CN101020914A - Acid and conjugated enzyme process for preparing fucan in lower molecular weight - Google Patents
Acid and conjugated enzyme process for preparing fucan in lower molecular weight Download PDFInfo
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- CN101020914A CN101020914A CNA2007100086731A CN200710008673A CN101020914A CN 101020914 A CN101020914 A CN 101020914A CN A2007100086731 A CNA2007100086731 A CN A2007100086731A CN 200710008673 A CN200710008673 A CN 200710008673A CN 101020914 A CN101020914 A CN 101020914A
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Abstract
The present invention discloses acid and conjugated enzyme process for preparing fucan in lower molecular weight. Coarse fucan is enzymolyzed with commercial pectinase under controlled enzymolysis time, temperature and pH value and intercepted with ultrafiltering film of molecular weight 1000-5000 Da to obtain low molecular weight fucan. The present invention has simple operation, mild condition, high yield, low sulfate radical dropping rate, low cost and other advantages, and is easy use in industrial production. The present invention has simultaneous fucan extraction and lower molecular weight fucan preparation, and has simplified technological process, high production efficiency and over 30 % lowered production cost. In addition, the process uses no harmful chemical reagent, and the product is safe, non-toxic and especially suitable for producing functional food and medicine.
Description
Technical field
The present invention relates to a kind of extracting method of fucosan, particularly relate to the method for utilizing acid system desmoenzyme legal system to be equipped with the lower molecular weight fucosan.
Background technology
Fucosan (Fucoidan) is present in and has higher bioactive crude substance in the marine alga, has antitumor, antiviral, anticoagulation, hypoglycemic blood fat, regulates immunizing power, removes effects such as active oxygen radical and inside and outside be anti-oxidant.Fucosan is the water-soluble polysaccharide that contains multiple monose and sulfate, and molecular mass does not wait from several ten thousand to hundreds of thousands of.Its special 26S Proteasome Structure and Function has caused many experts and scholars' attention.Along with deepening continuously of polysaccharide antivirus action mechanism research, the factor that discovery influences active polysaccharide is a lot, as content of the polymerization degree, molecular mass and the sulfate radical of the combining form of glycosidic link between link position, branch's density and the monose of the higher structure of polysaccharide, side chain, polysaccharide etc., all relevant with pharmacologically active.
The molecular weight that studies show that sulfated polysaccharide is at 1000-50000Da, and sulfate is higher 15～20% o'clock biological activitys.So will when reducing the sulfate amount of coming off, reduce the molecular weight of fucosan, be fucosan and products thereof exploitation problem anxious to be solved by appropriate means.At present the degradation method of the fucosan of report mainly contains two kinds of acidolysis and enzymolysis.Wherein strong acid hydrolysis reaction condition is violent, is difficult to operation, and product need carry out the desalination operation; And narrow spectrum Sargassum polysaccharides sulfuric ester lytic enzyme costs an arm and a leg, and output is little.
Summary of the invention
The object of the present invention is to provide a kind of simple to operate, mild condition, output height, the sulfate radical expulsion rate is low, cost is low acid system desmoenzyme legal system to be equipped with the method for lower molecular weight fucosan, this method can be applicable to suitability for industrialized production.
For achieving the above object, technical solution of the present invention is:
The present invention is the method that a kind of acid system desmoenzyme legal system is equipped with the lower molecular weight fucosan, it may further comprise the steps: (1) is 1 in the ratio of dry seaweed and acid: the ratio of 8-10 adds acid, extract after 2-4 hour, centrifugal collection supernatant liquor, add ethanolic soln to the 30%-40% saturation ratio, collect supernatant liquor and add ethanolic soln again, stir, leave standstill 20 minutes precipitations to the 70%-80% saturation ratio, the solution centrifugal collecting precipitation is got fucosan, lyophilize; (2) fucosan that extracted utilizes proteolytic enzyme time to remove albumen 37 ℃-45 ℃ and pH=2～3; (3) add polygalacturonase under 30 ℃ of-45 ℃ of temperature heating hydrolysis 2-3 hour; Being adjusted to the pH value with acid is 4-5, places 2-4 hour, filters or the centrifugal impurity of removing, and gets supernatant liquor; (4) selecting molecular weight cut-off is that the film of 1000-5000Da carries out ultrafiltration and concentration, the solution that concentrated is loaded in the beaker, pre-freeze 1-2 hour, after put into vacuum drying oven, cover lid mixes up vacuum tightness and temperature, and temperature is-55 ℃, about lyophilize one day, promptly obtaining molecular weight is the lower molecular weight fucosan of 1000-5000D.
Add acid in the step (1) and be 0.1-0.2N hydrochloric acid.
Adding proteolytic enzyme in the step (2) is stomach en-.
Enzyme described in the step (3) is a polygalacturonase.
After adopting such scheme, the present invention has adopted the thick fucosan of coml toolenzyme polygalacturonase hydrolysis, has controlled enzymolysis time, temperature and pH value, and the employing molecular weight cut-off is the ultra-filtration membrane of 1000-5000Da, obtains low-molecular-weight fucosan.Have simple to operate, mild condition, output height, low, the low cost and other advantages of sulfate radical expulsion rate, be easy to suitability for industrialized production.
Because the present invention adopts acid system directly to extract low-molecular-weight fucosan in conjunction with enzyme process from marine alga, the extraction of fucosan and the preparation of lower molecular weight fucosan can be carried out simultaneously, simplify technical process, improved production efficiency, saved productive expense more than 30%.Meanwhile, whole technology avoids using poisonous and hazardous chemical reagent, and the product safety of preparation is nontoxic, is particularly suitable for being applied to the production of functional food and medicine.
The fucosan of the present invention preparation has wide prospect in medicine, can be applicable to treat hepatitis B, antiviral, anticoagulation, hypoglycemic blood fat, regulates immunizing power, removes active oxygen radical and the oxidation resistant marine drug in inside and outside.
The present invention is further illustrated below in conjunction with specific embodiment.
Embodiment
Embodiment 1:
A kind of acid system desmoenzyme legal system is equipped with the method for lower molecular weight fucosan, may further comprise the steps:
(1) gets dried sea-tangle 1000kg and add acid in 1: 9 ratio, extract after 2 hours, centrifugal collection supernatant liquor adds ethanolic soln, to 30% saturation ratio, the collection supernatant liquor adds ethanolic soln to 70% saturation ratio again and stirs, leaves standstill 20 minutes precipitations, the solution centrifugal collecting precipitation is got fucosan, lyophilize.
(2) it is that the film of 1000Da carries out ultrafiltration that the fucosan that extracted is selected molecular weight cut-off, and utilize proteolytic enzyme 37 ℃ and pH=2～3 time except that albumen.Obtain the thick fucosan product of 21g.Get fucosan crude product 1g, utilize proteolytic enzyme time to remove albumen 37 ℃ and pH=2～3.
(3) add polygalacturonase heating hydrolysis 2 hours under 30 ℃ of temperature; Being adjusted to pH value with acid is 4, places 2 hours, filters or the centrifugal impurity of removing, and gets supernatant liquor.
(4) selecting molecular weight cut-off is that polysaccharide solution after the membrane ultrafiltration of 5000Da concentrates is loaded in the beaker, pre-freeze 1 hour, after put into vacuum drying oven, cover lid, mix up vacuum tightness and temperature, temperature is-55 ℃, dry about one day, uses freeze-drying to remove moisture and obtains lower molecular weight fucosan 0.70g.
Embodiment 2:
A kind of acid system desmoenzyme legal system is equipped with the method for lower molecular weight fucosan, may further comprise the steps:
(1) gets dried sea-tangle 1000 and add acid in 1: 10 ratio, extract after 4 hours, centrifugal collection supernatant liquor adds ethanolic soln, to 40% saturation ratio, the collection supernatant liquor adds ethanolic soln to 80% saturation ratio again and stirs, leaves standstill 20 minutes precipitations, the solution centrifugal collecting precipitation is got fucosan, lyophilize.
(2) it is that the film of 5000Da carries out ultrafiltration that the fucosan that extracted is selected molecular weight cut-off, and utilizes proteolytic enzyme to remove albumen under 45 ℃ and pH=2, obtains the thick fucosan product of 21g.Get fucosan crude product 1g, utilize proteolytic enzyme under 45 ℃ and pH=2, to remove albumen.
(3) add polygalacturonase heating hydrolysis 3 hours under 45 ℃ of temperature; Being adjusted to pH value with acid is 5, places 3 hours, filters or the centrifugal impurity of removing, and gets supernatant liquor.
(4) selecting molecular weight cut-off is that polysaccharide solution after the membrane ultrafiltration of 5000Da concentrates is loaded in the beaker, pre-freeze 2 hours, after put into vacuum drying oven, cover lid, mix up vacuum tightness and temperature, temperature is-55 ℃, dry about one day, uses freeze-drying to remove moisture and obtains the lower molecular weight fucosan.
Embodiment 3:
(1) gets dried sea-tangle 1000kg and add acid in 1: 8 ratio, extract after 3 hours, centrifugal collection supernatant liquor adds ethanolic soln, to 35% saturation ratio, the collection supernatant liquor adds ethanolic soln to 75% saturation ratio again and stirs, leaves standstill 20 minutes precipitations, the solution centrifugal collecting precipitation is got fucosan, lyophilize.
(2) it is that the film of 1000Da carries out ultrafiltration that the fucosan that extracted is selected molecular weight cut-off, and utilizes enzyme to remove albumen under 40 ℃ and pH=2.5, obtains thick fucosan product.Get the fucosan crude product, utilize proteolytic enzyme under 40 ℃ and pH=2.5, to remove albumen.
(3) add polygalacturonase heating hydrolysis 2.5 hours under 35 ℃ of temperature; Being adjusted to pH value with acid is 4.5, places 2.5 hours, filters or the centrifugal impurity of removing, and gets supernatant liquor.
(4) selecting molecular weight cut-off is that polysaccharide solution after the membrane ultrafiltration of 5000Da concentrates is loaded in the beaker, pre-freeze 1.5 hours, after put into vacuum drying oven, cover lid, mix up vacuum tightness and temperature, temperature is-55 ℃, dry about one day, uses freeze-drying to remove moisture and obtains the lower molecular weight fucosan.
Polygalacturonase of the present invention is commercially available polygalacturonase (Pectinase): pH4.0,40 ℃, 〉=1.0u/mg).
The used acid of the present invention is 0.1-0.2N hydrochloric acid.Used proteolytic enzyme is stomach en-.
The present invention extracts the lower molecular weight fucosan product of preparation from marine alga, check through authoritative department, the content of lower molecular weight fucosan reaches more than 98%, and adopting the barium sulfate weighting method to record sulfate content is 20.83%, and keeps original natural structure and biological activity.
Claims (4)
1, a kind of acid system desmoenzyme legal system is equipped with the method for lower molecular weight fucosan, it is characterized in that: it may further comprise the steps: (1) is 1 in the ratio of dry seaweed and acid: the ratio of 8-10 adds acid, extract after 2-4 hour, centrifugal collection supernatant liquor, add ethanolic soln to the 30%-40% saturation ratio, collect supernatant liquor and add ethanolic soln again, stir, leave standstill 20 minutes precipitations to the 70%-80% saturation ratio, the solution centrifugal collecting precipitation is got fucosan, lyophilize; (2) fucosan that extracted utilizes proteolytic enzyme time to remove albumen 37 ℃-45 ℃ and pH=2～3; (3) add polygalacturonase under 30 ℃ of-45 ℃ of temperature heating hydrolysis 2-3 hour; Being adjusted to the pH value with acid is 4-5, places 2-4 hour, filters or the centrifugal impurity of removing, and gets supernatant liquor; (4) selecting molecular weight cut-off is that the film of 1000-5000Da carries out ultrafiltration and concentration, the solution that concentrated is loaded in the beaker, pre-freeze 1-2 hour, after put into vacuum drying oven, cover lid mixes up vacuum tightness and temperature, and temperature is-55 ℃, about lyophilize one day, promptly obtaining molecular weight is the lower molecular weight fucosan of 1000-5000D.
2, preparation method according to claim 1 is characterized in that: add acid in the step (1) and be 0.1-0.2N hydrochloric acid.
3, preparation method according to claim 1 is characterized in that: adding proteolytic enzyme in the step (2) is stomach en-.
4, the method for utilizing marine alga to make the lower molecular weight fucosan according to claim 1, it is characterized in that: the enzyme described in the step (3) is a polygalacturonase.
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| Application Number | Priority Date | Filing Date | Title |
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| CN200710008673A CN100575499C (en) | 2007-03-07 | 2007-03-07 | Acid system desmoenzyme legal system is equipped with the method for lower molecular weight fucosan |
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| CN200710008673A CN100575499C (en) | 2007-03-07 | 2007-03-07 | Acid system desmoenzyme legal system is equipped with the method for lower molecular weight fucosan |
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| CN101020914A true CN101020914A (en) | 2007-08-22 |
| CN100575499C CN100575499C (en) | 2009-12-30 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659709B (en) * | 2009-09-18 | 2011-09-07 | 集美大学 | Preparation method of fucosan |
| CN102217768A (en) * | 2011-05-05 | 2011-10-19 | 集美大学 | Method for producing alga nutrient supplement |
| CN103338780A (en) * | 2010-12-20 | 2013-10-02 | 韩国食品研究院 | Composition for positively allosterically modulating GABAA-benzodiazepine receptor and composition for producing sedative-hypnotic effects containing ecklonia cava extract and ecklonia kurome extract |
| CN103554293A (en) * | 2013-11-18 | 2014-02-05 | 集美大学 | Preparation method and use of active low molecular weight fucosan |
| CN110272887A (en) * | 2019-07-05 | 2019-09-24 | 集美大学 | A kind of low-viscosity fucosan and its enzyme-linked coupling preparation of high temperature and pressure- |
-
2007
- 2007-03-07 CN CN200710008673A patent/CN100575499C/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101659709B (en) * | 2009-09-18 | 2011-09-07 | 集美大学 | Preparation method of fucosan |
| CN103338780A (en) * | 2010-12-20 | 2013-10-02 | 韩国食品研究院 | Composition for positively allosterically modulating GABAA-benzodiazepine receptor and composition for producing sedative-hypnotic effects containing ecklonia cava extract and ecklonia kurome extract |
| CN102217768A (en) * | 2011-05-05 | 2011-10-19 | 集美大学 | Method for producing alga nutrient supplement |
| CN103554293A (en) * | 2013-11-18 | 2014-02-05 | 集美大学 | Preparation method and use of active low molecular weight fucosan |
| CN103554293B (en) * | 2013-11-18 | 2016-08-17 | 集美大学 | A kind of preparation method and its usage of active low-molecular amount fucosan |
| CN110272887A (en) * | 2019-07-05 | 2019-09-24 | 集美大学 | A kind of low-viscosity fucosan and its enzyme-linked coupling preparation of high temperature and pressure- |
| CN110272887B (en) * | 2019-07-05 | 2020-12-04 | 集美大学 | Low-viscosity fucosan and high-temperature high-pressure enzyme-linked coupling preparation method thereof |
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| Publication number | Publication date |
|---|---|
| CN100575499C (en) | 2009-12-30 |
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Effective date of registration: 20100428 Address after: 361000 No. 3 Xiang Ming Road, Torch Industrial Zone (Xiangan), Fujian, Xiamen Patentee after: Xiamen City Sinong Food Co., Ltd. Address before: Yinjiang road in Jimei District of Xiamen City, Fujian Province, No. 185 361021 Patentee before: Jimei University |
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Address after: 361000 No. 3 Xiang Ming Road, Torch Industrial Zone (Xiangan), Fujian, Xiamen Patentee after: Xiamen Yanzhiwu Silken Food Co.,Ltd. Address before: 361000 No. 3 Xiang Ming Road, Torch Industrial Zone (Xiangan), Fujian, Xiamen Patentee before: XIAMEN SINONG FOOD Co.,Ltd. |
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