CN106755196A - One kind improves the water miscible method of beta glucan - Google Patents
One kind improves the water miscible method of beta glucan Download PDFInfo
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- CN106755196A CN106755196A CN201611186178.5A CN201611186178A CN106755196A CN 106755196 A CN106755196 A CN 106755196A CN 201611186178 A CN201611186178 A CN 201611186178A CN 106755196 A CN106755196 A CN 106755196A
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- beta glucan
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- glucan
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- 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
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- 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
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
- C12P19/08—Dextran
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Abstract
The application belongs to polysaccharide dissolving improving environment field, and in particular to one kind improves the water miscible method of beta glucan.A kind of raising water miscible method of beta glucan provided by the present invention, while being degraded to beta glucan using ultrasonic technology and enzymatic isolation method, has played the synergy of ultrasonic wave and enzymatic isolation method, substantially increases the water solubility of beta glucan.The present invention has the advantages that operating procedure is simple, feasibility by force, without using organic solvent, environmental protection, low production cost, for the production of highly-water-soluble yeast beta glucan provides feasible method, and the inventive method on the bioactivity of beta glucan without influence, in can be widely applied to food and biomedicine field.
Description
Technical field
The invention belongs to polysaccharide dissolving improving environment field, and in particular to one kind improves the water miscible method of beta glucan,
More particularly to it is a kind of using the ultrasound-assisted enzymolysis method raising water miscible method of yeast beta-dextran.
Background technology
Beta glucan is a kind of active polysaccharide being widely present in microorganism, mushroom and plant, and one of important sources are
Saccharomyces cerevisiae.Though China possesses abundant saccharomyces cerevisiae resource, but develop the waste yeast of generation rapidly for brewing industry
Recycle, be typically limited to only and it is simply processed, and be mainly used in the production of field of fodder or yeast preparation, lead
Largely discarded saccharomyces cerevisiae is caused with discharge of wastewater, to cause the waste of resource and the severe contamination of environment.
With the fast development and the raising of people's level of consumption of functional food industry, the beta glucan of Saccharomyces cerevisiae
Because of active effect of its uniqueness, increasingly cause the concern of people, can be used as a kind of active functional component of green natural.In recent years
Come, the American-European nutraceutical for waiting some developed countries that various infants, old age and pregnant woman are developed using beta glucan, and with
The positive cumulative year after year of the output value of the dietary fiber food based on beta glucan, therefore before beta glucan functional product has fabulous market
Scape.However, the saccharomyces cerevisiae beta glucan for obtaining directly is extracted because it is in neutrality by existing conventional Polyose extraction technology,
Neutral, and intramolecular polyhydroxy group interaction, form densification triple helix structure, in aggregated particle shape,
Significantly limit beta glucan.Therefore, chemical modification or biodegradation are carried out to it, improves dissolubility, be to develop ferment
The effective way of female glucan.
At present, mainly by chemistry, biological or physical modification etc., method is water miscible to beta glucan changes for domestic and foreign scholars
It is kind to be studied.Wherein, chemical modification increases solubility, such as phosphoric acid by connecting hydrophilic radical on yeast dextran
Change, carboxy methylation, Sulfation modification, this kind of reaction is carried out in organic solvent system, seriously polluted, there is potential safety hazard.
Biological modified method complex operation, it is costly.It is a process for randomness that physics decorations are modified, and it reduces relative molecular mass
Be limited in scope.Therefore, those skilled in the art need a kind of easy and effective, feasibility method high badly for improving beta glucan
It is water-soluble.
The content of the invention
In view of this, the water miscible method of beta glucan is improved it is an object of the invention to provide one kind, it is existing for solving
There is the big technical problem of complex operation in technology, organic solvent consumption, the water solubility of beta glucan can be increased substantially and be
The production of highly-water-soluble yeast beta-dextran provides feasible method.
Concrete technical scheme of the invention is as follows：
The water miscible method of beta glucan is improved the invention provides one kind, including：Beta glucan solution and β-Portugal are gathered
Carbohydrase mixes, and is digested under ul-trasonic irradiation.
Preferably, 1,4 beta-glucanase described in 2~4U is added in every milliliter of beta glucan solution.
Preferably, the temperature of the enzymolysis is 40~50 DEG C；The time of the enzymolysis is 1~2h.
Preferably, the power of the ultrasonic wave is 100~200W.
Preferably, the beta glucan solution is prepared as：Beta glucan is dissolved in NaAc_HAc buffer solution
In.
Preferably, the mixed proportion of the beta glucan and the NaAc_HAc buffer solution is 1 μ g：(10-20)
mL。
Preferably, the concentration of the NaAc_HAc buffer solution is 0.1~1mol/L.
It is furthermore preferred that the dissolving includes：Water-bath preheats 15~20min at 35~38 DEG C.
Beta glucan is the low active polysaccharide of the active strong, toxic and side effect of a class.In to beta glucan modifying process, play
The ultrasonic wave of physical action weakens the glycosidic bond of macromolecular main chain by the mechanical function of particle vibration, promotes beta glucan solution spiral shell
Rotation, meanwhile, acted on by digestion with reference to 1,4 beta-glucanase and be broken the glycosidic bond of beta glucan so that macromolecular glucan is hydrolyzed
It is smaller oligosaccharides or reduced sugar so as to increase dissolubility of the beta glucan in water.Meanwhile, the various lifes of molecular structure and polysaccharide
Thing activity has close relationship, and in modifying process, biologic enzymolysis method mild condition reduces beta glucan by digestion
The degree of polymerization does not change its senior molecular structure to the physical action of beta glucan without changing its natural structure, ultrasonic wave, therefore
Cause that beta glucan increases its dissolubility on the basis of original biology natural structure is kept using ultrasound-assisted enzymolysis method, because
Bioactivity influence of this inventive method on beta glucan is relatively low or even without influence.
The water miscible method of beta glucan is improved the invention provides one kind, while using ultrasonic technology and enzymatic isolation method pair
Beta glucan is degraded, and has played the synergy of ultrasonic wave and enzymatic isolation method, substantially increases the water solubility of beta glucan.This
Invention has the advantages that operating procedure is simple, feasibility strong, without using organic solvent, environmental protection, low production cost, be height
The production of water-soluble yeast beta-dextran provides feasible method, and the inventive method to the bioactivity of beta glucan without shadow
Ring, in can be widely applied to food and biomedicine field.
Specific embodiment
Technical scheme is clearly and completely described below in conjunction with the preferred embodiment of the present invention, it is clear that
Described embodiment is a part of embodiment of the invention, rather than whole embodiments.Those skilled in the art should manage
Solution, modifies to specific embodiment of the invention or some technical characteristics is replaced on an equal basis, without deviating from the present invention
The spirit of technical scheme, all should cover in the scope of protection of the invention.
Beta glucan employed in following examples is the beta glucan of Saccharomyces cerevisiae, biological purchased from Shanghai source leaf
Science and Technology Ltd.；1,4 beta-glucanase is purchased from Shanghai Yuan Ye bio tech ltd.
Embodiment 1
The first step, 1 μ g are pressed by beta glucan：The mixed proportion of 10mL is invested in NaAc_HAc buffer solution
(0.1mol/L, pH5.0), 35 DEG C of water-bath preheating 15min are made beta glucan solution；
Second step, it is the 1,4 beta-glucanase of 2U/mL to add [E/S], wherein, [E/S] represents that 1,4 beta-glucanase is poly- in β-Portugal
Addition concentration in sugar juice；
3rd step, is placed in supersonic generator (model SB-600DTY), is the subsidiary conditions of 100W in ultrasonic power
Under, 40 DEG C of constant temperature digest 1h；
4th step, boiling water bath goes out enzyme activity 5min, with the centrifugation 5min of 4000r/min, the supernatant for obtaining as β-
Glucan aqueous solution.
Using polysaccharide and reduced sugar in phend-sulphuric acid and the Somogyi-Neslon sequentially determining beta glucan aqueous solution
Content, calculates soluble polysaccharide yield, and its computing formula is as follows：
Soluble polysaccharide=total Soluble Sugar (GT)-reduced sugar (GR)；
Soluble polysaccharide yield (%)=100 × soluble polysaccharide (g)/glucan dry weight (g) × 100%.
The present embodiment improves the water-soluble of beta glucan using the modified saccharomyces cerevisiae beta glucan of ultrasound-assisted enzymolysis method
Property, it has been respectively increased 15% and 10% when its soluble polysaccharide yield is than being used alone ultrasonic degradation and 1,4 beta-glucanase enzymolysis.
Embodiment 2
The first step, 1 μ g are pressed by beta glucan：The mixed proportion of 15mL is invested in NaAc_HAc buffer solution
(0.5mol/L, pH5.0), 35 DEG C of water-bath preheating 18min are made beta glucan solution；
Second step, it is the 1,4 beta-glucanase of 3U/mL to add [E/S]；
3rd step, is placed in supersonic generator (model SB-600DTY), is the subsidiary conditions of 150W in ultrasonic power
Under, 45 DEG C of constant temperature digest 1.5h；
4th step, boiling water bath goes out enzyme activity 8min, with the centrifugation 8min of 4500r/min, the supernatant for obtaining as β-
Glucan aqueous solution.
Using polysaccharide and reduced sugar in phend-sulphuric acid and the Somogyi-Neslon sequentially determining beta glucan aqueous solution
Content, calculates soluble polysaccharide yield, and its computing formula is as follows：
Soluble polysaccharide=total Soluble Sugar (GT)-reduced sugar (GR)；
Soluble polysaccharide yield (%)=100 × soluble polysaccharide (g)/glucan dry weight (g) × 100%.
The present embodiment improves the water-soluble of beta glucan using the modified saccharomyces cerevisiae beta glucan of ultrasound-assisted enzymolysis method
Property, it has been respectively increased 18% and 12% when its soluble polysaccharide yield is than being used alone ultrasonic degradation and 1,4 beta-glucanase enzymolysis.
Embodiment 3
The first step, 1 μ g are pressed by beta glucan：The mixed proportion of 20mL is invested in NaAc_HAc buffer solution (1mol/
L, pH5.0), 38 DEG C of water-bath preheating 20min are made beta glucan solution；
Second step, it is the 1,4 beta-glucanase of 4U/mL to add [E/S]；
3rd step, is placed in supersonic generator (model SB-600DTY), is the subsidiary conditions of 200W in ultrasonic power
Under, 50 DEG C of constant temperature digest 2h；
4th step, boiling water bath goes out enzyme activity 10min, with the centrifugation 8min of 5000r/min, the supernatant for obtaining as β-
Glucan aqueous solution.
Using polysaccharide and reduced sugar in phend-sulphuric acid and the Somogyi-Neslon sequentially determining beta glucan aqueous solution
Content, calculates soluble polysaccharide yield, and its computing formula is as follows：
Soluble polysaccharide=total Soluble Sugar (GT)-reduced sugar (GR)；
Soluble polysaccharide yield (%)=100 × soluble polysaccharide (g)/glucan dry weight (g) × 100%.
The present embodiment improves the water-soluble of beta glucan using the modified saccharomyces cerevisiae beta glucan of ultrasound-assisted enzymolysis method
Property, it has been respectively increased 20% and 15% when its soluble polysaccharide yield is than being used alone ultrasonic degradation and 1,4 beta-glucanase enzymolysis.
In sum, a kind of raising water miscible method of beta glucan provided by the present invention, using ultrasound wave auxiliary enzyme
Solution is degraded to beta glucan, and its catabolite soluble polysaccharide yield is higher during one of which biodegrading process than being used alone
10~20%, ultrasonic wave and enzymatic isolation method synergy in the inventive method are illustrated, the water solubility of beta glucan is substantially increased, reach
1+1 is arrived>2 beneficial effect.The inventive method has that operating procedure is simple, feasibility strong, without using organic solvent, green
Environmental protection and the advantage of low production cost, the beta glucan that the water-soluble more single method of modifying of beta glucan of acquisition is obtained are water-soluble
Height, and its bioactivity keeps, in can be widely applied to food and biological medicine industry.
Claims (7)
1. one kind improves the water miscible method of beta glucan, including：Beta glucan solution and 1,4 beta-glucanase are mixed, in ultrasound
Digested under ripple effect.
2. method according to claim 1, it is characterised in that 2~4U institutes are added in every milliliter of beta glucan solution
State 1,4 beta-glucanase.
3. method according to claim 1, it is characterised in that the temperature of the enzymolysis is 40~50 DEG C；The enzymolysis
Time is 1~2h.
4. method according to claim 1, it is characterised in that the power of the ultrasonic wave is 100~200W.
5. method according to claim 1, it is characterised in that the beta glucan solution is prepared as：By beta glucan
It is dissolved in NaAc_HAc buffer solution.
6. method according to claim 5, it is characterised in that the beta glucan and Acetic acid-sodium acetate buffering are molten
The mixed proportion of liquid is 1 μ g：(10-20)mL.
7. method according to claim 5, it is characterised in that the concentration of the NaAc_HAc buffer solution is 0.1~
1mol/L。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201611186178.5A CN106755196A (en) | 2016-12-20 | 2016-12-20 | One kind improves the water miscible method of beta glucan |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611186178.5A CN106755196A (en) | 2016-12-20 | 2016-12-20 | One kind improves the water miscible method of beta glucan |
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| CN201611186178.5A Pending CN106755196A (en) | 2016-12-20 | 2016-12-20 | One kind improves the water miscible method of beta glucan |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112301073A (en) * | 2020-10-19 | 2021-02-02 | 上海应用技术大学 | Method for solubilizing and modifying yeast beta-glucan by ultrasonic-enzyme method |
| CN114732127A (en) * | 2020-12-23 | 2022-07-12 | 安琪酵母(柳州)有限公司 | Preparation method and application of yeast extract rich in soluble glucan |
| CN117016797A (en) * | 2023-08-14 | 2023-11-10 | 四川合泰新光生物科技有限公司 | Method for improving dissolution rate of glucan |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112301073A (en) * | 2020-10-19 | 2021-02-02 | 上海应用技术大学 | Method for solubilizing and modifying yeast beta-glucan by ultrasonic-enzyme method |
| CN114732127A (en) * | 2020-12-23 | 2022-07-12 | 安琪酵母(柳州)有限公司 | Preparation method and application of yeast extract rich in soluble glucan |
| CN117016797A (en) * | 2023-08-14 | 2023-11-10 | 四川合泰新光生物科技有限公司 | Method for improving dissolution rate of glucan |
| CN117016797B (en) * | 2023-08-14 | 2024-06-04 | 四川合泰新光生物科技有限公司 | Method for improving dissolution rate of glucan |
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