CN103936875A - Method for extracting laminarin from kelp - Google Patents
Method for extracting laminarin from kelp Download PDFInfo
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- CN103936875A CN103936875A CN201410156584.1A CN201410156584A CN103936875A CN 103936875 A CN103936875 A CN 103936875A CN 201410156584 A CN201410156584 A CN 201410156584A CN 103936875 A CN103936875 A CN 103936875A
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Abstract
The invention discloses a method for extracting laminarin from kelp. The method comprises the following steps: raw material treatment, ultrasonic treatment, enzymolysis, destaining, deproteinization and drying to obtain the product. Compared with the existing trichloroacetic acid precipitation method, savage method and the like, the method has the advantages that laminarin is high in yield and high in content.
Description
Technical field
The present invention relates to a kind of preparation method of polysaccharide, be specifically related to a kind of method of extracting laminarin from sea-tangle.
Background technology
Sea-tangle (Laminaria Japonica Aresch) is the kelp of phaeophyta Laminariales Laminariaceae Laminaria.Kelp Rich is containing several functions composition, have high edible and pharmaceutical use, its pharmaceutical use is all on the books in traditional medical books such as < < Compendium of Materia Medica > >, < < Shennong Bencaojing > > and < < food book on Chinese herbal medicine > >.Research shows that the multiple biological function of sea-tangle is mostly relevant with the physiologically active polysaccharide in sea-tangle, as immunomodulatory, antitumor, anticoagulation, lipopenicillinase, hypoglycemic, anti-oxidant etc.
At present, extracting method for laminarin has document or the report that some laminarins extract, traditional method is mainly acid extraction method, alkali extraction method, enzyme extraction method etc., all there is certain defect in these extracting method: acid is carried and can be caused polysaccharide hydrolysis, and alkali extraction method causes polysaccharide biological activity to reduce, and enzyme extraction method is consuming time, the enzyme amount that industrial production need to be used is large, cost is high, and the method for hot water extraction alcohol precipitation is easy, but time consumption and energy consumption.The method of albumen in polysaccharide that removes has trichloroacetic acid precipitation method and savage method etc., utilizes trichloroacetic acid precipitation albumen, and polysaccharide yield is higher, but often with sugar chain degradation; Savage method adopts the organic reagents such as chloroform and amylalcohol to make protein denaturation, precipitation, and its treatment condition are gentle, can avoid preferably polysaccharide degraded, but processing efficiency is low, and polysaccharide loss is serious.As Chinese patent CN103421126A, a kind of method of extracting laminarin is disclosed, after sea-tangle is cleaned, after drying, pulverize, sieving, be dipped in water, through water extraction and ultrasonic-leaching, and then alcohol precipitation, obtain laminarin precipitation, finally will precipitate oven dry, the laminarin of crude product, it is low that the method obtains laminarin content.The equal Shortcomings of existing method, this has restricted intensive processing and the application of laminarin to a certain extent.
Summary of the invention
The invention provides a kind of technical maturity, the applicable method of extracting laminarin from sea-tangle of producing.
The present invention is achieved through the following technical solutions:
A method of extracting laminarin from sea-tangle, comprises the steps:
A. raw material is processed: after sea-tangle is drying, pulverize;
B. ultrasonication: in the Kelp Powder after pulverizing, add 10-20 pure water doubly, ultrasonication 20-40min, operative temperature 70-85 ℃, power is 200-350W, filtrate is collected in filtration after completing;
C. enzymolysis: add the trypsinase of filtrate weight 1.5-2.5%, stir, 35-45 ℃ of enzymolysis 1-3h, adjust pH is to 3.0-5.0, the stomach en-that adds again filtrate weight 1.5-2.5%, stirs, 35-45 ℃ of enzymolysis 1-3h, then boiling water bath 10-20min, filters, and collects filtrate;
D. decolouring: add in the filtrate obtaining to step c and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, and add the activated carbon of filtrate weight 0.5% in filtrate, 80 ℃ of insulation 25-35min decolour, and suction filtration, collects filtrate;
E. except albumen: add in the filtrate obtaining to steps d and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, in filtrate, slowly add Potassium ethanoate dilute solution to final concentration 2mol/L, 4 ℃ of standing over night, centrifugal 10-15min, precipitation is carried out lyophilize and is obtained finished product.
Preferably, in step a, be crushed to 40-60 order.
Preferably, in step b, in the Kelp Powder after pulverizing, add 15 times of pure water.
Preferably, ultrasonic treatment time 30min in step b.
Preferably, 80 ℃ of ul-trasonic irradiation temperature in step b.
Preferably, ultrasonic power 300W in step b.
Preferably, in step c, trypsinase or pepsic hydrolysis temperature are 40 ℃.
Preferably, in step c, trypsinase or pepsic enzymolysis time are 2h.
Preferably, in step c, trypsinase or pepsic consumption are 2.0%.
Preferably, in steps d, add be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95% to step c in the filtrate obtaining, 4 ℃ of standing 4h, filter, and precipitation adds in step c, again enzymolysis.
Preferably, in step e, centrifugal is 5000r/min.
Adopt the laminarin that method obtains that obtains of the present invention, there is following proterties:
The slightly yellowy powder of white, easily moisture absorption, soluble in water.
Adopt method of the present invention to take sea-tangle as raw material extraction laminarin, compared with prior art, advantage of the present invention:
1, the bright method of this law is prepared laminarin by modern technique Integrated usings such as ultrasonic wave, proteolysis, alcohol precipitation, lyophilizes, technical maturity, reliable, and this method can be cost-saving, reduce energy consumption, improves extraction efficiency.
2, the present invention is on raw material is processed, and takes after ultrasonication enzymolysis again, can keep well the proterties of polysaccharide protein, and prior art adopts long-time high-temperature heating treatment, causes the sex change of raw material gel, polysaccharide protein.
3, use two enzymes to decompose laminarin, first adopt after ultrasonication, while adding prozyme again, just can carry out thorough enzymolysis to laminarin albumen, without micronizing and the nano-pulverization of carrying out raw material, simultaneously, in the present invention, the filter residue after enzymolysis decolouring carries out enzymolysis again, thus the color and luster of cost-saving and effective assurance product.
4, adopt Potassium ethanoate to remove albumen, safe and easy recycling, remove the high easily-controlled reaction conditions of albumen rate, avoided traditional Sevage method to remove the use of high toxicity organic solvents, chloroform in albumen, also avoided adopting hydrogen peroxide except egg from and the target product Structural Isomerism that causes causes laminarin content to decline, produce relatively safely, product content is higher.
5, adopt lyophilize be dried laminarin, the product crystalline form that obtains is better, the content of product can up to more than 98.5%, color and luster is good.
Embodiment
With embodiment, the invention will be further described below, but the present invention is not limited to these embodiment.
Embodiment 1:
A method of extracting laminarin from sea-tangle, comprises the steps:
A. raw material is processed: get Fresh Laminaria Japonica 10kg, after drying, be crushed to 40 orders;
B. ultrasonication: in the Kelp Powder after pulverizing, add the pure water of 10 times, ultrasonication 20min, 80 ℃ of operative temperatures, power is 300W, filtrate is collected in filtration after completing;
C. enzymolysis: add the trypsinase of slurry weight 1.5%, stir, 45 ℃ of enzymolysis 1h, adjust pH to 3.0, then add the stomach en-of slurry weight 2.5%, and stir, 35 ℃ of enzymolysis 3h, then boiling water bath 10min, filters, and collects filtrate;
D. decolouring: add in the filtrate obtaining to step c and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, and add the activated carbon of filtrate weight 0.5% in filtrate, 80 ℃ of insulation 25min decolour, and suction filtration, collects filtrate;
E. except albumen: add in the filtrate obtaining to steps d and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, in filtrate, slowly add Potassium ethanoate dilute solution to final concentration 2mol/L, 4 ℃ of standing over night, centrifugal, 5000r/min, 10min, precipitation is carried out lyophilize and is obtained laminarin 2.18kg, laminarin content 98.65%.
Embodiment 2:
A method of extracting laminarin from sea-tangle, comprises the steps:
A. raw material is processed: get the sea-tangle 10kg after drying, be crushed to 60 orders;
B. ultrasonication: in the Kelp Powder after pulverizing, add the pure water of 20 times, ultrasonication 40min, 70 ℃ of operative temperatures, power is 350W, filtrate is collected in filtration after completing;
C. enzymolysis: add the trypsinase of slurry weight 2.5%, stir, 35 ℃ of enzymolysis 3h, adjust pH to 5.0, then add the stomach en-of slurry weight 1.5%, and stir, 45 ℃ of enzymolysis 1h, then boiling water bath 20min, filters, and collects filtrate;
D. decolouring: add in the filtrate obtaining to step c and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, and add the activated carbon of filtrate weight 0.5% in filtrate, 80 ℃ of insulation 35min insulation of decolour, suction filtration, collection filtrate;
E. except albumen: add in the filtrate obtaining to steps d and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, in filtrate, slowly add Potassium ethanoate dilute solution to final concentration 2mol/L, 4 ℃ of standing over night, centrifugal, 5000r/min, 15min, precipitation is carried out lyophilize and is obtained laminarin 2.02kg, laminarin content 98.52%.
Embodiment 3:
A method of extracting laminarin from sea-tangle, comprises the steps:
A. raw material is processed: get Fresh Laminaria Japonica 10kg, after drying, be crushed to 60 orders;
B. ultrasonication: in the Kelp Powder after pulverizing, add the pure water of 15 times, ultrasonication 30min, 85 ℃ of operative temperatures, power is 200W, filtrate is collected in filtration after completing;
C. enzymolysis: add the trypsinase of slurry weight 2.0%, stir, 40 ℃ of enzymolysis 2h, adjust pH to 4.0, then add the stomach en-of slurry weight 2.0%, and stir, 40 ℃ of enzymolysis 2h, then boiling water bath 15min, filters, and collects filtrate;
D. decolouring: add in the filtrate obtaining to step c and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, the activated carbon that adds filtrate weight 0.5% in filtrate, 80 ℃ of insulation 30min insulation of decolouring, suction filtration, collect filtrate, filter residue returns to step c enzymolysis again;
E. except albumen: add in the filtrate obtaining to steps d and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, in filtrate, slowly add Potassium ethanoate dilute solution to final concentration 2mol/L, 4 ℃ of standing over night, centrifugal, 5000r/min, 12min, precipitation is carried out lyophilize and is obtained laminarin 2.22kg, laminarin content 99.11%.
Claims (10)
1. from sea-tangle, extract a method for laminarin, comprise the steps:
A. raw material is processed: after sea-tangle is drying, pulverize;
B. ultrasonication: in the Kelp Powder after pulverizing, add 10-20 pure water doubly, ultrasonication 20-40min, operative temperature 70-85 ℃, power is 200-350W, filtrate is collected in filtration after completing;
C. enzymolysis: add the trypsinase of filtrate weight 1.5-2.5%, stir, 35-45 ℃ of enzymolysis 1-3h, adjust pH is to 3.0-5.0, the stomach en-that adds again filtrate weight 1.5-2.5%, stirs, 35-45 ℃ of enzymolysis 1-3h, then boiling water bath 10-20min, filters, and collects filtrate;
D. decolouring: add in the filtrate obtaining to step c and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, and add the activated carbon of filtrate weight 0.5% in filtrate, 80 ℃ of insulation 25-35min decolour, and suction filtration, collects filtrate;
E. except albumen: add in the filtrate obtaining to steps d and be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95%, 4 ℃ of standing 4h, filter, in filtrate, slowly add Potassium ethanoate dilute solution to final concentration 2mol/L, 4 ℃ of standing over night, centrifugal 10-15min, precipitation is carried out lyophilize and is obtained finished product.
2. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in step a, be crushed to 40-60 order.
3. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in the Kelp Powder after pulverizing in step b, add 15 times of pure water.
4. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: ultrasonic treatment time 30min in step b.
5. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in step b, ul-trasonic irradiation temperature is 80 ℃.
6. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: ultrasonic power 300W in step b.
7. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in step c, trypsinase or pepsic hydrolysis temperature are 40 ℃.
8. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in step c, trypsinase or pepsic enzymolysis time are 2h.
9. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in step c, trypsinase or pepsic consumption are 2.0%.
10. the method for extracting laminarin from sea-tangle according to claim 1, is characterized in that: in steps d, add be equivalent to the ethanol that the mass concentration of 2 times of filtrate weight is 95% to step c in the filtrate obtaining, 4 ℃ of standing 4h, filter, precipitation adds in step c, again enzymolysis.
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104387497A (en) * | 2014-12-19 | 2015-03-04 | 桂林市和胤祥新型材料有限公司 | Extraction method of laminarin with antineoplastic activity |
| CN104446820A (en) * | 2014-12-19 | 2015-03-25 | 桂林市和胤祥新型材料有限公司 | Kelp-added ganoderma lucidum substitute cultivation culture medium |
| CN104479042A (en) * | 2014-12-19 | 2015-04-01 | 桂林市和胤祥新型材料有限公司 | Method for extracting laminarin from kelp |
| CN104479041A (en) * | 2014-12-19 | 2015-04-01 | 桂林市和胤祥新型材料有限公司 | Preparation method of laminarin with anti-tumor activity |
| CN104478610A (en) * | 2014-12-19 | 2015-04-01 | 桂林市和胤祥新型材料有限公司 | Kelp-containing enoki mushroom substitute cultivation medium |
| CN104710541A (en) * | 2015-03-31 | 2015-06-17 | 广西还珠海洋生物科技有限公司 | Method for preparing laminarin from kelp |
| CN105920050A (en) * | 2016-06-16 | 2016-09-07 | 广东海洋大学 | Preparation method and application of kelp extract for effectively reducing uric acid |
| CN106046188A (en) * | 2016-07-04 | 2016-10-26 | 青岛吉海营养科技有限公司 | Method for preparing fucoidin |
| CN106541482A (en) * | 2016-09-29 | 2017-03-29 | 阜南县星光工艺品有限公司 | A kind of preparation method of formaldehydeless fire-resistant-chipboard |
| CN107286267A (en) * | 2017-07-27 | 2017-10-24 | 福建农林大学 | One Polysaccharides From Laminaria Japonica Zn complex and its preparation and application |
| CN108420834A (en) * | 2018-06-07 | 2018-08-21 | 艾苛密（上海）健康科技股份有限公司 | Repair of cartilage marine algae extract and preparation method thereof |
| CN109776691A (en) * | 2018-12-28 | 2019-05-21 | 福建大昌生物科技实业有限公司 | A kind of novel processing technique for kelp |
| CN110240660A (en) * | 2019-06-21 | 2019-09-17 | 福建福瑞康信息技术有限公司 | The extraction process of one Polysaccharides From Laminaria Japonica |
| CN110922501A (en) * | 2019-10-24 | 2020-03-27 | 浙江工业大学 | Process for extracting sodium alginate by using ultrasonic-assisted complex enzyme |
| CN114539435A (en) * | 2022-01-17 | 2022-05-27 | 江南大学 | Three-method synergistic extraction process of laminarin |
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Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104479041B (en) * | 2014-12-19 | 2017-01-04 | 桂林市和胤祥新型材料有限公司 | A kind of preparation method of the laminarin with anti-tumor activity |
| CN104479042A (en) * | 2014-12-19 | 2015-04-01 | 桂林市和胤祥新型材料有限公司 | Method for extracting laminarin from kelp |
| CN104387497A (en) * | 2014-12-19 | 2015-03-04 | 桂林市和胤祥新型材料有限公司 | Extraction method of laminarin with antineoplastic activity |
| CN104479041A (en) * | 2014-12-19 | 2015-04-01 | 桂林市和胤祥新型材料有限公司 | Preparation method of laminarin with anti-tumor activity |
| CN104478610A (en) * | 2014-12-19 | 2015-04-01 | 桂林市和胤祥新型材料有限公司 | Kelp-containing enoki mushroom substitute cultivation medium |
| CN104446820A (en) * | 2014-12-19 | 2015-03-25 | 桂林市和胤祥新型材料有限公司 | Kelp-added ganoderma lucidum substitute cultivation culture medium |
| CN104710541A (en) * | 2015-03-31 | 2015-06-17 | 广西还珠海洋生物科技有限公司 | Method for preparing laminarin from kelp |
| CN105920050A (en) * | 2016-06-16 | 2016-09-07 | 广东海洋大学 | Preparation method and application of kelp extract for effectively reducing uric acid |
| CN106046188A (en) * | 2016-07-04 | 2016-10-26 | 青岛吉海营养科技有限公司 | Method for preparing fucoidin |
| CN106046188B (en) * | 2016-07-04 | 2018-12-21 | 青岛吉海营养科技有限公司 | A kind of preparation method of fucoidin |
| CN106541482A (en) * | 2016-09-29 | 2017-03-29 | 阜南县星光工艺品有限公司 | A kind of preparation method of formaldehydeless fire-resistant-chipboard |
| CN107286267A (en) * | 2017-07-27 | 2017-10-24 | 福建农林大学 | One Polysaccharides From Laminaria Japonica Zn complex and its preparation and application |
| CN108420834A (en) * | 2018-06-07 | 2018-08-21 | 艾苛密（上海）健康科技股份有限公司 | Repair of cartilage marine algae extract and preparation method thereof |
| CN108420834B (en) * | 2018-06-07 | 2021-04-23 | 艾苛密（上海）健康科技股份有限公司 | Seaweed extract for cartilage repair and preparation method thereof |
| CN109776691A (en) * | 2018-12-28 | 2019-05-21 | 福建大昌生物科技实业有限公司 | A kind of novel processing technique for kelp |
| CN110240660A (en) * | 2019-06-21 | 2019-09-17 | 福建福瑞康信息技术有限公司 | The extraction process of one Polysaccharides From Laminaria Japonica |
| CN110922501A (en) * | 2019-10-24 | 2020-03-27 | 浙江工业大学 | Process for extracting sodium alginate by using ultrasonic-assisted complex enzyme |
| CN114539435A (en) * | 2022-01-17 | 2022-05-27 | 江南大学 | Three-method synergistic extraction process of laminarin |
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Application publication date: 20140723 |