CN109134676A - Oligosaccharide and preparation method thereof, application - Google Patents
Oligosaccharide and preparation method thereof, application Download PDFInfo
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- CN109134676A CN109134676A CN201810824628.1A CN201810824628A CN109134676A CN 109134676 A CN109134676 A CN 109134676A CN 201810824628 A CN201810824628 A CN 201810824628A CN 109134676 A CN109134676 A CN 109134676A
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- 229920001542 oligosaccharide Polymers 0.000 title claims abstract description 56
- 150000002482 oligosaccharides Chemical class 0.000 title claims abstract description 56
- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 239000012528 membrane Substances 0.000 claims abstract description 56
- 239000011347 resin Substances 0.000 claims abstract description 53
- 229920005989 resin Polymers 0.000 claims abstract description 53
- 150000004676 glycans Chemical class 0.000 claims abstract description 29
- 239000007788 liquid Substances 0.000 claims abstract description 29
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 29
- 239000005017 polysaccharide Substances 0.000 claims abstract description 29
- 239000012530 fluid Substances 0.000 claims abstract description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000003814 drug Substances 0.000 claims abstract description 21
- 239000000706 filtrate Substances 0.000 claims abstract description 21
- 238000001914 filtration Methods 0.000 claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000002699 waste material Substances 0.000 claims abstract description 18
- 238000001556 precipitation Methods 0.000 claims abstract description 16
- 108010059892 Cellulase Proteins 0.000 claims abstract description 14
- 229940106157 cellulase Drugs 0.000 claims abstract description 14
- 235000019441 ethanol Nutrition 0.000 claims abstract description 13
- 230000009514 concussion Effects 0.000 claims abstract description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 24
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 229940088598 enzyme Drugs 0.000 claims description 10
- 239000000047 product Substances 0.000 claims description 10
- 238000004108 freeze drying Methods 0.000 claims description 9
- 241000894006 Bacteria Species 0.000 claims description 7
- 229940079593 drug Drugs 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 5
- 230000036541 health Effects 0.000 claims description 5
- 230000009286 beneficial effect Effects 0.000 claims description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 claims description 4
- 239000000470 constituent Substances 0.000 claims description 2
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 claims description 2
- 229920001277 pectin Polymers 0.000 claims 1
- 235000010987 pectin Nutrition 0.000 claims 1
- 239000001814 pectin Substances 0.000 claims 1
- 238000012545 processing Methods 0.000 abstract description 34
- 239000006041 probiotic Substances 0.000 abstract description 14
- 235000018291 probiotics Nutrition 0.000 abstract description 14
- 230000035755 proliferation Effects 0.000 abstract description 10
- 239000002994 raw material Substances 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract description 2
- 210000004379 membrane Anatomy 0.000 description 33
- 238000000034 method Methods 0.000 description 17
- 238000001035 drying Methods 0.000 description 13
- 239000000243 solution Substances 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 12
- 230000007071 enzymatic hydrolysis Effects 0.000 description 11
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 11
- 239000000843 powder Substances 0.000 description 8
- 238000010790 dilution Methods 0.000 description 7
- 239000012895 dilution Substances 0.000 description 7
- 239000004744 fabric Substances 0.000 description 6
- 240000000249 Morus alba Species 0.000 description 5
- 235000008708 Morus alba Nutrition 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 230000000968 intestinal effect Effects 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 3
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 230000000996 additive effect Effects 0.000 description 3
- 230000009849 deactivation Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 229940039696 lactobacillus Drugs 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 150000002772 monosaccharides Chemical class 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 239000000049 pigment Substances 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000002893 slag Substances 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000001117 sulphuric acid Substances 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 235000021255 galacto-oligosaccharides Nutrition 0.000 description 2
- 150000003271 galactooligosaccharides Chemical class 0.000 description 2
- 239000011812 mixed powder Substances 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 244000170916 Paeonia officinalis Species 0.000 description 1
- 235000006484 Paeonia officinalis Nutrition 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 239000006286 aqueous extract Substances 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000007661 gastrointestinal function Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229940023492 oral liquid product Drugs 0.000 description 1
- 229940100688 oral solution Drugs 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 235000013406 prebiotics Nutrition 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 235000019722 synbiotics Nutrition 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 238000004260 weight control Methods 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7016—Disaccharides, e.g. lactose, lactulose
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Polymers & Plastics (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Nutrition Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Food Science & Technology (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Sustainable Development (AREA)
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- Materials Engineering (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention relates to a kind of oligosaccharide and preparation method thereof, application.The preparation method of the oligosaccharide includes: the waste liquid for taking Chinese medicine to generate in aqueous extraction-alcohol precipitation technology, and evaporative removal ethyl alcohol filters, obtains filtrate；UF membrane is carried out to the filtrate and obtains efflux, UF membrane is carried out to the efflux and obtains trapped fluid；Macroreticular resin decoloration, concussion are added into the trapped fluid, filtering removal macroreticular resin obtains polysaccharide solution；First into the polysaccharide solution be added cellulase digested, add pectase and digested, enzyme-deactivating to get.The waste liquid that the present invention is generated in water extract-alcohol precipitation processing using Chinese medicine is raw material, UF membrane twice first is carried out to waste liquid, macroreticular resin decoloration is carried out again, finally successively digested using cellulase and pectase, molecular weight can be obtained in 2000 oligosaccharide below, such oligosaccharide is easier to be utilized by probiotics, can play good facilitation to the proliferation of probiotics.
Description
Technical field
The present invention relates to oligosaccharide extractive technique fields, more particularly to oligosaccharide and preparation method thereof, application.
Background technique
Oligosaccharide is a kind of new type functional sugar source, is widely used in the neck such as food, health care product, medicine, feed addictive
Domain.Oligosaccharide can improve micro-ecological environment in human body, adjust gastrointestinal function, be conducive to the proliferation of probiotics and inhibit spoilage organisms
Growth；Meanwhile it has the function of promoting the development of Intestinal Mucosal Immunity system, also has promotion to parenterally systemic immune system
Effect；Oligosaccharide can also improve the vigor of intestinal mucosa cells, increase the suction of enteron aisle by the microecological balance of adjusting intestinal flora
Receive the ability of minerals.
The preparation process of traditional oligosaccharide, for example, preparing the work of oligosaccharide using the peony seeds dregs of rice, mulberry fruit juice as raw material
Skill, these traditional handicrafts obtain not strong containing oligosaccharide product functionality, very to the Effect of promoting growth of beneficial bacteria of intestinal tract
It is limited.
Traditional Chinese medicine enterprise generally requires to carry out water extracting alcohol to Chinese medicine when producing the products such as medicinal extract, granule and oral solution
Heavy processing, treated, and waste liquid is often directly thrown away, larger to environmental disruption.
Currently, the waste liquid for also not having the water extract-alcohol precipitation for using Chinese medicine to generate is raw material and produces proliferation of intestinal probiotics
The method of the apparent oligosaccharide of facilitation.
Summary of the invention
Based on this, prepared the main purpose of the present invention is to provide a kind of by raw material of the waste liquid of the water extract-alcohol precipitation of Chinese medicine
To the method for the apparent oligosaccharide of proliferation of intestinal probiotics facilitation.
The purpose of the present invention is what is be achieved through the following technical solutions:
A kind of preparation method of oligosaccharide, the preparation method include:
Step 1: the waste liquid for taking Chinese medicine to generate in aqueous extraction-alcohol precipitation technology, evaporative removal ethyl alcohol, filtering obtain filtrate；
Step 2: carrying out UF membrane to the filtrate obtains efflux, UF membrane is carried out to the efflux and obtains trapped fluid；
Step 3: macroreticular resin decoloration is added into the trapped fluid, it is molten to obtain polysaccharide for concussion, filtering removal macroreticular resin
Liquid；
It is digested Step 4: cellulase is first added into the polysaccharide solution, adds pectase and digested, enzyme
Inactivation to get.
In wherein some embodiments, to the filtrate carry out UF membrane when use molecular cut off for 250kDa~
The ultrafiltration membrane of 300kDa；Use molecular cut off for the ultrafiltration membrane of 3kDa~5kDa when carrying out UF membrane to efflux；It is described big
Hole resin is in AB-8 resin, D101 resin, HPD-300 resin, X-5 resin, NK-2 resin, NKA-2 resin, NK-9 resin
At least one, the quality that macroreticular resin is added in every milliliter of trapped fluid is 0.18g~0.22g；The addition fiber
The condition that plain enzyme is digested are as follows: 30 DEG C~50 DEG C, 2h~5h；The condition that pectase is digested is added are as follows: 30 DEG C~50 DEG C,
2h~5h.
In wherein some embodiments, to the filtrate carry out UF membrane when use molecular cut off for 280kDa~
The ultrafiltration membrane of 300kDa；Use molecular cut off for the ultrafiltration membrane of 3kDa~5kDa when carrying out UF membrane to efflux；It is described big
Hole resin is selected from AB-8 resin, and the quality that macroreticular resin is added in every milliliter of trapped fluid is 0.20g~0.22g；Described
The condition that cellulase is digested is added are as follows: 30 DEG C~50 DEG C, 2h~5h；The condition that pectase is digested is added are as follows: 30
DEG C~50 DEG C, 2h~5h.
In wherein some embodiments, use molecular cut off for the ultrafiltration of 300kDa when carrying out UF membrane to the filtrate
Film；Use molecular cut off for the ultrafiltration membrane of 5kDa when carrying out UF membrane to efflux.
In wherein some embodiments, the macroreticular resin is AB-8 resin, and macropore is added in every milliliter of trapped fluid
The quality of resin is 0.2g.
In wherein some embodiments, step 4 includes the steps that being freeze-dried the product after the enzyme-deactivating.
In wherein some embodiments, the condition of the freeze-drying are as follows: -0.05MPa~-0.08MPa, -45 DEG C~-35
DEG C, 2.5d~3d.
It is a further object of the present invention to provide a kind of oligosaccharide that above-mentioned preparation method obtains.
It is yet another object of the invention to provide a kind of above-mentioned oligosaccharide preparation can improve beneficial bacteria of intestinal tract food,
Application in health care product or drug.
Another object of the present invention be to provide oligosaccharide that more than one are stated be active constituent can to improve enteron aisle prebiotic
Food, health care product or the drug of bacterium.
Compared with prior art, the present invention have it is following the utility model has the advantages that
The waste liquid that the present invention is generated in water extract-alcohol precipitation processing using Chinese medicine for the first time first carries out film twice to waste liquid as raw material
Separation, then macroreticular resin decoloration is carried out, it is finally successively digested using cellulase and pectase, is based particularly on Chinese medicine
Waste liquid this raw material generated in water extract-alcohol precipitation processing, corresponding cooperation is using suitable ultrafiltration membrane, macroreticular resin and enzyme
Solution condition forms specific oligosaccharide preparation process, when carrying out oligosaccharide preparation by the preparation process, can be good at extracting
Molecular weight ranges contained in the waste liquid of Chinese medicine water extract-alcohol precipitation processing are ten tens of thousands of polysaccharide to hundreds of thousands, and are broken down into
In 2000 oligosaccharide below, such oligosaccharide is easier to be utilized by probiotics for molecular weight control, can be to probiotics
Proliferation plays good facilitation.Meanwhile the preparation process is successfully realized the recycling of traditional Chinese medicine enterprise waste liquid, increase through
Ji benefit, and be conducive to protect environment.
Specific embodiment
It to facilitate the understanding of the present invention, below will be to invention is more fully described.But the present invention can be to be permitted
Mostly different form is realized, however it is not limited to embodiment described herein.On the contrary, purpose of providing these embodiments is makes
It is more thorough and comprehensive to the understanding of the disclosure.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical.Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.Term as used herein "and/or" includes one or more phases
Any and all combinations of the listed item of pass.
Water extraction and alcohol precipitation method namely water alcohol method known in water extract-alcohol precipitation, that is, field of traditional Chinese medicine extraction of the present invention, refer in
In medicine water extracting liquid, ethyl alcohol, which is added, to be made up to different alcohol contents, and some drugs ingredient solubility in alcoholic solution, which reduces to be precipitated, sinks
It forms sediment, enables the method that Aqueous extracts refine after separation of solid and liquid.The Chinese medicine that the present invention mentions carries out the waste liquid of water extract-alcohol precipitation generation,
Refer specifically to gained liquid portion after above-mentioned separation of solid and liquid.
Embodiment 1
This example provides a kind of oligosaccharide and preparation method thereof.Whole preparation process is as follows:
It takes and is to the waste liquid after mulberry branch progress water extract-alcohol precipitation → revolving volatilization ethyl alcohol → filtering → molecular cut off
UF membrane → efflux → molecular cut off of 300kDa is UF membrane → trapped fluid → macroreticular resin decoloration → drying of 5kDa
→ survey polysaccharide concentration → dilution → enzymatic hydrolysis → enzyme-deactivating → freeze-drying；Wherein:
Filtering: extracting solution first uses the filter-cloth filtering of 300 mesh, then with the filter-cloth filtering of 600 mesh, takes filtrate, filter residue；Again toward filter
The pure water of 10 times of volumes is added in slag, extracts again according to the above method one time；Merge filtrate obtained as above after filtering.
UF membrane: it is 300kDa Ultra filtration membrane that filtrate, which first passes through molecular cut off, to remove most of protein, until
Stopped in phend-sulphuric acid measurement efflux without polysaccharide, takes efflux；Then passing through molecular cut off again is 5kDa ultrafiltration
UF membrane, until efflux takes trapped fluid in colourless stopping removing the small-molecule substances such as monosaccharide, pigment and inorganic salts；Film point
From the pressure used is operated as 0.3MPa, temperature is room temperature, membrane area 0.2m2。
Macroreticular resin decoloration: the macropore pre-processed is added in resin/polysaccharide solution (0.2g/mL) ratio into trapped fluid
Resin A B-8, then places it in constant temperature oscillator, and (120r/min) is vibrated at 37 DEG C and adsorbs 5h, is filtered to remove macropore tree
Rouge, the polysaccharide solution after being decolourized.
Drying: 60 DEG C of drying 5h, until blocky.
Survey polysaccharide concentration: with block-like polysaccharide concentration after sulfuric acid-phynol method survey drying.
Dilution: polysaccharide concentration is diluted to 50mg/mL.
Enzymatic hydrolysis: first using and digest 4h at 40 DEG C of cellulase, then uses and digest 4h at 40 DEG C of pectase.
Enzyme-deactivating: 4min is boiled into enzymolysis liquid heating.
Freeze-drying: liquid is freeze-dried 3d at -0.08MPa vacuum degree and -40 DEG C after enzyme deactivation, obtains functional oligomeric
Icing Sugar end.
Embodiment 2
This example provides a kind of oligosaccharide and preparation method thereof.Whole preparation process is as follows:
It takes and is to the waste liquid after mulberry branch progress water extract-alcohol precipitation → revolving volatilization ethyl alcohol → filtering → molecular cut off
UF membrane → efflux → molecular cut off of 250kDa is UF membrane → trapped fluid → macroreticular resin decoloration → drying of 3kDa
→ survey polysaccharide concentration → dilution → enzymatic hydrolysis → enzyme-deactivating → freeze-drying；Wherein:
Filtering: extracting solution first uses the filter-cloth filtering of 300 mesh, then with the filter-cloth filtering of 600 mesh, takes filtrate, filter residue；Again toward filter
The pure water of 10 times of volumes is added in slag, extracts again according to the above method one time；Merge filtrate obtained as above after filtering.
UF membrane: it is 250kDa Ultra filtration membrane that filtrate, which first passes through molecular cut off, to remove most of protein, until
Stopped in phend-sulphuric acid measurement efflux without polysaccharide, takes efflux；Then passing through molecular cut off again is 3kDa ultrafiltration
UF membrane, until efflux takes trapped fluid in colourless stopping removing the small-molecule substances such as monosaccharide, pigment and inorganic salts；Film point
From the pressure used is operated as 0.3MPa, temperature is room temperature, membrane area 0.2m2。
Macroreticular resin decoloration: into trapped fluid by resin/polysaccharide solution (0.18g/mL) ratio be added pre-processed it is big
Hole Resin A B-8, then places it in constant temperature oscillator, and (120r/min) is vibrated at 37 DEG C and adsorbs 5h, is filtered to remove macropore
Resin, the polysaccharide solution after being decolourized.
Drying: 60 DEG C of drying 4h, until blocky.
Survey polysaccharide concentration: with block-like polysaccharide concentration after sulfuric acid-phynol method survey drying.
Dilution: polysaccharide concentration is diluted to 1mg/mL.
Enzymatic hydrolysis: first using and digest 2h at 30 DEG C of cellulase, then uses and digest 2h at 30 DEG C of pectase.
Enzyme-deactivating: 3~5min is boiled into enzymolysis liquid heating.
Freeze-drying: liquid is freeze-dried 3d at -0.08MPa vacuum degree and -40 DEG C after enzyme deactivation, obtains functional oligomeric
Icing Sugar end.
Embodiment 3
This example provides a kind of oligosaccharide and preparation method thereof.Whole preparation process is as follows:
It takes and is to the waste liquid after mulberry branch progress water extract-alcohol precipitation → revolving volatilization ethyl alcohol → filtering → molecular cut off
UF membrane → efflux → molecular cut off of 280kDa is UF membrane → trapped fluid → macroreticular resin decoloration → drying of 4kDa
→ survey polysaccharide concentration → dilution → enzymatic hydrolysis → enzyme-deactivating → freeze-drying；Wherein:
Filtering: extracting solution first uses the filter-cloth filtering of 300 mesh, then with the filter-cloth filtering of 600 mesh, takes filtrate, filter residue；Again toward filter
The pure water of 10 times of volumes is added in slag, extracts again according to the above method one time；Merge filtrate obtained as above after filtering.
UF membrane: it is 280kDa Ultra filtration membrane that filtrate, which first passes through molecular cut off, to remove most of protein, until
Stopped in phend-sulphuric acid measurement efflux without polysaccharide, takes efflux；Then passing through molecular cut off again is 4kDa ultrafiltration
UF membrane, until efflux takes trapped fluid in colourless stopping removing the small-molecule substances such as monosaccharide, pigment and inorganic salts；Film point
From the pressure used is operated as 0.3MPa, temperature is room temperature, membrane area 0.2m2。
Macroreticular resin decoloration: into trapped fluid by resin/polysaccharide solution (0.22g/mL) ratio be added pre-processed it is big
Hole Resin A B-8, then places it in constant temperature oscillator, and (120r/min) is vibrated at 37 DEG C and adsorbs 5h, is filtered to remove macropore
Resin, the polysaccharide solution after being decolourized.
Drying: 60 DEG C of drying 6h, until blocky.
Survey polysaccharide concentration: with block-like polysaccharide concentration after sulfuric acid-phynol method survey drying.
Dilution: polysaccharide concentration is diluted to 100mg/mL.
Enzymatic hydrolysis: first using and digest 5h at 50 DEG C of cellulase, then uses and digest 5h at 50 DEG C of pectase.
Enzyme-deactivating: 5min is boiled into enzymolysis liquid heating.
Freeze-drying: liquid is freeze-dried 3d at -0.08MPa vacuum degree and -40 DEG C after enzyme deactivation, obtains functional oligomeric
Icing Sugar end.
Comparative example 1
This example is the comparative example of embodiment 1, and the main distinction with embodiment 1 is, technique is not first Ultra filtration membrane, again
Macroreticular resin decoloration finally digests, but first enzymatic hydrolysis, again Ultra filtration membrane, the decoloration of last macroreticular resin, and technique is as follows:
It takes and goes out to the waste liquid generated after mulberry branch progress water extract-alcohol precipitation → revolving volatilization ethyl alcohol → filtering → enzymatic hydrolysis → enzyme
UF membrane → trapped fluid that UF membrane → efflux → molecular cut off that work → molecular cut off is 300kDa is 5kDa → big
Hole resin decolorization → drying → survey polysaccharide concentration → dilution → freeze-drying.
Comparative example 2
This example is the comparative example of embodiment 1, from the main distinction of embodiment 1 be enzymatic hydrolysis the step of it is different: the example be not according to
It is secondary to be digested with cellulase and pectase, but cellulase and pectinase enzymatic hydrolysis are added simultaneously.
Comparative example 3
This example is the comparative example of embodiment 1, the main distinction with embodiment 1 are as follows:
(1) when UF membrane, first using 350kDa Ultra filtration membrane, again using 10kDa Ultra filtration membrane；
(2) it first used when digesting and digests 6h at 28 DEG C of cellulase, use pectase to digest 1.5h at 55 DEG C again.
Performance test
Example 1, embodiment 2, embodiment 3, comparative example 1, comparative example 2, comparative example 3, gained oligosaccharide respectively, and
It is commercialized oligosaccharide (galactooligosaccharide, oligoisomaltose, as positive control), MRS is added to the additive amount of 15% (w/v)
In culture medium, first inoculated plant lactobacillus is inoculated with Lactobacillus rhamnosus when OD value reaches 0.5.Culture counts clump count.
Wherein, the processing that the processing of 1 gained oligosaccharide of embodiment is denoted as processing 1, adds 2 gained oligosaccharide of embodiment is added
It is denoted as processing 2, adds the processing note that the processing of 3 gained oligosaccharide of embodiment is denoted as processing 3, adds 1 gained oligosaccharide of comparative example
The processing for dealing with 4, addition 2 gained oligosaccharide of comparative example is denoted as processing 5, the processing of addition 3 gained oligosaccharide of comparative example is denoted as
Processing 6, the processing of addition commercialization galactooligosaccharide are denoted as processing 7, the processing of addition commercialization oligoisomaltose is denoted as locating
8 (positive controls) of reason, the processing that oligosaccharide is not added are denoted as processing 9 (negative control).
According to the above table 1:
Processing 1 and processing 2, processing 3 can obtain more clump count, illustrate embodiment 1, embodiment 2, embodiment 3
Obtained oligosaccharide can play the significant effect for promoting proliferation of probiotics.But, it compares, what processing 1 obtained
Clump count is more, this explanation, oligosaccharide preparation process of the invention has preferably scheme.Processing 1 is relative to processing 6, bacterium colony
Number dramatically increases, this explanation, and the selection of suitable ultrafiltration membrane, macroreticular resin, enzymatic hydrolysis condition, is important in preparation process.
Processing 1 is relative to processing 4 and processing 5, and clump count dramatically increases, this illustrates the oligosaccharide that present invention process is prepared
Vital effect is played to the proliferation of probiotics, if be adjusted to technique, preparation-obtained product, although
It can promote the proliferation of probiotics, but it is very limited for promoting degree.
Compared with processing 9, the proliferation rate of probiotics reaches 619% for processing 1, compared with processing 7, processing 8, the increasing of probiotics
The rate of growing reaches 361% and 382%.This explanation, the oligosaccharide that embodiment 1 is prepared is relative to commercial goods oligosaccharide, more
It is easy to be utilized by probiotics, to be conducive to proliferation of probiotics.
Application examples 1
Oligosaccharide powder prepared by embodiment 1 is added in MRS culture medium by the additive amount of 10~20% (w/v), first
Inoculated plant lactobacillus is inoculated with Lactobacillus rhamnosus when OD value reaches 0.5, carries out initial concentration to thallus after cultivating 18h, and
Quick freeze carries out pellet little particle at pellet little particle, and after being freeze-dried by cryoprotectors such as addition soybean powders afterwards
Grinding remixes, to realize uniformity, finally packing is fabricated to individual packets dress.
Application examples 2
Oligosaccharide powder prepared by embodiment 2 is added in MRS culture medium by the additive amount of 10~20% (w/v), first
Inoculated plant lactobacillus is inoculated with Lactobacillus rhamnosus when OD value reaches 0.5, carries out initial concentration to thallus after cultivating 18h, and
Quick freeze grinds again bacterium powder at pellet little particle, and after being freeze-dried by cryoprotectors such as addition soybean powders afterwards
Mixing obtains mixed powder, and finally adding the oligosaccharide powder of embodiment 2 again, (mass ratio of mixed powder and oligosaccharide powder is
4:1) it is fabricated to synbiotic.
Application examples 3
After oligosaccharide powder prepared by embodiment 3 is dissolved in pure water by 40~60% (w/v), 5% (w/v) xylose is added
Alcohol, packing are fabricated to independent oral liquid product.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of preparation method of oligosaccharide, which is characterized in that the preparation method includes:
Step 1: taking the waste liquid for carrying out water extract-alcohol precipitation generation to Chinese medicine, evaporative removal ethyl alcohol, filtering obtains filtrate；
Step 2: carrying out UF membrane to the filtrate obtains efflux, UF membrane is carried out to the efflux and obtains trapped fluid；
Step 3: macroreticular resin decoloration is added into the trapped fluid, concussion, filtering removal macroreticular resin obtains polysaccharide solution；
It is digested Step 4: cellulase is first added into the polysaccharide solution, adds pectase and digested, enzyme goes out
It is living to get.
2. the preparation method of oligosaccharide according to claim 1, which is characterized in that adopted when carrying out UF membrane to the filtrate
The ultrafiltration membrane for being 250kDa~300kDa with molecular cut off；Use molecular cut off for 3kDa when carrying out UF membrane to efflux
The ultrafiltration membrane of~5kDa；The macroreticular resin is selected from AB-8 resin, D101 resin, HPD-300 resin, X-5 resin, NK-2 tree
At least one of rouge, NKA-2 resin, NK-9 resin, the quality that macroreticular resin is added in every milliliter of trapped fluid is 0.18g
~0.22g；The condition that the addition cellulase is digested are as follows: 30 DEG C~50 DEG C, 2h~5h；Pectase is added and carries out enzyme
The condition of solution are as follows: 30 DEG C~50 DEG C, 2h~5h.
3. the preparation method of oligosaccharide according to claim 2, which is characterized in that adopted when carrying out UF membrane to the filtrate
The ultrafiltration membrane for being 280kDa~300kDa with molecular cut off；Use molecular cut off for 3kDa when carrying out UF membrane to efflux
The ultrafiltration membrane of~5kDa；The macroreticular resin is selected from AB-8 resin, and the quality of macroreticular resin is added in every milliliter of trapped fluid
For 0.20g~0.22g；The condition that the addition cellulase is digested are as follows: 30 DEG C~50 DEG C, 2h~5h；Pectin is added
The condition that enzyme is digested are as follows: 30 DEG C~50 DEG C, 2h~5h.
4. the preparation method of oligosaccharide according to claim 3, which is characterized in that adopted when carrying out UF membrane to the filtrate
The ultrafiltration membrane for being 300kDa with molecular cut off；Use molecular cut off for the ultrafiltration of 5kDa when carrying out UF membrane to efflux
Film.
5. the preparation method of oligosaccharide according to claim 3, which is characterized in that the macroreticular resin is AB-8 resin,
The quality that macroreticular resin is added in every milliliter of trapped fluid is 0.2g.
6. the preparation method of oligosaccharide according to any one of claims 1 to 5, which is characterized in that step 4 includes to institute
The step of product after stating enzyme-deactivating is freeze-dried.
7. the preparation method of oligosaccharide according to claim 6, which is characterized in that the condition of the freeze-drying are as follows:-
0.05MPa~-0.08MPa, -45 DEG C~-35 DEG C, 2.5d~3d.
8. the oligosaccharide that the described in any item preparation methods of claim 1 to 7 obtain.
9. oligosaccharide according to any one of claims 8 answering in the food, health care product or drug that preparation can improve beneficial bacteria of intestinal tract
With.
10. a kind of using oligosaccharide according to any one of claims 8 as the food that can improve beneficial bacteria of intestinal tract, the health care product of active constituent
Or drug.
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| CN113143825A (en) * | 2021-05-10 | 2021-07-23 | 广东省农业科学院蚕业与农产品加工研究所 | A collutory containing Chinese medicinal prebiotics and its preparation method |
| CN114916620A (en) * | 2022-05-23 | 2022-08-19 | 广西五和博澳药业有限公司 | Ramulus mori oligosaccharide prebiotics cattle feed and preparation method and application thereof |
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Effective date of registration: 20210517 Address after: No.16 Jincheng Road, Qingyuan Town, Yizhou District, Hechi City, Guangxi Zhuang Autonomous Region 546300 Patentee after: Guangxi Wuhe Boao Pharmaceutical Co.,Ltd. Address before: No. 133, Yiheng Road, Dongguan Zhuang, Tianhe District, Guangzhou, Guangdong 510665 Patentee before: SERICULTURE & AGRI FOOD Research Institute GAAS |