CN101069562A - Zymolysis process for preparing high-immunity regulation activity wolfberry polysaccharides using wolfberry wine residues - Google Patents
Zymolysis process for preparing high-immunity regulation activity wolfberry polysaccharides using wolfberry wine residues Download PDFInfo
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Abstract
The present invention provides a process method for producing high immunoregulatory activity lyceum polysaccharide by utilizing enzymolysis method and by using lyceum berry residue obtained after the lyceum berry is soacked in the white liquor as as raw material. Said process method includes the following steps: adopting water extraction alcohol precipitation method to extract primary polysaccharide from lyceum berry liquor residue, then adding amylase liquor whose concentration is 0.004-0.01%, making hydrolysis for 15-40 min at 30-50deg.C, boiling hydrolysate for 10 min, then adding 95% ethyl alcohol to make precipitation (alcohol precipitation concentration is 80%), removing low-molecular weight soluble saccharide, standing still for 10-18h, making centrifugal separation for 15 min by adopting 3000r/min, taking out precipitate and vacuum drying so as to obtain the invented brown high immunoregulatory activity enzymatic lycium polysaccharide.
Description
Technical field
The present invention relates to the enzymolysis process technology of polysaccharide, particularly a kind of technology for preparing high physiologically active enzymolysis LBP-X.It is to change the LBP-X molecular weight by zymolysis technique, thereby further improves the immunoregulatory activity of LBP-X.
Background technology
The fruit of Chinese wolfberry is traditional simply Chinese medicine, have tonify the liver and kidney, the effect of nourishing liver for improving eyesight.Along with pharmacology, nutrition progress of research to the matrimony vine main active, it has been recognized that the composition that plays a major role in the fruit of Chinese wolfberry is a polysaccharide, it is in nospecific immunity, and there is the regulating action of highly significant the specific immunity aspect, is suppressing tumour, delay senility, AFL, hypoglycemic, reducing blood lipid, antifatigue, aspect effects such as promotion growth are also very outstanding.Studies show that its physiologically active and molecular weight are in close relations.
The fruit of Chinese wolfberry generally is used for steeping medicinal liquor, because LBP-X is insoluble to ethanol, therefore, most LBP-Xs residue in the matrimony vine schlempe.And the first polysaccharide of the matrimony vine of adopting general water extraction and alcohol precipitation method to extract, immunoregulatory activity is not high.
The present invention is a raw material with the matrimony vine schlempe, utilizes zymolysis technique, and the matrimony vine that water extraction and alcohol precipitation method is made just polysaccharide carries out reasonable enzymolysis, and in conjunction with the Ergonomy contrast experiment, optimization stronger enzymolysis LBP-X and the preparation technology thereof of immunoregulatory activity.
Summary of the invention
The present invention proposes a kind of process of utilizing enzymatic isolation method to produce high-immunity regulation activity enzymolysis LBP-X.Its production technology is:
(1) with the oven dry of matrimony vine schlempe, wear into powder just, standby,
(2) add 6～9 times of amounts (solid-to-liquid ratio) water, in 70～90 ℃ of extractions 1～3 time, each 1～3h filters,
(3) merge whole filtrates, 55～75 ℃ to be evaporated to original volume 1/10 standby,
(4) in concentrate, add 95% ethanol, make pure content reach 80%, leave standstill and separated out precipitation in 10～18 hours, get pure hypostasis and get pale brown look to the first polysaccharide (LBP-X content 60～65%, weight average molecular weight 1～40,000) of dark-brown matrimony vine in 40～60 ℃ of vacuum drying,
(5) get matrimony vine polysaccharide just, be dissolved in water, be mixed with LBP-X concentration and be 2.4～3.6% first polysaccharide solution,
(6) get polysaccharide solution just, add the carbohydrase liquid (concentration is 0.004～0.01%) of 1/5 volume, in 30～50 ℃ of hydrolysis 15～40min, hydrolyzate boils 10min, with enzyme-deactivating,
(7) adding 95% ethanol concentration of alcohol to the solution to hydrolyzate is 80%, produces precipitation.Left standstill 10～18 hours, the centrifugal 15min of 3000r/min, taking precipitate,
(8) sediment promptly gets brown high-immunity regulation activity enzymolysis LBP-X (weight average molecular weight 1～30,000) in 40～60 ℃ of vacuum drying.
Carbohydrase liquid join method: (powder of 10000～50000 units/g) is with buffer solution (pH4.5) dissolving (enzyme liquid concentration 0.004～0.01%) to get the commodity purchased carbohydrase
Buffer solution join method: get sodium acetate 18g and add glacial acetic acid 9.8mL, thin up promptly gets (concentration C to 1000mL again
HAc=0.343mol/L)
With the enzymolysis LBP-X that the present invention makes, immunoregulatory activity height, it is as follows to record data by mouse carbon particle clearance and spleen index contrast experiment:
Table 1 enzymolysis LBP-X effect experiment table
* p＜0.05**p＜0.01 is all compared with model group
From finding out enzymolysis LBP-X carbon particle clearance index (0.027007 ± 0.006737), spleen index (0.004976 ± 0.001292) is greater than LBP-X (carbon particle clearance index 0.011939 ± 0.001954, the immunoregulatory activity height of enzymolysis LBP-X is described spleen index 0.003816 ± 0.000462).
The present invention is to have utilized the immunoregulatory activity of LBP-X and its molecular weight in close relations compared with prior art, by zymolysis technique, with matrimony vine just the polysaccharide enzymolysis with its molecular weight of suitable reduction, thereby further improve the immunoregulatory activity of LBP-X.It is that raw material prepares the high-immunity regulation activity LBP-X that technology of the present invention also is fit to the fruit of Chinese wolfberry.
The specific embodiment
Embodiment one: with 200 gram matrimony vine schlempe oven dry, wear into powder just, add 1.4L water, 80 ℃ are extracted 2 times, and each 3h merges whole filtrates.70 ℃ of concentrates that are evaporated to original volume 1/10.In concentrate, add 95% ethanol, make pure content reach 80%, leave standstill and separated out precipitation in 18 hours.The alcohol hypostasis is in 50 ℃ of vacuum drying, and gained matrimony vine just polysaccharide adds that 250mL is water-soluble to be separated, and adds 50mL concentration and be 0.004% carbohydrase liquid, in 40 ℃ of hydrolysis 30min.Hydrolyzate boils 10min, with enzyme-deactivating.Add 95% ethanol and make pure content reach 80%, leave standstill and separated out precipitation in 10～12 hours, centrifugal, 50 ℃ of vacuum drying promptly get brown high-immunity regulation activity enzymolysis LBP-X 40.4g.
Embodiment two: with 300 gram matrimony vine schlempe oven dry, wear into powder just, add 2.1L water, 75 ℃ are extracted 2 times, each 2h, merging filtrate.65 ℃ are evaporated to original volume 1/10.In concentrate, add 95% ethanol, make pure content reach 80%, leave standstill, separate out precipitation.Sediment is in 55 ℃ of vacuum drying, with the gained matrimony vine just polysaccharide add that 400mL is water-soluble to be separated, add 80mL concentration and be 0.007% carbohydrase liquid, in 50 ℃ of hydrolysis 20min.Hydrolyzate boils 10min, with enzyme-deactivating.Add 95% precipitation with alcohol, leave standstill, centrifugal, 55 ℃ of vacuum drying promptly get brown high-immunity regulation activity enzymolysis LBP-X 57.1g.
Embodiment three: with 250 gram matrimony vine schlempe oven dry, wear into powder just, add 1.75L water, 90 ℃ are extracted 3 times, each 2h, merging filtrate.60 ℃ are evaporated to original volume 1/10, add 95% ethanol in concentrate, make pure content reach 80%, leave standstill 15h, separate out precipitation.Sediment is in 60 ℃ of vacuum drying, and gained matrimony vine just polysaccharide adds that 350mL is water-soluble to be separated, and adds 70mL concentration and be 0.01% carbohydrase liquid, in 35 ℃ of hydrolysis 15min.Hydrolyzate boils 10min, with enzyme-deactivating.Add 95% precipitation with alcohol, leave standstill, centrifugal, 40 ℃ of vacuum drying promptly get brown high-immunity regulation activity enzymolysis LBP-X 48.8g.
Claims (2)
1, a kind of is the enzymolysis process of feedstock production high-immunity regulation activity LBP-X with the matrimony vine schlempe, it is characterized in that technical process is:
(1) with the oven dry of matrimony vine schlempe, wear into powder just, standby;
(2) add 6～9 times of water gagings, extract each 1～3h, filtration 1～3 time in 70～90 ℃;
(3) merge whole filtrates, 55～75 ℃ to be evaporated to original volume 1/10 standby;
(4) in concentrate, add 95% ethanol, make pure content reach 80%, leave standstill and separated out precipitation in 10～18 hours, get pure hypostasis and get pale brown look to the first polysaccharide of dark-brown matrimony vine, its LBP-X content 60～65%, weight average molecular weight 1～40,000 in 40～60 ℃ of vacuum drying;
(5) get matrimony vine polysaccharide just, be dissolved in water, be mixed with LBP-X concentration and be 2.4～3.6% first polysaccharide solution;
(6) get polysaccharide solution just, add the carbohydrase liquid of 1/5 volume, its concentration is 0.004～0.01%, and in 30～50 ℃ of hydrolysis 15～40min, hydrolyzate boils 10min, with enzyme-deactivating;
(7) adding 95% ethanol concentration of alcohol to the solution to hydrolyzate is 80%, produces precipitation, leaves standstill the centrifugal 15min of 3000r/min, taking precipitate 10～18 hours;
(8) sediment promptly gets its weight average molecular weight 1～30,000 of brown high-immunity regulation activity enzymolysis LBP-X in 40～60 ℃ of vacuum drying.
2, according to claim 1 is the enzymolysis process of feedstock production high-immunity regulation activity LBP-X with the matrimony vine schlempe, it is characterized in that raw materials for production also can directly use the fruit of Chinese wolfberry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100524007A CN100562525C (en) | 2007-06-07 | 2007-06-07 | A kind of is the enzymolysis process of feedstock production high-immunity regulation activity lycium barbarum polysaccharide with the matrimony vine schlempe |
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2007100524007A CN100562525C (en) | 2007-06-07 | 2007-06-07 | A kind of is the enzymolysis process of feedstock production high-immunity regulation activity lycium barbarum polysaccharide with the matrimony vine schlempe |
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| CN101069562A true CN101069562A (en) | 2007-11-14 |
| CN100562525C CN100562525C (en) | 2009-11-25 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103172759A (en) * | 2013-03-18 | 2013-06-26 | 中国科学院过程工程研究所 | Method for preparing functional polysaccharide and oligosaccharide with low molecular weights by using lyceum-barbarum polysaccharide |
| CN103833863A (en) * | 2012-11-20 | 2014-06-04 | 中国科学院兰州化学物理研究所 | Technology for preparing crude polysaccharides by extracting polysaccharides from Enteromorpha prolifera |
| WO2015010497A1 (en) * | 2013-07-23 | 2015-01-29 | 四川圣湖生物科技有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN105519828A (en) * | 2014-10-22 | 2016-04-27 | 天津芸熙生物技术有限公司 | Acidophile/grape ferment and application thereof |
| CN107853529A (en) * | 2017-12-07 | 2018-03-30 | 王春晓 | Mulberries particle Instant Drinks |
| CN108420826A (en) * | 2018-06-14 | 2018-08-21 | 吉林大学 | Polysaccharides is in the purposes prepared in treating or alleviating the drug of depression and includes its pharmaceutical composition |
| CN109055082A (en) * | 2018-06-29 | 2018-12-21 | 广西浙缘农业科技有限公司 | A kind of processing method of sugarcane |
| CN110128563A (en) * | 2019-06-06 | 2019-08-16 | 森田药妆股份有限公司 | A kind of ferment hydrolysis pholiota nameko polysaccharide and its preparation method and application |
-
2007
- 2007-06-07 CN CNB2007100524007A patent/CN100562525C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103833863A (en) * | 2012-11-20 | 2014-06-04 | 中国科学院兰州化学物理研究所 | Technology for preparing crude polysaccharides by extracting polysaccharides from Enteromorpha prolifera |
| CN103172759A (en) * | 2013-03-18 | 2013-06-26 | 中国科学院过程工程研究所 | Method for preparing functional polysaccharide and oligosaccharide with low molecular weights by using lyceum-barbarum polysaccharide |
| WO2015010497A1 (en) * | 2013-07-23 | 2015-01-29 | 四川圣湖生物科技有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN105519828A (en) * | 2014-10-22 | 2016-04-27 | 天津芸熙生物技术有限公司 | Acidophile/grape ferment and application thereof |
| CN107853529A (en) * | 2017-12-07 | 2018-03-30 | 王春晓 | Mulberries particle Instant Drinks |
| CN108420826A (en) * | 2018-06-14 | 2018-08-21 | 吉林大学 | Polysaccharides is in the purposes prepared in treating or alleviating the drug of depression and includes its pharmaceutical composition |
| CN108420826B (en) * | 2018-06-14 | 2020-03-10 | 吉林大学 | Application of lycium barbarum polysaccharide in preparation of medicine for treating or relieving depression and pharmaceutical composition containing lycium barbarum polysaccharide |
| CN109055082A (en) * | 2018-06-29 | 2018-12-21 | 广西浙缘农业科技有限公司 | A kind of processing method of sugarcane |
| CN110128563A (en) * | 2019-06-06 | 2019-08-16 | 森田药妆股份有限公司 | A kind of ferment hydrolysis pholiota nameko polysaccharide and its preparation method and application |
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|---|---|
| CN100562525C (en) | 2009-11-25 |
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