CN106832027A - The extracting method of gumbo polysaccharide - Google Patents
The extracting method of gumbo polysaccharide Download PDFInfo
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- CN106832027A CN106832027A CN201611251131.2A CN201611251131A CN106832027A CN 106832027 A CN106832027 A CN 106832027A CN 201611251131 A CN201611251131 A CN 201611251131A CN 106832027 A CN106832027 A CN 106832027A
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- C—CHEMISTRY; METALLURGY
- 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 present invention provides a kind of extracting method of gumbo polysaccharide, concretely comprises the following steps:1)Pretreatment;2)Degreasing;3)Enzymolysis;4)Purifying;5)De- albumen.Have the beneficial effect that and use complex enzyme zymohydrolysis, the mode of solubilization of microwave extracts polysaccharide.Enzymatic hydrolysis condition is gentle, will not destroy the structure of gumbo polysaccharide, it is kept activity higher;Various enzyme-linked synthase solutions, and have various zymoexciter activation complex enzymes, enzymolysis speed is fast and enzymolysis is more thorough, improves the recovery rate of gumbo polysaccharide;It is combined using solvent extraction and Microwave Extraction, improves polysaccharide extract rate;Chloroform and n-butanol can make protein denaturation and water insoluble, add chloroform and n-butanol to form jello in the polysaccharide solution containing protein, and isolating protein is removed by way of centrifugation;The purity of the gumbo polysaccharide for finally obtaining is very high, and recovery rate is high and can keep greater activity;Operating procedure is simple, with low cost, high financial profit.
Description
Technical field
The present invention relates to technical field of polysaccharide extraction, specifically a kind of extracting method of gumbo polysaccharide.
Background technology
Gumbo (Abelmoschus esculentus L .Moench) also known as okra, swordweed, Abelmoschus esculentus, hair
Eggplant, Sunset Abelmoschus Root, also referred to as " foreign capsicum " among the people.Africa is originated in, 20 beginnings of the century introduced China, and were more common in southern china by India.
Its edible portion is Fruit pod, is divided to two kinds of green and red, and the tender and crisp succulence of mouthfeel is lubricious oiliness, and fragrance is unique, and seed can be squeezed
Oil.Gumbo nature and flavor are light, cold, with relieving sore-throat, it is treating stranguria, it is lower breast, menstruation regulating the effect of.Gumbo contains abundant vitamin and mineral matter,
In the okra fruit of every 100 grams of gumbos, the about carrot of the vitamin C containing 4 milligrams, 1 .03 milligrams of vitamin E and 310 micrograms
Element.The zinc and selenium and other trace elements being rich in gumbo, it is helpful to enhancing human body cancer-resisting ability.Secondly, gumbo okra fruit
In have and stick glutinous liquid substance, this mucus contains the polysaccharide such as pectin and glutinous polysaccharide.Glutinous polysaccharide has enhancing Abwehrkraft des Koepers,
The glossy effect of membrana articulata and serous coat in human synovial chamber is safeguarded, accumulation of the lipid material on ductus arteriosus wall is cut down, it is to avoid liver
In dirty and kidney the effects such as adesmosis.Furthermore, gumbo water content is high, and seldom, every 100 grams of gumbo okra fruits are comprised only fat
0 .1 grams of fat, is well suited for wanting the Ms of weight-reducing, and its vitamin C and dietary fiber for being rich in, moreover it is possible to make skin
It is tender white.Additionally, the stickum of okra, can promote gastrointestinal peristalsis, it is beneficial to aid digestion, beneficial stomach;The stickum of gumbo
In containing 50% soluble cellulose, favourable defaecation, toxin expelling, anti-cancer can effectively reduce serum cholesterols, prevention of cardiovascular
Disease, hypoxia-bearing capability can be improved;The pectin and polysaccharide contained in okra can have protect liver effect, anti-to treat constipation;Meanwhile, okra
Not only calcium content is suitable with fresh milk, and calcium absorptivity in 50-60%, be preferable calcium source higher than 1 times of milk;Okra is
Low energy food, and flavones content is 2 .8%, delaying sanility, with anti-oxidant, anti-aging effect prevents and treats diabetes.
Extraction now for polysaccharide is much studied, and prior art such as Authorization Notice No. are in CN104311691B
State's patent of invention, discloses a kind of extracting method of bitter melon polysaccharide, including following steps:Balsam pear is cleaned up and dried,
Balsam pear powder is ground into again, papain and neutral proteinase are added in balsam pear powder, digested under microwave condition, it is dense after enzymolysis
Contract decolouring, centrifugation takes supernatant concentration alcohol precipitation, then is centrifuged, cleaned using ethanol and precipitated, after freeze-drying and crush, obtain bitter
Melon polysaccharide.Polysaccharide is extracted from balsam pear using the method, using enzyme effect mild condition, using a variety of enzymes by balsam pear
Fat, cellulase hydrolysis, improve the utilization rate of balsam pear by Polyose extraction therein out.The method does not take off albumen, polysaccharide
Purity is not high, and recovery rate also has much room for improvement.Prior art such as Authorization Notice No. is the Chinese invention patent of CN104844720B,
A kind of separating and extracting process of hundred tails gracilis polysaccharide is disclosed, is comprised the following steps:Crushed after fresh hundred tail is joined into cleaning, drying;
With petroleum ether backflow degreasing;Addition distilled water is extracted, and is extracted and is terminated to merge Aqueous extracts;Aqueous extracts Buchner funnel suction filtration, filtrate
Stood after alcohol precipitation, be centrifuged, removing supernatant, precipitation uses distilled water constant volume;Decolourize to purify, concentrate, obtain Thick many candies liquid;Thick many candies liquid
In mass ratio 3:1 ratio adds the mixed liquor of chloroform and n-butanol, and supernatant liquor is taken after shaking, add ethanol, standing, from
The heart, obtains the tail gracilis polysaccharide of de- albumen hundred;Washing, freeze-drying is obtained final product.The method can obtain that purity is high, hundred tail of quality better ginseng is more
Sugar, method is simple, easy to operate, great market popularization value.The method extracts polysaccharide only with water extraction, does not take other modes
Assisted extraction, recovery rate is not high.
The content of the invention
High it is an object of the invention to provide a kind of recovery rate, the active polysaccharide of extraction is strong, purity gumbo polysaccharide high
Extracting method.
The present invention is directed to the problem mentioned in background technology, and the technical scheme taken is:
The extracting method of gumbo polysaccharide, concrete operation step:
1)Pretreatment:Fresh gumbo is dried in air blower, temperature is 50 ~ 70 DEG C, and drying time is 0.8 ~ 1.5h, until the autumn
Certain herbaceous plants with big flowers reclaimed water is divided into 3 ~ 8%, and 40 ~ 100 mesh sieves are crossed in grinding;
2)Degreasing:It is 1 that solid-liquid ratio is pressed in gumbo powder:40~1:80 add petroleum ether, in apparatus,Soxhlet's, at 40 ~ 60 DEG C
Backflow 2 ~ 4h of degreasing, filtration drying obtains degreasing gumbo powder;
3)Enzymolysis:Solid-liquid ratio 1 is pressed in degreasing gumbo powder:30~1:60 add distilled water, stirring, and stir speed (S.S.) is 200 ~ 500r/
min.Enzymolysis agent is added, enzymolysis agent composition and its weight portion are 2 ~ 6 parts of cellulases, 1 ~ 5 part of pectase, 0.4 ~ 0.7 part of ox pancreas
Protease, 0.2 ~ 0.5 part of calcium chloride, 0.01 ~ 0.03 part of APES, 0.02 ~ 0.04 part of cysteine, 0.01 ~
0.04 part of Leaf Acetate and 0.03 ~ 0.05 part of 2- pyrrolidine acetic acid.Hydrolysis temperature is 40 ~ 55 DEG C, and pH is 5.2 ~ 6.8, enzymolysis
Time is 0.1 ~ 0.4h.Solubilization of microwave, microwave power is 200 ~ 400W, and frequency is 500 ~ 1MHz, and action time is 30 ~ 50min.
Enzymolysis is gone out enzyme after terminating with boiling water bath, and the enzyme time of going out is for 10 ~ 20min.Filtering, filtrate through be concentrated in vacuo to original volume 1/4 ~
1/3 obtains gumbo Thick many candies concentrate.Enzymatic hydrolysis condition is gentle, will not destroy the structure of gumbo polysaccharide, it is kept work higher
Property;Various enzyme-linked synthase solutions, and have various zymoexciter activation complex enzymes, enzymolysis speed is fast and enzymolysis is more thorough, improves gumbo
The recovery rate of polysaccharide;It is combined using solvent extraction and Microwave Extraction, is carried out as heater in itself using heating object
Heating, not by conduction of heat, can be such that internal batch temperature improves rapidly, and the heat time shortens, while being made using distilled water
Be Extraction solvent, with it is cheap, nontoxic, be easily recycled the characteristics of, greatly reduce production cost;
4)Purifying:In mass ratio 1 in gumbo Thick many candies concentrate:3~1:5 add 95% ethanol, stand, and supernatant is removed in centrifugation
Liquid, in mass ratio 1 in precipitation:10~1:30 add distilled water, are purified with AB-8 resin decolorizations;
5)De- albumen:Add mass ratio 4:1~6:1 chloroform and the mixed liquor of n-butanol, addition are more for gumbo after purification of decolourizing
The 1/2 ~ 1/4 of sugared concentrate quality.Filter to take supernatant after stirring, add 95% ethanol, stand centrifugation, collection is precipitated and uses 95%
Ethanol is cleaned repeatedly, freeze-drying, and is crushed at 1 ~ 5 DEG C, obtains gumbo polysaccharide powder.Chloroform and n-butanol can make protein
Denaturation and it is water insoluble, add chloroform and n-butanol to form jello in the polysaccharide solution containing protein, by what is be centrifuged
Mode removes isolating protein.The purity of the gumbo polysaccharide for finally obtaining is very high, and recovery rate is high and can keep greater activity.
Compared with prior art, the advantage of the invention is that:Enzymatic hydrolysis condition is gentle, will not destroy the structure of gumbo polysaccharide,
It is set to keep activity higher;Various enzyme-linked synthase solutions, and have various zymoexciter activation complex enzymes, enzymolysis speed is fast and digests
More thoroughly, the recovery rate of gumbo polysaccharide is improved;It is combined using solvent extraction and Microwave Extraction, is made in itself using heating object
Heated for heater, not by conduction of heat, internal batch temperature can be made to improve rapidly, the heat time is shortened, together
Shi Caiyong distilled water as Extraction solvent, with it is cheap, nontoxic, be easily recycled the characteristics of, greatly reduce and be produced into
This;Chloroform and n-butanol can make protein denaturation and water insoluble, and chloroform and positive fourth are added in the polysaccharide solution containing protein
Alcohol can form jello, and isolating protein is removed by way of centrifugation;The purity of the gumbo polysaccharide for finally obtaining is very high, and recovery rate is high
And greater activity can be kept;Operating procedure is simple, with low cost, high financial profit.
Specific embodiment
The present invention program is described further below by embodiment:
Embodiment 1:
The extracting method of gumbo polysaccharide, concrete operation step:
1)Pretreatment:Fresh gumbo is dried in air blower, temperature is 50 ~ 70 DEG C, and drying time is 0.8 ~ 1.5h, until the autumn
Certain herbaceous plants with big flowers reclaimed water is divided into 3 ~ 8%, and 40 ~ 100 mesh sieves are crossed in grinding;
2)Degreasing:It is 1 that solid-liquid ratio is pressed in gumbo powder:40~1:80 add petroleum ether, in apparatus,Soxhlet's, at 40 ~ 60 DEG C
Backflow 2 ~ 4h of degreasing, filtration drying obtains degreasing gumbo powder;
3)Enzymolysis:Solid-liquid ratio 1 is pressed in degreasing gumbo powder:30~1:60 add distilled water, stirring, and stir speed (S.S.) is 200 ~ 500r/
min.Enzymolysis agent is added, enzymolysis agent composition and its weight portion are 2 ~ 6 parts of cellulases, 1 ~ 5 part of pectase, 0.4 ~ 0.7 part of ox pancreas
Protease, 0.2 ~ 0.5 part of calcium chloride, 0.01 ~ 0.03 part of APES, 0.02 ~ 0.04 part of cysteine, 0.01 ~
0.04 part of Leaf Acetate and 0.03 ~ 0.05 part of 2- pyrrolidine acetic acid.Hydrolysis temperature is 40 ~ 55 DEG C, and pH is 5.2 ~ 6.8, enzymolysis
Time is 0.1 ~ 0.4h.Solubilization of microwave, microwave power is 200 ~ 400W, and frequency is 500 ~ 1MHz, and action time is 30 ~ 50min.
Enzymolysis is gone out enzyme after terminating with boiling water bath, and the enzyme time of going out is for 10 ~ 20min.Filtering, filtrate through be concentrated in vacuo to original volume 1/4 ~
1/3 obtains gumbo Thick many candies concentrate.Enzymatic hydrolysis condition is gentle, will not destroy the structure of gumbo polysaccharide, it is kept work higher
Property;Various enzyme-linked synthase solutions, and have various zymoexciter activation complex enzymes, enzymolysis speed is fast and enzymolysis is more thorough, improves gumbo
The recovery rate of polysaccharide;It is combined using solvent extraction and Microwave Extraction, is carried out as heater in itself using heating object
Heating, not by conduction of heat, can be such that internal batch temperature improves rapidly, and the heat time shortens, while being made using distilled water
Be Extraction solvent, with it is cheap, nontoxic, be easily recycled the characteristics of, greatly reduce production cost;
4)Purifying:In mass ratio 1 in gumbo Thick many candies concentrate:3~1:5 add 95% ethanol, stand, and supernatant is removed in centrifugation
Liquid, in mass ratio 1 in precipitation:10~1:30 add distilled water, are purified with AB-8 resin decolorizations;
5)De- albumen:Add mass ratio 4:1~6:1 chloroform and the mixed liquor of n-butanol, addition are more for gumbo after purification of decolourizing
The 1/2 ~ 1/4 of sugared concentrate quality.Filter to take supernatant after stirring, add 95% ethanol, stand centrifugation, collection is precipitated and uses 95%
Ethanol is cleaned repeatedly, freeze-drying, and is crushed at 1 ~ 5 DEG C, obtains gumbo polysaccharide powder.Chloroform and n-butanol can make protein
Denaturation and it is water insoluble, add chloroform and n-butanol to form jello in the polysaccharide solution containing protein, by what is be centrifuged
Mode removes isolating protein.The purity of the gumbo polysaccharide for finally obtaining is very high, and recovery rate is high and can keep greater activity.
Embodiment 2:
The extracting method of gumbo polysaccharide, most preferably operating procedure:
1)Pretreatment:Fresh gumbo is dried in air blower, temperature is 60 DEG C, and drying time is 1h, until moisture in gumbo
It is 5%, 50 mesh sieves are crossed in grinding;
2)Degreasing:It is 1 that solid-liquid ratio is pressed in gumbo powder:60 add petroleum ether, in apparatus,Soxhlet's, in the degreasing that flowed back at 50 DEG C
3h, filtration drying obtains degreasing gumbo powder;
3)Enzymolysis:Solid-liquid ratio 1 is pressed in degreasing gumbo powder:40 add distilled water, stir, and stir speed (S.S.) is 300r/min.Add
Enzymolysis agent, most preferably enzymolysis agent composition and its weight portion are 5 parts of cellulases, 3 parts of pectases, 0.5 part of bovine trypsin, 0.4
Part calcium chloride, 0.02 part of APES, 0.03 part of cysteine, 0.02 part of Leaf Acetate and 0.05 part of 2- pyrroles
Alkane acetic acid.Hydrolysis temperature is 45 DEG C, and pH is 6.2, and enzymolysis time is 0.2h.Solubilization of microwave, microwave power is 300W, and frequency is
800MHz, action time is 30min.Enzymolysis is gone out enzyme after terminating with boiling water bath, and the enzyme time of going out is for 15min.Filtering, filtrate is through vacuum
It is concentrated into the 1/4 of original volume and obtains gumbo Thick many candies concentrate.Enzymatic hydrolysis condition is gentle, will not destroy the structure of gumbo polysaccharide,
It is set to keep activity higher;Various enzyme-linked synthase solutions, and there have the composition such as Leaf Acetate and 2- pyrrolidine acetic acids to activate to be compound
Enzyme, synergy enzymolysis, enzymolysis speed is fast and enzymolysis is more thorough, improves the recovery rate of gumbo polysaccharide;Carried using solvent extraction and microwave
Take and be combined, heated as heater in itself using heating object, not by conduction of heat, can make inside material
Temperature is improved rapidly, and the heat time shortens, while using distilled water as Extraction solvent, with it is cheap, nontoxic, be easily recycled again
The characteristics of utilization, greatly reduce production cost;
4)Purifying:In mass ratio 1 in gumbo Thick many candies concentrate:4 add 95% ethanol, stand, and supernatant is removed in centrifugation,
In mass ratio 1 in precipitation:18 add distilled water, are purified with AB-8 resin decolorizations;
5)De- albumen:Add mass ratio 5:1 chloroform and the mixed liquor of n-butanol, addition are dense for the gumbo polysaccharide after purification that decolourizes
The 1/3 of contracting liquid quality.Filter to take supernatant after stirring, add 95% ethanol, stand centrifugation, collection precipitate and with 95% ethanol repeatedly
Cleaning, freeze-drying, and crushed at 1 ~ 5 DEG C, obtain gumbo polysaccharide powder.Chloroform and n-butanol can make protein denaturation without
Water is dissolved in, adds chloroform and n-butanol to form jello in the polysaccharide solution containing protein, removed by way of centrifugation
Protein.The purity of the gumbo polysaccharide for finally obtaining is very high, and recovery rate is high and can keep greater activity.
Routine operation in operating procedure of the invention is well known to those skilled in the art, and is not repeated herein.
Embodiment described above has been described in detail to technical scheme, it should be understood that the above is only
It is specific embodiment of the invention, is not intended to limit the invention, all any modifications made in spirit of the invention,
Supplement or similar fashion replacement etc., should be included within the scope of the present invention.
Claims (9)
1. the extracting method of gumbo polysaccharide, it is characterised in that following steps:
1)Pretreatment:Fresh gumbo is dried, grinding, sieving;
2)Degreasing:Organic solution is added in gumbo powder, flow back degreasing, obtains degreasing gumbo powder;
3)Enzymolysis:Distilled water is added in degreasing gumbo powder, is stirred, add enzymolysis agent, solubilization of microwave digests the enzyme that gone out after terminating,
4)4)Filtering, filtrate is concentrated to give gumbo Thick many candies concentrate;
5)Purifying:Ethanol is added in gumbo Thick many candies concentrate, is stood, supernatant is removed in centrifugation, and distillation is added in precipitation
Water, is purified with resin decolorization;
6)De- albumen:De- protein liquid is added, supernatant is filtered to take after stirring, add ethanol, stand centrifugation, collection is precipitated and uses second
Alcohol is cleaned repeatedly, freeze-drying, and is crushed at low temperature, obtains gumbo polysaccharide powder.
2. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Organic solution in described step 2
It is petroleum ether, solid-liquid ratio is 1:40~1:80, in apparatus,Soxhlet's, in the 2 ~ 4h of degreasing that flowed back at 40 ~ 60 DEG C.
3. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Distilled water is pressed in described step 3
Solid-liquid ratio 1:30~1:60 add, and stir speed (S.S.) is 200 ~ 500r/min.
4. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:In described step 3 enzymolysis agent into
Part and its weight portion be 2 ~ 6 parts of cellulases, 1 ~ 5 part of pectase, 0.4 ~ 0.7 part of bovine trypsin, 0.2 ~ 0.5 part of calcium chloride,
0.01 ~ 0.03 part of APES, 0.02 ~ 0.04 part of cysteine, 0.01 ~ 0.04 part of Leaf Acetate and 0.03 ~
0.05 part of 2- pyrrolidine acetic acid.
5. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Hydrolysis temperature in described step 3
It it is 40 ~ 55 DEG C, pH is 5.2 ~ 6.8, enzymolysis time is 0.1 ~ 0.4h.
6. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Microwave power in described step 3
It is 200 ~ 400W, frequency is 500 ~ 1MHz, and action time is 30 ~ 50min.
7. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Boiling water bath is used in described step 3
Go out enzyme, and the enzyme time of going out is for 10 ~ 20min.
8. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Protein liquid is taken off in described step 3
It is mass ratio 4:1~6:1 chloroform and the mixed liquor of n-butanol, addition are gumbo polysaccharide concentrate quality after purification of decolourizing
1/2~1/4。
9. the extracting method of gumbo polysaccharide according to claim 1, it is characterised in that:Ethanol in described step 4 and 5
It is 95% ethanol.
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Cited By (6)
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CN106890203A (en) * | 2017-04-20 | 2017-06-27 | 广州市天河华南发展有限公司 | A kind of gumbo extract and its preparation method and application |
CN109019742A (en) * | 2018-07-11 | 2018-12-18 | 李玉茹 | A kind of sewage-treating agent and preparation method thereof of efficient absorption heavy metal |
CN109156585A (en) * | 2018-08-06 | 2019-01-08 | 四川大学 | A kind of gumbo gel candy and preparation method thereof |
CN109234335A (en) * | 2018-11-14 | 2019-01-18 | 浙江海洋大学 | A kind of preparation method of polysaccharide in tabasheer rich in galactofuranose |
CN109321554A (en) * | 2018-11-30 | 2019-02-12 | 广西南宁天绿生物制品有限公司 | A kind of production method of the dedicated papain of beer |
CN116178585A (en) * | 2023-03-20 | 2023-05-30 | 江苏沿江地区农业科学研究所 | Combined method for extracting okra polysaccharide |
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CN106890203A (en) * | 2017-04-20 | 2017-06-27 | 广州市天河华南发展有限公司 | A kind of gumbo extract and its preparation method and application |
CN109019742A (en) * | 2018-07-11 | 2018-12-18 | 李玉茹 | A kind of sewage-treating agent and preparation method thereof of efficient absorption heavy metal |
CN109156585A (en) * | 2018-08-06 | 2019-01-08 | 四川大学 | A kind of gumbo gel candy and preparation method thereof |
CN109234335A (en) * | 2018-11-14 | 2019-01-18 | 浙江海洋大学 | A kind of preparation method of polysaccharide in tabasheer rich in galactofuranose |
CN109234335B (en) * | 2018-11-14 | 2021-08-20 | 浙江海洋大学 | Preparation method of polysaccharide rich in galactofuranose in tabasheer |
CN109321554A (en) * | 2018-11-30 | 2019-02-12 | 广西南宁天绿生物制品有限公司 | A kind of production method of the dedicated papain of beer |
CN109321554B (en) * | 2018-11-30 | 2021-11-09 | 广西南宁天绿生物制品有限公司 | Production method of special papain for beer |
CN116178585A (en) * | 2023-03-20 | 2023-05-30 | 江苏沿江地区农业科学研究所 | Combined method for extracting okra polysaccharide |
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