdlan alkali solution, wherein the addition amount of the oxidant is 0.5% of the volume of the curdlan alkali solution, stirring and reacting for 8 hours, and controlling the temperature to 40 ℃ to obtain the low-viscosity curdlan alkali solution.
S3, filtering and concentrating the obtained low-viscosity curdlan alkali solution by using a ceramic membrane, controlling the operation temperature to be 40 ℃, and recovering the filtered alkali solution to obtain a clarified beta-glucan alkali solution; the pore diameter of the ceramic membrane is 200nm.
S4, adding acetic acid solution into the clarified beta-glucan alkali solution, neutralizing to pH7.0, and separating out beta-glucan in the form of white gel to obtain beta-glucan high-salt-content gel; the concentration of the acetic acid solution is 1.0mol/L.
S5, filtering and concentrating the obtained beta-glucan gel with a ceramic membrane, controlling the operation temperature at 40 ℃, discharging the filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding purified water when the conductivity of the filtrate is reduced to 600 mu S/cm, and continuously concentrating until the mass concentration of the beta-glucan reaches 5% (w/v), thus obtaining beta-glucan aqueous suspension; the pore diameter of the ceramic membrane is 50nm.
S6, drying: and (3) drying the beta-glucan aqueous suspension by a spray dryer, wherein the inlet temperature of the spray dryer is 120 ℃, the outlet temperature of the spray dryer is 80 ℃, the beta-glucan powder is obtained, the purity of the product exceeds 93.2% (w/w), the glucan yield is 80.13%, and the non-gel beta-1, 3-glucan product is obtained after packaging.
Example 4
A method for viscosity reduction extraction of beta-glucan by using an oxidant, which comprises the following steps:
S1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution; controlling the concentration of the curdlan in the dissolution liquid to be 10.0% (w/v), and the concentration of OH - to be 1.5mol/L; the alkali solution is sodium hydroxide solution.
S2, adding sodium hypochlorite into the curdlan alkali solution, wherein the adding amount of the sodium hypochlorite is 2.5% of the volume of the curdlan alkali solution, stirring and reacting for 12 hours, and controlling the temperature at 50 ℃ to obtain the low-viscosity curdlan alkali solution.
S3, filtering and concentrating the obtained low-viscosity curdlan alkali solution by using a ceramic membrane, controlling the operation temperature to be 50 ℃, and recovering the filtered alkali solution to obtain a clarified beta-glucan alkali solution; the pore diameter of the ceramic membrane is 100nm.
S4, adding acetic acid solution into the clarified beta-glucan alkali solution, neutralizing to pH7.0, and separating out beta-glucan in the form of white gel to obtain beta-glucan high-salt-content gel; the concentration of the acetic acid solution is 1.0mol/L.
S5, filtering and concentrating the obtained beta-glucan gel with a ceramic membrane, controlling the operation temperature at 40 ℃, discharging the filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding purified water when the conductivity of the filtrate is reduced to 500 mu S/cm, and continuously concentrating until the mass concentration of the beta-glucan reaches 5% (w/v), thus obtaining beta-glucan aqueous suspension; the pore diameter of the ceramic membrane is 100nm.
S6, drying: drying the beta-glucan aqueous suspension by a spray dryer, wherein the inlet temperature of the spray dryer is 140 ℃, the outlet temperature of the spray dryer is 90 ℃, the beta-glucan powder is obtained, the product purity is over 93.8% (w/w), the glucan yield is 79.6%, and the non-gel beta-1, 3-glucan product is obtained after packaging.
Example 4
A method for viscosity reduction extraction of beta-glucan by using an oxidant, which comprises the following steps:
S1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution; controlling the concentration of the curdlan in the dissolution liquid to be 8.0% (w/v), and the concentration of OH - to be 0.5mol/L; the alkali solution is sodium hydroxide solution.
S2, adding sodium hypochlorite into the curdlan alkali solution, wherein the adding amount of the sodium hypochlorite is 1.5% of the volume of the curdlan alkali solution, stirring and reacting for 6 hours, and controlling the temperature to 25 ℃ to obtain the curdlan alkali solution with low viscosity.
S3, filtering and concentrating the obtained low-viscosity curdlan alkali solution by using a ceramic membrane, controlling the operation temperature to be 25 ℃, and recovering the filtered alkali solution to obtain a clarified beta-glucan alkali solution; the pore diameter of the ceramic membrane is 100nm.
S4, adding acetic acid solution into the clarified beta-glucan alkali solution, neutralizing to pH7.0, and separating out beta-glucan in the form of white gel to obtain beta-glucan high-salt-content gel; the concentration of the acetic acid solution is 1.0mol/L.
S5, filtering and concentrating the obtained beta-glucan gel with a ceramic membrane, controlling the operation temperature to be 25 ℃, discharging a filtrate, continuously adding purified water into the concentrated solution in the concentration process, stopping adding purified water when the conductivity of the filtrate is reduced to 1200 mu S/cm, and continuously concentrating until the mass concentration of the beta-glucan reaches 5% (w/v), so as to obtain a beta-glucan aqueous suspension; the pore diameter of the ceramic membrane is 100nm.
S6, drying: drying the beta-glucan aqueous suspension by a spray dryer, wherein the inlet temperature of the spray dryer is 110 ℃, the outlet temperature of the spray dryer is 60 ℃, the beta-glucan powder is obtained, the purity of the product is over 90.1% (w/w), the glucan yield is 78.6%, and the non-gel beta-1, 3-glucan product is obtained after packaging.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. A method for viscosity reduction alcohol-free extraction of high-purity beta-glucan by using an oxidant, which is characterized by comprising the following steps:
s1, preparing a crude curdlan product into a suspension, adding the suspension into an alkali solution, and continuously stirring to completely dissolve the suspension to obtain a high-viscosity curdlan alkali solution;
S2, adding an oxidant into the curdlan alkali solution, and stirring and reacting to prepare a low-viscosity beta-glucan alkali solution;
S3, separating the low-viscosity beta-glucan alkali solution by using a ceramic membrane to obtain a clarified beta-glucan alkali solution;
s4, adding an acid solution into the clarified beta-glucan alkali solution, neutralizing until the pH is neutral, and separating out beta-glucan in a gel form to obtain a salt-containing beta-glucan aqueous suspension;
S5, desalting, concentrating and drying the salt-containing beta-glucan aqueous suspension by using a ceramic membrane to obtain the beta-glucan aqueous suspension.
2. The method according to claim 1, wherein the concentration of β -glucan in the solution is controlled to be 5.0% to 10.0% (w/v) and the concentration of hydroxide ion is controlled to be 0.5 to 1.5mol/L in the step S1.
3. The method according to claim 1, wherein the oxidant is added in the amount of 0.5-2.5% of the volume of the curdlan alkali solution in the step S2, and the reaction is carried out for 6-12 hours under stirring, and the temperature is controlled to be 25-50 ℃.
4. A method according to claim 1 or 3, wherein the oxidizing agent is selected from one or more of sodium hypochlorite, hydrogen peroxide, sodium peroxide, ozone.
5. The method according to claim 1, wherein the ceramic membrane filtration is performed at a temperature of 25 to 40 ℃ in step S3 and step S5.
6. The method according to claim 1 or 5, wherein the ceramic membrane has a pore size of 50 to 200nm.
7. The method according to claim 1, wherein in step S5, the desalting is stopped when the filtrate conductivity falls to 500 to 1200 μs/cm.
8. The method according to claim 1, wherein in step S5, the concentration is carried out to a concentration of beta-glucan of 5% or more by mass.
9. The method according to claim 1, wherein in step S5, spray drying is used, the spray dryer having an inlet temperature of 110 to 140 ℃ and an outlet temperature of 60 to 90 ℃.
10. The beta-glucan produced by the process of any one of claims 1-9.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103897073A (en) * | 2014-03-12 | 2014-07-02 | 山东省食品发酵工业研究设计院 | Method for separating and purifying curdlan by use of flocculant |
| CN103897074A (en) * | 2014-03-12 | 2014-07-02 | 山东省食品发酵工业研究设计院 | Method for separating and purifying curdlan by use of membrane process |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103897073A (en) * | 2014-03-12 | 2014-07-02 | 山东省食品发酵工业研究设计院 | Method for separating and purifying curdlan by use of flocculant |
| CN103897074A (en) * | 2014-03-12 | 2014-07-02 | 山东省食品发酵工业研究设计院 | Method for separating and purifying curdlan by use of membrane process |
Non-Patent Citations (1)
| Title |
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| 王英杰: "低分子量水溶性β-1，3-葡聚糖的制备及其活性研究", 中国优秀硕士学位论文全文数据库工程科技Ⅰ辑, no. 8, 15 August 2014 (2014-08-15), pages 014 - 304 * |
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