CN106117393A - A kind of method preparing oligochitosan - Google Patents
A kind of method preparing oligochitosan Download PDFInfo
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- CN106117393A CN106117393A CN201610848221.3A CN201610848221A CN106117393A CN 106117393 A CN106117393 A CN 106117393A CN 201610848221 A CN201610848221 A CN 201610848221A CN 106117393 A CN106117393 A CN 106117393A
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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
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H5/00—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium
- C07H5/04—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium to nitrogen
- C07H5/06—Aminosugars
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Abstract
The invention discloses a kind of method preparing oligochitosan, comprise the steps: 1) preparation of Pupa bombycis；2) Pupa bombycis pretreatment；3) defat fat；4) deproteinization；5) de-inorganic salt；6) depigmentation；7) deacetylated；8) enzymolysis.The present invention uses exarate pupa f_1 hybrid eggs of silkworm to raise, can obtain a large amount of high-quality, free of contamination oligochitosan prepares Pupa bombycis raw material, oligochitosan extraction process is through optimizing, reduce the use ratio of the chemical reagent such as acid, alkali to greatest extent, use zymohydrolysis extracting method simultaneously, effectively controlling Environment pollution and energy resource consumption, the present invention is significantly higher than traditional method to the extraction efficiency of oligochitosan.
Description
Technical field
The present invention relates to chemical products production technical field, more particularly relate to a kind of method preparing oligochitosan.
Background technology
Oligochitosan is (also known as oligomerization aminoglucose, chitin oligosaccharide, ChitosanOligosaccharide, Chito-
Oligosaccharide, Oligochitosan) obtained by the product degradation of chitosan that chitin (chitin) is deacetylated, be
The oligosaccharide being formed by connecting by β-1-4 glycosidic bond by 2～10 aminoglucoses, is also unique a large amount of alkali existed in natural sugar
Property aminopolysaccharide, good water solubility, easily absorbed by animal body, there is safety non-toxic, antibacterial, moisture absorption is ventilative, adsorptivity is strong, fibre forming property
Get well and the feature such as good chemical reactivity and biological activity, be to produce food, health product and biomedical material
Desirable feedstock.
Although chitin is a kind of excellent biological raw material, but it is combined closely as with protein and some inorganic matter
Biological structure composition, extraction is not easy to.It is used for producing chitin, chitosan (chitin deacetylase in global range at present
Product after base, can be greatly increased dissolubility) and the prevailing technology of derivant be chemical method, its preparation process uses a large amount of
Soda acid and harmful chemical reagent, produce substantial amounts of waste water waste material, serious environment pollution particularly water body environment；Frequent,
High-temperature process brings huge energy resource consumption for a long time；Washing, purifying program and diluted acid, the recovery of diluted alkaline frequently
Intractability causes again the cleaning fresh water of preciousness to be consumed waste in a large number；All this kind, sustainable for this " green material "
The commercial application of Development and Production and a deep step has coverd with heavy shade.
Producing the most frequently used raw material of chitin at present is the shrimp of industrial waste, Carapax Eriocheir sinensis, has a fixed limit on raw material uses
System.In recent years, the fact that in Insecta animal cot containing abundant chitin, cause the very big concern of people.Research confirms, family
In Pupa bombycis skin, contained chitin is up to 33-44%, and is practically free of calcareous inorganic matter of Denging in this type of insecticide cot, beneficially first
The separation and Extraction of shell element, is rather preferable chitin raw materials for production.But, recent practice have shown that, with chemical method from Pupa bombycis
Producing chitin to have difficulties, mainly show as fat in cot too many, pigment is too many, deacetylation difficulty, thus produces
Chitin product quality the highest.Additionally, due to extraction process adds chemical degreasing step, enhance decolouring with deacetylated
The physical/chemical strength disposal of base, creates bigger waste liquid, waste entrainment and more energy consumption, therefore, how to improve work
Skill, enables to utilize Pupa bombycis to develop high-quality chitin and Related product, can effectively control again energy consumption and environment simultaneously
Pollute, be the huge challenge that chitin is researched and developed and producers face.
Summary of the invention
It is an object of the invention to provide a kind of method preparing oligochitosan, extract oligochitosan with Pupa bombycis for raw material, carry
Taking efficiency high, oligochitosan purity is high.
The present invention provides a kind of method preparing oligochitosan, comprises the steps:
1) preparation of Pupa bombycis: after Silkworm incubation, normally raise, after being fond of matured silkworm, utilizes plastic folding bunch to be placed on small straw bundles to spin cocoons, is placed on small straw bundles to spin cocoons
7th day starts, and collects Pupa bombycis from folding bunch, stand-by；
2) Pupa bombycis pretreatment: dry the Pupa bombycis collected to constant weight, then pulverize, crosses 100 mesh sieves, stand-by；
3) defat fat: the Pupa bombycis powder after sieving is taked soxhlet extraction method carry out fat disengaging, using petroleum ether as
Extractant, with cable type extractor according as extraction element, in liquid-solid ratio 3: 1, under conditions of temperature 50 C, reflux, extract, 1.5h,
To white, odorless, the pupa albumen of removing fat, after drying, obtain Silkworm pupa protein；
4) deproteinization: joined by Silkworm pupa protein in NaOH solution, reacts 1h at 85 DEG C, solution is then collected by filtration
Bottom precipitation solid, dries, obtains the dried silkworm chrysalis meal after deproteinization；
5) de-inorganic salt: joined by the dried silkworm chrysalis meal after deproteinization in HCl solution and carry out immersion treatment, removes inorganic salt and becomes
Point, solution bottom precipitation solid is collected by filtration, dries, obtain the dried silkworm chrysalis meal after de-inorganic salt；
6) depigmentation: use H2O2Aqueous solution carries out the depigmentation, and the dried silkworm chrysalis meal after above-mentioned de-inorganic salt is placed in H2O2Water-soluble
Liquid processes 15min, temperature 70-80 DEG C, filters after having processed, collect solution bottom precipitation solid, dry, stand-by；
7) deacetylated: to take step 6) dried silkworm chrysalis meal that obtains joins in the NaOH solution of mass concentration 45%, at 100 DEG C
Reason 1h, is then collected by filtration solution bottom precipitation solid, after drying, stand-by；
8) enzymolysis: by step 7) deacetylated after the dried silkworm chrysalis meal that obtains join enzymolysis solution carry out enzymolysis, wherein, enzymolysis solution
PH=4.5-5.0, enzymolysis solution and the liquid-solid ratio 5: 1 of dried silkworm chrysalis meal added, hydrolysis temperature 50-55 DEG C, enzymolysis time 2h, used
Enzymolysis solution cellulase concentration is 2.0g/100mL, and lipase concentration is 1.5g/100mL, and enzymolysis filters after completing, and receives
Collection liquid bottom precipitation solid, after drying, obtains oligochitosan finished product.
Preferably, in step 1) in, silkworm egg used is exarate pupa silkworm and mountain valley with clumps of trees and bamboo pine kind silkworm hybrid generation silkworm egg.
Preferably, dry temperature and be 80 DEG C.
Preferably, in step 4) in, Silkworm pupa protein is 1: 8 with the solid-to-liquid ratio of NaOH solution, NaOH solution quality used
Concentration is 7%.
Preferably, in step 5) in, the dried silkworm chrysalis meal after deproteinization is 1: 25 with the solid-to-liquid ratio of HCl solution, HCl solution used
Mass concentration is 8%, and the temperature of immersion treatment is 55 DEG C, soak time 2h.
Preferably, in step 6) in, H2O2Aqueous solution mass concentration is 20%.
Relative to prior art, there is advantages that
The invention provides a kind of method preparing oligochitosan, use exarate pupa f_1 hybrid eggs of silkworm to raise, it is possible to obtain
A large amount of high-qualitys, free of contamination oligochitosan prepare Pupa bombycis raw material, oligochitosan extraction process through optimizing, reduce to greatest extent acid,
The use ratio of the chemical reagent such as alkali, uses zymohydrolysis extracting method simultaneously, effectively controls Environment pollution and energy resource consumption, Jing Guoben
The oligochitosan that method is extracted can reach the 5% of dried silkworm chrysalis species, compares more traditional efficiency extracting oligochitosan from shrimp, Carapax Eriocheir sinensis
2%, the present invention is significantly higher than traditional method to the extraction efficiency of oligochitosan.
Detailed description of the invention
Below in conjunction with embodiment, technical scheme is clearly and completely described, it is clear that described reality
Executing example is only a part of embodiment of the present invention rather than whole embodiments.Based on the embodiment in the present invention, this area is general
The every other embodiment that logical technical staff is obtained under not making creative work premise, broadly falls into present invention protection
Scope.
Embodiment 1
A kind of method preparing oligochitosan that the embodiment of the present invention provides, comprises the following steps:
1) preparation of Pupa bombycis: after Silkworm incubation, normally raise, after being fond of matured silkworm, utilizes plastic folding bunch to be placed on small straw bundles to spin cocoons, is placed on small straw bundles to spin cocoons
7th day starts, and collects Pupa bombycis from folding bunch, stand-by；In step 1) in silkworm egg used be that exarate pupa silkworm is miscellaneous with mountain valley with clumps of trees and bamboo pine kind silkworm
Hand over generation silkworm egg.This step is obtained in that a large amount of high-quality, free of contamination oligochitosan extract raw material, and the extraction for follow-up oligochitosan is made
Prepare for raw material；
2) Pupa bombycis pretreatment: dry the Pupa bombycis 80 DEG C collected to constant weight, then pulverize, crosses 100 mesh sieves, stand-by；To receipts
The pretreatment that the Pupa bombycis of collection is carried out, method is simple, keeps material effective component to run off, can be that follow-up high efficiency extraction establishes base
Plinth；
3) defat fat: weigh the Pupa bombycis powder 100g after sieving, takes soxhlet extraction method to carry out the disengaging of fat, with oil
Ether is as extractant, with cable type extractor according as extraction element, in liquid-solid ratio 3: 1, under conditions of temperature 50 C, reflux, extract,
1.5h, obtains white, odorless, the pupa albumen of removing fat, obtains Silkworm pupa protein after 80 DEG C of drying；This step carries out defat
Fat, de-releasing effect can reach 99%, and petroleum ether reusable edible；
4) deproteinization: joined by Silkworm pupa protein in NaOH solution, reacts 1h at 85 DEG C, solution is then collected by filtration
Bottom precipitation solid, 80 DEG C of drying, obtain the dried silkworm chrysalis meal after deproteinization；Silkworm pupa protein is 1: 8 with the solid-to-liquid ratio of NaOH solution,
NaOH solution mass concentration used is 7%；The method for removing protein of this step, simple to operate, deproteinization effect can reach 95% with
On；
5) de-inorganic salt: joined by the dried silkworm chrysalis meal after deproteinization in HCl solution and carry out immersion treatment, removes inorganic salt and becomes
Point, solution bottom precipitation solid, 80 DEG C drying are collected by filtration, obtain the dried silkworm chrysalis meal after de-inorganic salt；Dried silkworm chrysalis meal after deproteinization
Being 1: 25 with the solid-to-liquid ratio of HCl solution, HCl solution mass concentration used is 8%, and the temperature of immersion treatment is 55 DEG C, during immersion
Between 2h；In this step, the dried silkworm chrysalis meal after deproteinization is taken off inorganic salt, react relatively mild, energy efficient, organic acid usage amount
Few, it is possible to reduce environmental pollution；
6) depigmentation: use H2O2Aqueous solution carries out the depigmentation, and the dried silkworm chrysalis meal after above-mentioned de-inorganic salt is placed in H2O2Water-soluble
Liquid processes 15min, temperature 70 C, filters after having processed, collection solution bottom precipitation solid, 80 DEG C of drying, stand-by；Used
H2O2Aqueous solution mass concentration is 20%；Adopting the depigmentation in this way, the response time is short, and efficiency is high.
7) deacetylated: to take step 6) dried silkworm chrysalis meal that obtains joins in the NaOH solution of mass concentration 45%, at 100 DEG C
Reason 1h, is then collected by filtration solution bottom precipitation solid, after drying, stand-by；The present invention is deacetylated thoroughly, reaches 99%；
8) enzymolysis: by step 7) deacetylated after the dried silkworm chrysalis meal that obtains join enzymolysis solution carry out enzymolysis, wherein, enzymolysis solution
PH=4.5, enzymolysis solution and the liquid-solid ratio 5: 1 of dried silkworm chrysalis meal added, hydrolysis temperature 50 DEG C, enzymolysis time 2h, enzymolysis solution used
Cellulase concentration is 2.0g/100mL, and lipase concentration is 1.5g/100mL, and enzymolysis filters after completing, and collects liquid
Bottom precipitation solid, after 80 DEG C dry, obtains oligochitosan finished product.Adopt and in this way the dried silkworm chrysalis meal obtained after deacetylated is carried out
Enzymolysis, reaction regulation gentleness, reaction efficiency is high.
Embodiment 2
The method that the present embodiment prepares oligochitosan is same as in Example 1, the difference is that only: in step 6) in, H2O2Water
The temperature that solution processes is 75 DEG C；In step 8) in, enzymolysis solution pH=4.8 used, hydrolysis temperature is 52 DEG C.
Embodiment 3
The method that the present embodiment prepares oligochitosan is same as in Example 1, and difference is: in step 6) in, H2O2Water-soluble
The temperature that liquid processes is 80；In step 8) in, enzymolysis solution pH=5.0 used, hydrolysis temperature is 55 DEG C.
The method using embodiment of the present invention 1-3, weighs the dried silkworm chrysalis meal 100g obtained after Pupa bombycis pretreatment respectively, carries out
Three groups of tests, after experiment, three groups of tests obtain finished product oligochitosan content and are respectively 4.85g, 4.72g, 4.91g, utilize light splitting light
In degree method detection finished product oligochitosan, effective component content is respectively 4.75%, 4.64%, 4.74%, and impurity content is below 2%.
Result of the test is as shown in table 1.By test, illustrate to utilize the preparation method extraction oligochitosan of the present invention, it is thus achieved that oligochitosan pure
The impurity such as degree is high, albumen are few, and product shows white, tasteless powder, defat fat, decolorizing effect are preferable.
Testing result after the extraction of table 1 oligochitosan
Shown in sum up, a kind of method preparing oligochitosan that the embodiment of the present invention provides, use exarate pupa silkworm first cross
Planting and raise, it is possible to obtain a large amount of high-qualitys, free of contamination oligochitosan prepare Pupa bombycis raw material, oligochitosan extraction process is passed through and is optimized,
The use ratio reducing the chemical reagent such as acid, alkali of limits, uses zymohydrolysis extracting method simultaneously, effectively control Environment pollution with
Energy resource consumption, the oligochitosan extracted through this method can reach the 5% of dried silkworm chrysalis species, compares more traditional from shrimp, Carapax Eriocheir sinensis
Extracting the efficiency 2% of oligochitosan, the present invention is significantly higher than traditional method to the extraction efficiency of oligochitosan.
Although preferred embodiments of the present invention have been described, but those skilled in the art once know basic creation
Property concept, then can make other change and amendment to these embodiments.So, claims are intended to be construed to include excellent
Select embodiment and fall into all changes and the amendment of the scope of the invention.
Obviously, those skilled in the art can carry out various change and the modification essence without deviating from the present invention to the present invention
God and scope.So, if these amendments of the present invention and modification belong to the scope of the claims in the present invention and equivalent technologies thereof
Within, then the present invention is also intended to comprise these change and modification.
Claims (6)
1. the method preparing oligochitosan, it is characterised in that comprise the steps:
1) preparation of Pupa bombycis: after Silkworm incubation, normally raise, after being fond of matured silkworm, utilizes plastic folding bunch to be placed on small straw bundles to spin cocoons, and is placed on small straw bundles to spin cocoons the 7th
It starts, and collects Pupa bombycis from folding bunch, stand-by；
2) Pupa bombycis pretreatment: dry the Pupa bombycis collected to constant weight, then pulverize, crosses 100 mesh sieves, stand-by；
3) defat fat: the Pupa bombycis powder after sieving is taked soxhlet extraction method carry out the disengaging of fat, using petroleum ether as extraction
Solvent, with cable type extractor according as extraction element, in liquid-solid ratio 3: 1, under conditions of temperature 50 C, reflux, extract, 1.5h, obtains white
Color, odorless, the pupa albumen of removing fat, obtain Silkworm pupa protein after drying；
4) deproteinization: joined by Silkworm pupa protein in NaOH solution, reacts 1h at 85 DEG C, is then collected by filtration bottom solution
Precipitation solid, dries, obtains the dried silkworm chrysalis meal after deproteinization；
5) de-inorganic salt: the dried silkworm chrysalis meal after deproteinization is joined in HCl solution and carry out immersion treatment, remove inorganic salts ingredients,
Solution bottom precipitation solid is collected by filtration, dries, obtain the dried silkworm chrysalis meal after de-inorganic salt；
6) depigmentation: use H2O2Aqueous solution carries out the depigmentation, and the dried silkworm chrysalis meal after above-mentioned de-inorganic salt is placed in H2O2In aqueous solution
Process 15min, temperature 70-80 DEG C, filter after having processed, collect solution bottom precipitation solid, dry, stand-by；
7) deacetylated: to take step 6) dried silkworm chrysalis meal that obtains joins in the NaOH solution of mass concentration 45%, in 100 DEG C of process
1h, is then collected by filtration solution bottom precipitation solid, after drying, stand-by；
8) enzymolysis: by step 7) deacetylated after the dried silkworm chrysalis meal that obtains join enzymolysis solution carry out enzymolysis, wherein, enzymolysis solution pH=
4.5-5.0, enzymolysis solution and the liquid-solid ratio 5: 1 of dried silkworm chrysalis meal added, hydrolysis temperature 50-55 DEG C, enzymolysis time 2h, enzymolysis used
Liquid cellulase concentration is 2.0g/100mL, and lipase concentration is 1.5g/100mL, and enzymolysis filters after completing, and collects liquid
Body bottom precipitation solid, after drying, obtains oligochitosan finished product.
2. the method for claim 1, it is characterised in that in step 1) in, silkworm egg used is exarate pupa silkworm and mountain valley with clumps of trees and bamboo pine product
Plant silkworm hybrid generation silkworm egg.
3. the method for claim 1, it is characterised in that dry temperature and be 80 DEG C.
4. the method for claim 1, it is characterised in that in step 4) in, Silkworm pupa protein and the solid-liquid of NaOH solution
Ratio is 1: 8, and NaOH solution mass concentration used is 7%.
5. the method for claim 1, it is characterised in that in step 5) in, the dried silkworm chrysalis meal after deproteinization and HCl solution
Solid-to-liquid ratio is 1: 25, and HCl solution mass concentration used is 8%, and the temperature of immersion treatment is 55 DEG C, soak time 2h.
6. the method for claim 1, it is characterised in that in step 6) in, H2O2Aqueous solution mass concentration is 20%.
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Cited By (1)
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| CN107411098A (en) * | 2017-04-22 | 2017-12-01 | 安徽省农业科学院蚕桑研究所 | A kind of health products preparation method containing natural silkworm pupa protein polypeptide |
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| CN107411098A (en) * | 2017-04-22 | 2017-12-01 | 安徽省农业科学院蚕桑研究所 | A kind of health products preparation method containing natural silkworm pupa protein polypeptide |
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Application publication date: 20161116 |