CN114350724A - Method for preparing garlic oligosaccharide through enzymolysis - Google Patents
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- 235000004611 garlic Nutrition 0.000 title claims abstract description 72
- 229920001542 oligosaccharide Polymers 0.000 title claims abstract description 39
- 150000002482 oligosaccharides Chemical class 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 28
- 244000245420 ail Species 0.000 title 1
- 240000002234 Allium sativum Species 0.000 claims abstract description 71
- 150000004676 glycans Chemical class 0.000 claims abstract description 37
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 37
- 239000005017 polysaccharide Substances 0.000 claims abstract description 37
- 239000002994 raw material Substances 0.000 claims abstract description 14
- 238000007710 freezing Methods 0.000 claims abstract description 9
- 230000008014 freezing Effects 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000000843 powder Substances 0.000 claims description 18
- 102000004190 Enzymes Human genes 0.000 claims description 12
- 108090000790 Enzymes Proteins 0.000 claims description 12
- 239000002244 precipitate Substances 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 10
- 239000003960 organic solvent Substances 0.000 claims description 10
- 229920001277 pectin Polymers 0.000 claims description 10
- 239000001814 pectin Substances 0.000 claims description 10
- 235000010987 pectin Nutrition 0.000 claims description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 230000000694 effects Effects 0.000 claims description 9
- 238000000227 grinding Methods 0.000 claims description 9
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 239000001110 calcium chloride Substances 0.000 claims description 7
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 7
- 238000002386 leaching Methods 0.000 claims description 7
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 238000002390 rotary evaporation Methods 0.000 claims description 6
- 238000003809 water extraction Methods 0.000 claims description 6
- 238000000746 purification Methods 0.000 claims description 5
- 238000005138 cryopreservation Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 230000007071 enzymatic hydrolysis Effects 0.000 claims 2
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 239000013543 active substance Substances 0.000 abstract description 6
- 230000002950 deficient Effects 0.000 abstract description 5
- 241000196324 Embryophyta Species 0.000 abstract description 4
- 238000000605 extraction Methods 0.000 abstract description 4
- 238000002360 preparation method Methods 0.000 abstract description 4
- 239000002699 waste material Substances 0.000 abstract description 2
- 206010040844 Skin exfoliation Diseases 0.000 description 13
- 229940029982 garlic powder Drugs 0.000 description 9
- 238000009835 boiling Methods 0.000 description 5
- 238000004140 cleaning Methods 0.000 description 5
- 238000005485 electric heating Methods 0.000 description 5
- 239000004570 mortar (masonry) Substances 0.000 description 5
- 238000000643 oven drying Methods 0.000 description 5
- 238000004321 preservation Methods 0.000 description 5
- 238000010298 pulverizing process Methods 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 230000003544 deproteinization Effects 0.000 description 4
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 238000005057 refrigeration Methods 0.000 description 3
- ZFTFOHBYVDOAMH-XNOIKFDKSA-N (2r,3s,4s,5r)-5-[[(2r,3s,4s,5r)-5-[[(2r,3s,4s,5r)-3,4-dihydroxy-2,5-bis(hydroxymethyl)oxolan-2-yl]oxymethyl]-3,4-dihydroxy-2-(hydroxymethyl)oxolan-2-yl]oxymethyl]-2-(hydroxymethyl)oxolane-2,3,4-triol Chemical class O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@@H]1[C@@H](O)[C@H](O)[C@](CO)(OC[C@@H]2[C@H]([C@H](O)[C@@](O)(CO)O2)O)O1 ZFTFOHBYVDOAMH-XNOIKFDKSA-N 0.000 description 2
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 2
- 229920002670 Fructan Polymers 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- -1 β -fructofuranosyl Chemical group 0.000 description 2
- 240000004246 Agave americana Species 0.000 description 1
- JDLKFOPOAOFWQN-VIFPVBQESA-N Allicin Natural products C=CCS[S@](=O)CC=C JDLKFOPOAOFWQN-VIFPVBQESA-N 0.000 description 1
- XUHLIQGRKRUKPH-GCXOYZPQSA-N Alliin Natural products N[C@H](C[S@@](=O)CC=C)C(O)=O XUHLIQGRKRUKPH-GCXOYZPQSA-N 0.000 description 1
- 244000298479 Cichorium intybus Species 0.000 description 1
- 235000007542 Cichorium intybus Nutrition 0.000 description 1
- XUHLIQGRKRUKPH-UHFFFAOYSA-N S-allyl-L-cysteine sulfoxide Natural products OC(=O)C(N)CS(=O)CC=C XUHLIQGRKRUKPH-UHFFFAOYSA-N 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 235000010081 allicin Nutrition 0.000 description 1
- JDLKFOPOAOFWQN-UHFFFAOYSA-N allicin Chemical compound C=CCSS(=O)CC=C JDLKFOPOAOFWQN-UHFFFAOYSA-N 0.000 description 1
- XUHLIQGRKRUKPH-DYEAUMGKSA-N alliin Chemical compound OC(=O)[C@@H](N)C[S@@](=O)CC=C XUHLIQGRKRUKPH-DYEAUMGKSA-N 0.000 description 1
- 235000015295 alliin Nutrition 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
Abstract
The invention belongs to the field of extraction of plant active substances, and particularly relates to a method for preparing garlic oligosaccharide by enzymolysis. Through the raw material and treatment, the crude polysaccharide is extracted and purified, and then the oligosaccharide is extracted by the garlic polysaccharide which is subjected to enzymolysis by endo-inulinase, so that the raw materials and the preparation method of the oligosaccharide are enriched, the additional value of garlic is improved, and the development of the garlic industry is driven. The method can also be used for extracting oligosaccharide from defective garlic, broken garlic or garlic damaged by freezing, thereby indirectly solving the problem that the garlic with lower quality is not easy to be reused and reducing unnecessary waste.
Description
Technical Field
The invention belongs to the field of extraction of plant active substances, and particularly relates to a method for preparing garlic oligosaccharide by enzymolysis.
Background
The extraction and separation technology of garlic active substances and the raw material resource of medicinal plants are key factors for restricting the industrialization of the garlic active substances. On one hand, the garlic has low content of active substances such as allicin, alliin and the like, so that the extraction and separation are difficult; on the other hand, although the garlic yield is large in China, in recent years, the price fluctuation of garlic markets is large, and the cost of raw materials for extracting medicinal components by simply taking garlic as a raw material is high, so that the profit of enterprises engaged in extracting active substances such as natural garlicin is slight.
Fructans are a class of polymers with linear or branched structures that have one or more β -fructofuranosyl linkages, which are chemical extensions of sucrose. Plants are the main source of fructans, such as chicory, agave, garlic, etc. The carbohydrate content in garlic is as high as 75% (dry basis), which is a good source for preparing garlic oligosaccharides.
The oligosaccharide has effects of regulating blood sugar, lowering blood pressure and blood lipid, promoting mineral absorption, regulating flora balance in vivo, and enhancing immunity. The method for preparing garlic oligosaccharide at the present stage is acidolysis, and the enzymolysis method for preparing garlic oligosaccharide is less. At present, most of the development of carbohydrates in garlic is limited to the development of garlic polysaccharide, and the preparation method of oligosaccharide is single.
Disclosure of Invention
Aiming at the problems existing in the prior stage, the invention provides a method for preparing garlic oligosaccharide by enzymolysis, which comprises the steps of extracting and purifying crude polysaccharide through raw material and treatment, and then extracting oligosaccharide through enzymolysis of garlic polysaccharide by endo-inulinase, so that the raw materials and the preparation method of oligosaccharide are enriched, the additional value of garlic is improved, and the development of garlic industry is driven.
The technical scheme of the invention is as follows:
a method for preparing garlic oligosaccharide by enzymolysis comprises the following steps:
(1) pretreatment of raw materials: peeling garlic; slicing and washing with clear water until no hand sticks; drying, powdering and storing for later use;
(2) extracting crude polysaccharide: extracting crude polysaccharide from Bulbus Allii by hot water extraction; cooling to room temperature, filtering, and leaching the filtered Bulbus Allii residue with hot water for 1-3 times;
(3) polysaccharide purification: adding CaCl2Removing pectin in the crude polysaccharide liquid obtained in the step (2) by using the solution, and repeating the operation until no precipitate exists; filtering, and concentrating the filtrate; adding sevag reagent to remove protein in the crude polysaccharide solution; removing the organic solvent by rotary evaporation; freezingDrying and grinding into powder; freezing and storing;
(4) enzymolysis of garlic polysaccharide: the enzyme activity is more than or equal to 2000U/g.
Preferably, the process of removing the protein in the crude sugar solution by the sevag reagent in the step (3) is as follows: adding the raw sugar solution into the mixture according to the volume ratio of 20 percent of the volume of the raw sugar solution: 1, and centrifuging to remove the protein until no white precipitate is produced.
Preferably, the hot water leaching process of the step (2) is as follows: according to the following steps of 1: leaching with hot water at 70-90 deg.C for 1-5 hr at a ratio of 10-20.
Preferably, the temperature for cryopreservation in step (3) is below-18 ℃.
Preferably, the step (3) is CaCl2The mass concentration of the solution is 8-12%.
The invention has the advantages of
Provides a method for quickly and efficiently preparing garlic oligosaccharide, and makes the preparation method of garlic oligosaccharide more diversified. The method can also be used for extracting oligosaccharide from defective garlic, broken garlic or garlic damaged by freezing, thereby indirectly solving the problem that the garlic with lower quality is not easy to be reused and reducing unnecessary waste.
The molecular weights of oligosaccharide with Degree of Polymerization (DP) of 3-5 are 504, 666 and 828 respectively, the total amount of oligosaccharide with DP less than or equal to 5 is used as determination index, DIONEX ICS-5000 is adopted＋And (3) measuring the content of oligosaccharide in the sample after the garlic polysaccharide is subjected to enzymolysis by using an ion chromatograph. And intercepting the oligosaccharide with the molecular weight of 300-1000Da by using a flow analyzer (ultrafiltration) so as to achieve the purpose of separation and purification, wherein the yield of the oligosaccharide is more than 80 percent.
Detailed Description
Example 1
A method for preparing garlic oligosaccharide by enzymolysis is prepared by the following steps:
pretreatment of raw materials: carrying out manual peeling or peeling treatment on the existing garlic by using a peeling machine;
preparing garlic powder: slicing peeled Bulbus Allii into 2-3mm slices, cleaning in flowing water until it is not sticky, oven drying at 65 deg.C, pulverizing into powder with pulverizer, and storing at 4 deg.C;
extracting garlic polysaccharide: taking garlic powder refrigerated at 4 ℃, and mixing the garlic powder according to the weight ratio of 1: 15 in a boiling pot or an electric heating jacketed pot filled with hot water of 80 ℃ for 2.5 h, cooling to room temperature, filtering with eight layers of sterile gauze, and repeating the above operation on the filtered garlic residue.
Removing pectin: adding 10% of CaCl2The solution was freed from pectin in the crude sugar solution and the procedure was repeated until no more precipitate was obtained. The crude sugar solution was concentrated using a rotary evaporator at a rotary evaporation temperature of 50 ℃.
sevag reagent deproteinization and organic solvent: adding the raw sugar solution into the mixture according to the volume ratio of 20 percent of the volume of the raw sugar solution: 1, centrifuging to remove proteins until no white precipitate is generated, separating by using a separating funnel, and removing the organic solvent in the crude sugar solution by using a rotary evaporator again.
Preservation of garlic polysaccharide: freeze drying, grinding into powder in mortar, and freezing at-18 deg.C;
enzymolysis of garlic polysaccharide: the enzymolysis temperature is 60 ℃, the enzyme addition amount is 25U/g (endo-inulinase, the enzyme activity is 2000U/g, the pH value is 5.0), the enzymolysis time is 6h, and the substrate concentration of enzymolysis is 5%.
Example 2
A method for preparing garlic oligosaccharide by enzymolysis is prepared by the following steps:
pretreatment of raw materials: carrying out manual peeling or peeling treatment on the existing garlic by using a peeling machine;
preparing garlic powder: slicing peeled Bulbus Allii into 2-3mm slices, cleaning in flowing water until it is not sticky, oven drying at 65 deg.C, pulverizing into powder with pulverizer, and storing at 4 deg.C;
extracting garlic polysaccharide: taking garlic powder refrigerated at 4 ℃, and mixing the garlic powder according to the weight ratio of 1: leaching 20 materials with water in a boiling pot or an electric heating jacketed pot filled with 70 deg.C hot water for 5 hr, cooling to room temperature, filtering with eight layers of sterile gauze, and repeating the above steps.
Removing pectin: adding 8% of CaCl2The solution was freed from pectin in the crude sugar solution and the procedure was repeated until no more precipitate was obtained. The crude sugar solution was concentrated using a rotary evaporator at a rotary evaporation temperature of 50 ℃.
sevag reagent deproteinization and organic solvent: the sugar solution is placed in a 500 ml separating funnel in batches, and 20% of the volume of the crude sugar solution is added into the mixture according to the volume ratio of 4: 1, centrifuging to remove proteins until no white precipitate is generated, and separating by using a separating funnel; and removing the organic solvent in the crude sugar solution by using a rotary evaporator again.
Preservation of garlic polysaccharide: freeze drying, grinding into powder in mortar, and freezing at-18 deg.C;
enzymolysis of garlic polysaccharide: the enzymolysis temperature is 60 ℃, the enzyme addition amount is 25U/g (endo-inulinase, the enzyme activity is 2000U/g, the pH value is 5.0), the enzymolysis time is 6h, and the substrate concentration of enzymolysis is 5%.
Example 3
A method for preparing garlic oligosaccharide by enzymolysis is prepared by the following steps:
pretreatment of raw materials: manually peeling or peeling off existing Bulbus Allii (fresh, defective, frozen injury, etc.);
preparing garlic powder: slicing peeled Bulbus Allii into 2-3mm slices, cleaning in flowing water until it is not sticky, oven drying at 65 deg.C, pulverizing into powder with pulverizer, and storing at 4 deg.C;
extracting garlic polysaccharide: extracting crude polysaccharide from Bulbus Allii by hot water extraction, collecting Bulbus Allii powder at 4 deg.C under refrigeration, and processing into powder according to the ratio of 1: leaching 10 materials with water in a boiling pot or an electric heating jacketed pot filled with 90 deg.C hot water for 1 hr, cooling to room temperature, filtering with eight layers of sterile gauze, and repeating the above steps.
Removing pectin: adding 12% of CaCl2Removing pectin from crude sugar solutionThe procedure was repeated until no more precipitate was present. The crude sugar solution was concentrated using a rotary evaporator at a rotary evaporation temperature of 50 ℃.
sevag reagent deproteinization and organic solvent: adding the raw sugar solution into the mixture according to the volume ratio of 20 percent of the volume of the raw sugar solution: 1, centrifuging to remove protein until no white precipitate is generated; filtering and separating; and removing the organic solvent in the crude sugar solution by using a rotary evaporator again.
Preservation of garlic polysaccharide: freeze drying, grinding into powder in mortar, and freezing at-18 deg.C;
enzymolysis of garlic polysaccharide: the enzymolysis temperature is 60 ℃, the enzyme addition amount is 25U/g (endo-inulinase, the enzyme activity is 2000U/g, the pH value is 5.0), the enzymolysis time is 6h, and the substrate concentration of enzymolysis is 5%.
Comparative example 1
A method for preparing garlic oligosaccharide by enzymolysis is prepared by the following steps:
pretreatment of raw materials: manually peeling or peeling off existing Bulbus Allii (fresh, defective, frozen injury, etc.);
preparing garlic powder: slicing peeled Bulbus Allii into 2-3mm slices, cleaning in flowing water until it is not sticky, oven drying at 65 deg.C, pulverizing into powder with pulverizer, and storing at 4 deg.C;
extracting garlic polysaccharide: extracting crude polysaccharide from Bulbus Allii by hot water extraction, collecting Bulbus Allii powder at 4 deg.C under refrigeration, and processing into powder according to the ratio of 1: 15 in a boiling pot or an electric heating jacketed pot filled with hot water of 80 ℃ for 2.5 h, cooling to room temperature, filtering with eight layers of sterile gauze, and repeating the above operation on the filtered garlic residue.
sevag reagent deproteinization and organic solvent: the sugar solution is placed in a 500 mL separating funnel in batches, and 20% of the volume of the crude sugar solution is added into the mixture according to the volume ratio of 4: 1, and centrifuging to remove the protein until no white precipitate is produced. And removing the organic solvent in the crude sugar solution by using a rotary evaporator again.
Preservation of garlic polysaccharide: freeze drying, grinding into powder in mortar, and freezing at-18 deg.C;
enzymolysis of garlic polysaccharide: the enzymolysis temperature is 60 ℃, the enzyme addition amount is 25U/g (endo-inulinase, the enzyme activity is 2000U/g, the pH value is 5.0), the enzymolysis time is 6h, and the substrate concentration of enzymolysis is 5%.
Comparative example 2
A method for preparing garlic oligosaccharide by enzymolysis is prepared by the following steps:
pretreatment of raw materials: manually peeling or peeling off existing Bulbus Allii (fresh, defective, frozen injury, etc.);
preparing garlic powder: slicing peeled Bulbus Allii into 2-3mm slices, cleaning in flowing water until it is not sticky, oven drying at 65 deg.C, pulverizing into powder with pulverizer, and storing at 4 deg.C;
extracting garlic polysaccharide: extracting crude polysaccharide from Bulbus Allii by hot water extraction, collecting Bulbus Allii powder at 4 deg.C under refrigeration, and processing into powder according to the ratio of 1: 15 in a boiling pot or an electric heating jacketed pot filled with hot water of 80 ℃ for 2.5 h, cooling to room temperature, filtering with eight layers of sterile gauze, and repeating the above operation on the filtered garlic residue.
Removing pectin: adding 10% of CaCl2The solution was freed from pectin in the crude sugar solution and the procedure was repeated until no more precipitate was obtained. The crude sugar solution was concentrated using a rotary evaporator at a rotary evaporation temperature of 50 ℃.
Preservation of garlic polysaccharide: freeze drying, grinding into powder in mortar, and freezing at-18 deg.C;
enzymolysis of garlic polysaccharide: the enzymolysis temperature is 60 ℃, the enzyme addition amount is 25U/g (endo-inulinase, the enzyme activity is 2000U/g, the pH value is 5.0), the enzymolysis time is 6h, and the substrate concentration of enzymolysis is 5%.
Examples of the effects of the invention
And (3) separation and purification of oligosaccharide: the molecular weights of oligosaccharide with Degree of Polymerization (DP) of 3-5 are 504, 666 and 828 respectively, the total amount of oligosaccharide with DP less than or equal to 5 is used as determination index, DIONEX ICS-5000 is adopted＋Determination of garlic by ion chromatographyAnd (3) the polysaccharide is subjected to enzymolysis, and the content of oligosaccharide in the sample is low. And intercepting oligosaccharide with molecular weight of 300-1000Da by using a flow analyzer (ultrafiltration) to achieve the purpose of separation and purification, 500g of garlic is respectively adopted to extract according to the methods of examples 1-3 and comparative examples 1 and 2, and the specific data are shown in Table 1.
Claims (5)
1. A method for preparing garlic oligosaccharide by enzymolysis is characterized by comprising the following steps:
(1) pretreatment of raw materials: peeling garlic; slicing and washing with clear water until no hand sticks; drying, powdering and storing for later use;
(2) extracting crude polysaccharide: extracting crude polysaccharide from Bulbus Allii by hot water extraction; cooling to room temperature, filtering, and leaching the filtered Bulbus Allii residue with hot water for 1-3 times;
(3) polysaccharide purification: adding CaCl2Removing pectin in the crude polysaccharide liquid obtained in the step (2) by using the solution, and repeating the operation until no precipitate exists; filtering, and concentrating the filtrate; adding sevag reagent to remove protein in the crude polysaccharide solution; removing the organic solvent by rotary evaporation; freeze drying, and grinding into powder; freezing and storing;
(4) enzymolysis of garlic polysaccharide: the enzyme activity is more than or equal to 2000U/g.
2. The method for preparing garlic oligosaccharide through enzymolysis according to claim 1, wherein the step (3) of sevag reagent for removing protein in the crude sugar solution comprises the following steps: adding the raw sugar solution into the mixture according to the volume ratio of 20 percent of the volume of the raw sugar solution: 1, and centrifuging to remove the protein until no white precipitate is produced.
3. The method for preparing garlic oligosaccharide by enzymolysis as claimed in claim 1, wherein the hot water extraction process in step (2) is as follows: according to the following steps of 1: leaching with hot water at 70-90 deg.C for 1-5 hr at a ratio of 10-20.
4. The enzymatic hydrolysis method for preparing garlic oligosaccharide according to claim 1, wherein the temperature of the cryopreservation in the step (3) is lower than-18 ℃.
5. The enzymatic hydrolysis process for preparing garlic oligosaccharide as claimed in claim 1, wherein the CaCl in step (3) is added2The mass concentration of the solution is 8-12%.
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