CN112794927A - Method for extracting sea cucumber polysaccharide with high purity and high extraction rate - Google Patents
Method for extracting sea cucumber polysaccharide with high purity and high extraction rate Download PDFInfo
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- CN112794927A CN112794927A CN202110265213.7A CN202110265213A CN112794927A CN 112794927 A CN112794927 A CN 112794927A CN 202110265213 A CN202110265213 A CN 202110265213A CN 112794927 A CN112794927 A CN 112794927A
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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Abstract
The invention discloses a method for extracting sea cucumber polysaccharide with high purity and high extraction rate, and relates to the field of polysaccharide extraction. Extracting by using a mode of combining enzymolysis and leaching, wherein an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent in the enzymolysis process of the sea cucumber, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is (1-100): 1000. the novel process method realizes the preservation of the maximum active substances of the sea cucumber polysaccharide, solves the problem that the traditional process can not consider both high purity and high extraction rate, realizes the preservation of the maximum active substances of the sea cucumber polysaccharide, has simple extraction and purification process, can meet the requirements of products, and realizes the effects of high purity and high extraction rate of the sea cucumber polysaccharide.
Description
Technical Field
The invention relates to the field of polysaccharide extraction, in particular to a method for extracting sea cucumber polysaccharide with high purity and high extraction rate.
Background
The study of polysaccharides, the first of which was in the 40's of the 20 th century, where the earlier and most studied, was various capsular polysaccharides obtained from bacteria, which were used medically mainly for vaccines. However, polysaccharides have attracted considerable attention as broad-spectrum immunopotentiators in the 60's of the 20 th century. In particular, in the last two decades, due to the development of molecular biology, it has been gradually recognized that saccharides and complex molecules thereof have important biological functions, and a special conference about "glycobiology and glycoengineering" was called many times in succession, and in 1984, the twelfth international carbohydrate conference called in the netherlands by the soviet union reported that capsular polysaccharide with a specific structure is synthesized completely as a vaccine, and the capsular polysaccharide has received great interest from participants.
The difficulty of the traditional sea cucumber extraction process is that when the sea cucumber polysaccharide is extracted, the enzymolysis process adopts a novel extraction method combining enzyme and extraction, and utilizes the catalytic activity and selectivity of the enzyme to destroy the cell wall structure under mild conditions to promote the dissolution and release of intracellular components so as to achieve the extraction purpose. When the extraction rate is improved, the purity of the sea cucumber polysaccharide is difficult to ensure, and when the purity of the sea cucumber polysaccharide is ensured, the extraction rate is lower, and the separation and purification technology of the sea cucumber polysaccharide is still more difficult at present.
Disclosure of Invention
The invention aims to provide a method for extracting sea cucumber polysaccharide with high purity and high extraction rate, which realizes the preservation of the maximum active substances of the processed sea cucumber polysaccharide by a novel process method, solves the problem that the traditional process can not realize both high purity and high extraction rate, realizes the preservation of the maximum active substances of the processed sea cucumber polysaccharide, has simple extraction and purification process, can meet the requirements of products, and realizes the effects of high purity and high extraction rate of the sea cucumber polysaccharide.
In order to achieve the effects, the application discloses a method for extracting sea cucumber polysaccharide with high purity and high extraction rate, wherein enzymolysis and extraction are combined for extraction, an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent in the enzymolysis process of the sea cucumber, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is (1-100): 1000.
further, the organic acid in the retention accelerator is selected from citric acid, malic acid, and ethylenediaminetetraacetic acid, and trisodium citrate is exemplified as a salt of citric acid. As the salt of malic acid, disodium malate may be mentioned. Examples of salts of ethylenediaminetetraacetic acid include disodium ethylenediaminetetraacetate and trisodium ethylenediaminetetraacetate.
Further wherein the step of enzymatic hydrolysis uses a protease for enzymatic hydrolysis.
Further wherein the leaching process uses ultrasonic dispersive temperature rise for leaching.
Further, the method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure at the temperature of 100 ℃ and the pressure of 0.1-0.3MPa until the sea cucumber is soft;
(2) crushing, namely adding 5-10 times of water by mass into the cured sea cucumber, and stirring at a high speed for crushing. Obtaining sea cucumber pulp;
(3) performing enzymolysis, namely adding a promoting and maintaining agent into the sea cucumber pulp, and adding a proper amount of protease for enzymolysis;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
Further, the protease is selected from compound protease mixed by papain and neutral protease according to the ratio of 1: 1.
The invention has the beneficial effects that:
the method has the advantages that the certain amount of the promoting and maintaining agent is added in the enzymolysis process, the enzymolysis process can be promoted, the yield of the polysaccharide after enzymolysis can be improved, the purity of the polysaccharide is improved to a certain extent, and the method can realize polysaccharide extraction on the sea cucumber with high purity and high extraction rate by matching the promoting and maintaining agent with enzymolysis and extraction. Solves the problem of the traditional process.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all 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.
The first embodiment is as follows:
a method for extracting sea cucumber polysaccharide with high purity and high extraction rate adopts a mode of combining enzymolysis and extraction for extraction, and is characterized in that: in the enzymolysis process of the sea cucumber, an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is 25: 1000.
wherein the organic acid in the retention aid is selected from the group consisting of citric acid, malic acid, trisodium citrate and disodium malate. Further wherein the step of enzymatic hydrolysis uses a protease for enzymatic hydrolysis.
Wherein the leaching process uses ultrasonic dispersion to increase the temperature for leaching. The method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure, wherein the temperature is 120 ℃, and the pressure is 0.2MPa until the sea cucumber is soft;
(2) crushing, namely adding 5 times of water by mass into the cured sea cucumber, and stirring at a high speed for crushing. Obtaining sea cucumber pulp;
(3) performing enzymolysis, namely adding a promoting and maintaining agent into the sea cucumber pulp, and adding a proper amount of protease for enzymolysis, wherein the protease is selected from compound protease formed by mixing papain and neutral protease according to the ratio of 1: 1;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
Example two:
a method for extracting sea cucumber polysaccharide with high purity and high extraction rate adopts a mode of combining enzymolysis and extraction for extraction, and is characterized in that: in the enzymolysis process of the sea cucumber, an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is 50: 1000.
wherein the organic acid in the retention aid is selected from the group consisting of citric acid, malic acid, trisodium citrate and disodium malate. Further wherein the step of enzymatic hydrolysis uses a protease for enzymatic hydrolysis.
Wherein the leaching process uses ultrasonic dispersion to increase the temperature for leaching. The method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure, wherein the temperature is 120 ℃, and the pressure is 0.2MPa until the sea cucumber is soft;
(2) crushing, namely adding 5 times of water by mass into the cured sea cucumber, and stirring at a high speed for crushing. Obtaining sea cucumber pulp;
(3) performing enzymolysis, namely adding a promoting and maintaining agent into the sea cucumber pulp, and adding a proper amount of protease for enzymolysis, wherein the protease is selected from compound protease formed by mixing papain and neutral protease according to the ratio of 1: 1;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
Example three:
a method for extracting sea cucumber polysaccharide with high purity and high extraction rate adopts a mode of combining enzymolysis and extraction for extraction, and is characterized in that: in the enzymolysis process of the sea cucumber, an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is 100: 1000.
wherein the organic acid in the retention aid is selected from the group consisting of citric acid, malic acid, trisodium citrate and disodium malate. Further wherein the step of enzymatic hydrolysis uses a protease for enzymatic hydrolysis.
Wherein the leaching process uses ultrasonic dispersion to increase the temperature for leaching. The method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure, wherein the temperature is 120 ℃, and the pressure is 0.2MPa until the sea cucumber is soft;
(2) crushing, namely adding 5 times of water by mass into the cured sea cucumber, and stirring at a high speed for crushing. Obtaining sea cucumber pulp;
(3) performing enzymolysis, namely adding a promoting and maintaining agent into the sea cucumber pulp, and adding a proper amount of protease for enzymolysis, wherein the protease is selected from compound protease formed by mixing papain and neutral protease according to the ratio of 1: 1;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
Comparative example one:
a method for extracting high-purity and high-extraction-rate Stichopus japonicus polysaccharide comprises extracting Stichopus japonicus polysaccharide by enzymolysis and extraction,
wherein the leaching process uses ultrasonic dispersion to increase the temperature for leaching. The method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure, wherein the temperature is 120 ℃, and the pressure is 0.2MPa until the sea cucumber is soft;
(2) crushing, namely adding 5 times of water by mass into the cured sea cucumber, and stirring at a high speed for crushing. Obtaining sea cucumber pulp;
(3) performing enzymolysis, namely adding a proper amount of protease into the sea cucumber pulp for enzymolysis, wherein the protease is selected from compound protease formed by mixing papain and neutral protease according to the ratio of 1: 1;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
Comparative example two:
a method for extracting sea cucumber polysaccharide with high purity and high extraction rate adopts a mode of combining enzymolysis and extraction for extraction, and is characterized in that: in the enzymolysis process of the sea cucumber, an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is 200: 1000.
wherein the organic acid in the retention aid is selected from the group consisting of citric acid, malic acid, trisodium citrate and disodium malate. Further wherein the step of enzymatic hydrolysis uses a protease for enzymatic hydrolysis.
Wherein the leaching process uses ultrasonic dispersion to increase the temperature for leaching. The method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure, wherein the temperature is 120 ℃, and the pressure is 0.2MPa until the sea cucumber is soft;
(2) crushing, namely adding 5 times of water by mass into the cured sea cucumber, and stirring at a high speed for crushing. Obtaining sea cucumber pulp;
(3) performing enzymolysis, namely adding a promoting and maintaining agent into the sea cucumber pulp, and adding a proper amount of protease for enzymolysis, wherein the protease is selected from compound protease formed by mixing papain and neutral protease according to the ratio of 1: 1;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
Experimental detection
The principle of the method is that the polysaccharide is firstly hydrolyzed into monosaccharide molecules under the action of sulfuric acid, and is quickly dehydrated to generate furfural derivatives, then the furfural derivatives and phenol are subjected to color reaction, and the absorbance and the sugar content are in a linear relationship at 490nm, so that the polysaccharide in a sample can be measured.
Purity detection glucose was used as a standard for sugar content testing as an index for evaluation of polysaccharide purity.
As can be seen from the data, according to the comparison of the first example, the second example, the third example and the first comparative example, the promotion and maintenance agent can obviously improve the purity and the extraction rate of the polysaccharide extraction, and the second comparative example can see that the promotion and maintenance agent in more excess is not beneficial to improving the extraction effect.
The above-mentioned embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solution of the present invention by those skilled in the art without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.
Claims (6)
1. A method for extracting sea cucumber polysaccharide with high purity and high extraction rate adopts a mode of combining enzymolysis and extraction for extraction, and is characterized in that: in the enzymolysis process of the sea cucumber, an aqueous solution containing at least one component of organic acid and salt thereof is used as a promoting and maintaining agent, and the adding proportion of the weight of solid matters in the promoting and maintaining agent to the weight of the sea cucumber raw material is (1-100): 1000.
2. the method for extracting sea cucumber polysaccharide with high purity and high extraction rate as claimed in claim 1, wherein the method comprises the following steps: the organic acid in the agent is selected from citric acid, malic acid, and ethylenediaminetetraacetic acid, and as a salt of citric acid, trisodium citrate; as the salt of malic acid, disodium malate; examples of salts of ethylenediaminetetraacetic acid include disodium ethylenediaminetetraacetate and trisodium ethylenediaminetetraacetate.
3. The method for extracting sea cucumber polysaccharide with high purity and high extraction rate as claimed in claim 1, wherein the method comprises the following steps: the step of enzymolysis uses protease for enzymolysis.
4. The method for extracting sea cucumber polysaccharide with high purity and high extraction rate as claimed in claim 1, wherein the method comprises the following steps: the leaching process uses ultrasonic dispersion and temperature rise for leaching.
5. The method for extracting sea cucumber polysaccharide with high purity and high extraction rate as claimed in claim 1, wherein the method comprises the following steps: the method specifically comprises the following steps:
(1) curing the sea cucumber, namely curing the raw sea cucumber at high temperature and high pressure at the temperature of 100 ℃ and the pressure of 0.1-0.3MPa until the sea cucumber is soft;
(2) crushing, namely adding 5-10 times of water by mass into the cured sea cucumber, and stirring and crushing at a high speed to obtain sea cucumber pulp;
(3) performing enzymolysis, namely adding a promoting and maintaining agent into the sea cucumber pulp, and adding a proper amount of protease for enzymolysis;
(4) leaching, using ultrasonic waves to promote leaching effect, adjusting ph to precipitate protein and amino acid, and reserving supernatant;
(5) rough preparation and purification, alcohol precipitation and impurity removal, and then ultrafiltration treatment is carried out to carry out rough preparation and purification on polysaccharide;
(6) freeze drying to obtain the final product.
6. The method for extracting sea cucumber polysaccharide with high purity and high extraction rate as claimed in claim 5, wherein the method comprises the following steps: the protease is selected from compound protease mixed by papain and neutral protease according to the proportion of 1: 1.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1749283A (en) * | 2005-10-11 | 2006-03-22 | 大连轻工业学院 | Mussel polysacharide and its preparing method |
| CN101248890A (en) * | 2008-04-03 | 2008-08-27 | 侯书胜 | Nutrition reinforced type high activity beche-de-mer products and method of preparing the same |
| CN109512696A (en) * | 2018-12-18 | 2019-03-26 | 海南热带海洋学院 | A kind of skin care item and preparation method thereof containing complex polysaccharide series |
| CN111393537A (en) * | 2020-04-08 | 2020-07-10 | 潍坊悦龙堂海洋科技有限公司 | Method for extracting bioactive substances from sea cucumber |
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- 2021-03-11 CN CN202110265213.7A patent/CN112794927A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1749283A (en) * | 2005-10-11 | 2006-03-22 | 大连轻工业学院 | Mussel polysacharide and its preparing method |
| CN101248890A (en) * | 2008-04-03 | 2008-08-27 | 侯书胜 | Nutrition reinforced type high activity beche-de-mer products and method of preparing the same |
| CN109512696A (en) * | 2018-12-18 | 2019-03-26 | 海南热带海洋学院 | A kind of skin care item and preparation method thereof containing complex polysaccharide series |
| CN111393537A (en) * | 2020-04-08 | 2020-07-10 | 潍坊悦龙堂海洋科技有限公司 | Method for extracting bioactive substances from sea cucumber |
Non-Patent Citations (1)
| Title |
|---|
| 穆琳: "加工对海参营养和功能成分", 《中国优秀博硕士学位论文全文数据库（硕士）工程科技I辑》 * |
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Application publication date: 20210514 |