CN101353383B - Water-soluble yeast beta-dextran and preparation thereof - Google Patents
Water-soluble yeast beta-dextran and preparation thereof Download PDFInfo
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Abstract
The invention relates to a water-soluble barm Beta-dextran and a preparation method thereof. In the water-soluble barm Beta-dextran chain, Beta-1, 3-dextran is taken as a main chain, Beta-1, 6-dextran is taken as a branched chain, the molecular weight is 0.08 to 0.2 million Daltun. The preparation method of the water-soluble barm Beta-dextran includes the following steps of: water extraction, alkali extraction and impurity processing. The preparation technique of the invention is simple; the technique parameters are easy to be operated and controlled; both the yield and the purity of the product water-soluble barm Beta-dextran are higher. The obtained water-soluble barm Beta-dextran has remarkable immune activation effect and especially can improve the phagocytic function and the antibodyformation capacity of a macrophage to a large extent, thereby improving the body immunity and resistance. The water-soluble barm Beta-dextran can be widely applied in the fields such as food, dairy food and drinks, etc.
Description
Technical field:
The present invention relates to a kind ofly prepare the method for zymosan, specially refer to a kind of preparation method of water-soluble yeast beta-dextran by yeast.
Background technology:
Yeast is a kind of very important industrial microorganism, and most of yeast carry out cheapness as roughage to be handled, and does not do further deep processing, can not form the real comprehensive utilization to by product.Part enterprise with its direct discharging, has caused the great wasting of resources and environmental pollution especially.
β-1, the 3-dextran is prevalent in multiple fungi, plant and the alginic cell wall, is a kind of bioactive immune polysaccharide that is rich in, and having confirmed all has better therapeutic effect to difficult disease such as tumour, hepatitis, diabetes, cardiovascular, reducing blood-fat.β-1, the 3-dextran still is a kind of good biological response modifiers (BRMs), and it can improve the immunological competence of body by stimulating immune system, and BRMs is in complementary airframe systems, to keeping homeostasis is very crucial, and it helps organism adaptation environment and psychological pressure.
Except that containing amounts of protein, also contain mannosans, Mannoproteins, β-1 in the yeast cell, compositions such as 3-dextran, other mixed polysaccharide and chitin.The zymosan product that has been employed at present mainly contains mannosans, mannooligo saccharide, the Mannoproteins of seminose series, contains the zymosan of multiple polysaccharide component, non-water-soluble yeast cell wall beta-1,3-D-dextran etc.Water-soluble yeast beta-1 is not arranged as yet, the production of 3-dextran and application.
Utilize yeast cells wall to extract β-1, the method of 3-dextran is more, mainly contain acid system extraction, alkaline process extraction, soda acid one method, organic solvent extraction, self-dissolving-enzyme-alkaline process, ultrasonic extraction, enzyme process etc., the β-1 of common gained, the 3-dextran is water-insoluble yeast glucan, molecular weight is huge, the component that is difficult to absorb for human body.In Chinese patent (CN:1583802A), disclosed a kind of β-1, the preparation method of 3-dextran, the yeast β-1 that this method adopts alkaline extraction, acid extraction, organic solvent lixiviate to obtain, the 3-dextran, be water-insoluble yeast glucan, to this β-1, the 3-dextran is degraded by formic acid again, obtain the water-soluble yeast beta-1 of different molecular weight, the 3-dextran; In another Chinese patent (CN:101012468), disclosed a kind of β-1, the yeast β-1 that the preparation method of 3-dextran, this method adopt high temperature extraction, enzyme processing, organic solvent lixiviate to obtain, the 3-dextran also is a kind of water-insoluble yeast glucan.This type of yeast β-1, the content of 3-dextran is lower, generally about 70%, contains multiple abiotic active impurity.Because non-water-soluble yeast β-1,3-dextran molecule amount height can not be dissolved in water, and not have beta-glycosidase in the human body, is difficult to directly absorb, and can only pass through the enteric microorganism degraded and absorbed, greatly reduces yeast β-1, the bioavailability of 3-dextran.
At present, resulting yeast β-1 in the various reports, the 3-dextran mostly is non-water-soluble yeast dextran, and ISG is owing to be insoluble in water, and its range of application is subjected to very big restriction.The water-soluble yeast β-1 of part, the 3-dextran also is to obtain by water-insoluble yeast glucan degraded or modification.Through the few dextran or the dextran through modifying of degraded, though solvability improves, preparation cost improves greatly, has limited the extensive application in some field equally.
Summary of the invention:
At many deficiencies that existing yeast beta-dextran exists, the invention provides a kind of is water-soluble yeast beta-dextran of feedstock production and preparation method thereof with the complete thalline of yeast, and the molecular weight of this dextran is 8-20 ten thousand Dalton, can be water-soluble fully.Preparation method's technology of the water-soluble yeast beta-dextran that is provided is simple, productive rate is enhanced than prior art, and yeast has been reached maximum utilization.
Yeast beta-dextran of the present invention is that raw material makes with complete yeast, is β-1, the 3-D-dextran, with β-1, the 3-dextran is a main chain, β-1, and the 6-dextran is a side chain, its weight average molecular weight range is 8-20 ten thousand Dalton, generally can reach more than 100,000 Dalton,, highly purified after, its weight-average molecular weight is 12-15 ten thousand Dalton, can be water-soluble fully.
The preparation of beta-glucan of the present invention realizes by processes such as hot-water cure, alkaline extraction, acid neutralization, alcohol precipitation, column chromatographies.Residue to behind the preparation water-soluble yeast beta-dextran obtains high-load water-insoluble yeast beta-dextran by enzymolysis again, and concrete steps are as follows:
1. hot-water cure: in yeast powder or fresh yeast, add pure water, making thalline weight is the preferred 5-10% (w/v) of 2-20% (w/v) of gross weight, wherein when using fresh yeast, the weight of fresh yeast is converted to dry yeast weight to be calculated and gets final product, be warmed up to 50-121 ℃, stirring reaction 0.5-2h, 3000-12000r/min is centrifugal afterwards, collecting precipitation thing I; Clear liquid is evaporated to the 1/3-1/15 of original volume, adds ethanol, and this alcohol concn is 80-100% (v/v), makes the determining alcohol in the clear liquid reach 60-80% (v/v) alcohol precipitation, makes precipitation, and this throw out is a mannosans;
2. alkaline purification: add alkaline solution in throw out I, the alkaline solution volume of adding is preferably 10 times for 2-30 times of precipitation I volume, and alkaline concentration is 3-10% (w/v), and its concentration is preferably 4%-10%, optimal selection 4%; Be warmed up to 60-85 ℃ afterwards, stirring reaction 0.5-2h, 3000-12000r/min is centrifugal, collects alkali and carries throw out III; Clear liquid neutralizes with acid, be neutralized to about pH5.0-8.0, preferably be neutralized to pH6.5-7.5,3000-12000r/min is centrifugal afterwards, and clear liquid is concentrated into the 1/3-1/15 of original volume, preferably be concentrated into about 1/5 of original volume, generally remain on 1/4-1/6, add ethanol, this alcohol concn is 80-100% (v/v), make determining alcohol reach 60-85%, obtain water-soluble yeast beta-dextran precipitation II;
After this throw out II separation redissolution, separate through column chromatography again, desalination, drying obtains the high purity water-soluble yeast beta-dextran; It is eluents commonly used such as salts solution or water that column chromatography is separated the eluent that is adopted;
3. enzyme is handled: carry the water that adds 5-20 times of volume among the throw out III to alkali, this throw out is suspended in the aqueous solution, regulate pH4.0-9.0 with alkali, add proteolytic enzyme, make the proteolytic enzyme total amount reach the 0.02-0.2% (W/V) of suspension total amount, be warmed up to 35-60 ℃, reaction times 2-24h, the 3000-12000r/min centrifugal collecting precipitate to this throw out water thorough washing, is non-water-soluble yeast beta-dextran afterwards.
After adopting above-mentioned enzymolysis process to handle, the content of water-insoluble yeast beta-dextran can reach more than 90%.
In the above-mentioned technological process, the employed alkali of described alkaline purification process is sodium hydroxide or potassium hydroxide or yellow soda ash or its mixture; In the described alkaline purification process and the time acid used be citric acid or acetate or hydrochloric acid or sulfuric acid or its mixture; The column chromatography separating medium that adopts when column chromatography is separated is DOWEX MXA-1 or Q-Sepharose FF or DEAE-Sephacel or DEAE-Sepharose FF.
Regulating the used alkali of PH in the described enzyme treating processes is sodium hydroxide or potassium hydroxide or yellow soda ash or its mixture; The proteolytic enzyme that is adopted is neutral protease or Sumizyme MP or papoid or its mixture.
The preparation method of above-mentioned beta-glucan, the optimum condition of variety of processes is:
The optimum condition of hot water extraction process is: the body weight of yeast described in the hot water extraction is the 10-15% (w/v) of pure water and yeast thalline gross weight; Temperature 80-100 ℃, the reaction times is 1-2h;
Optimum condition in the alkaline extraction process is: the alkali that is adopted is preferably sodium hydroxide or yellow soda ash or its mixture, concentration 4-6% (w/v), and the alkali lye consumption is 10-15 times of precipitation I volume; Temperature 60-75 ℃, the reaction times is 1-2h; In and the time acid adopted be citric acid or acetate or its mixture, its concentration is 30-100% (w/v);
The optimum condition of described enzyme treating processes is: enzyme is a Sumizyme MP, and concentration is 0.1-0.2% (w/v); Temperature 45-60 ℃, the reaction times is 8-16h.
The preparation method of above-mentioned beta-glucan, the most preferably condition of variety of processes is:
The most preferably condition of leaching process is: be 10% (w/v) of pure water and yeast thalline gross weight in yeast body weight described in the hot water extraction process; 80 ℃ of temperature, the reaction times is 1h;
Most preferably condition in the alkaline extraction process is: the alkali that is adopted is preferably sodium hydroxide, concentration 4% (w/v), and the alkali lye consumption is 10 times of precipitation I volume; 60 ℃ of temperature, the reaction times is 1h; In and the time acid adopted be citric acid, its concentration is 30% (w/v);
The most preferably condition of described enzyme treating processes is: enzyme is a Sumizyme MP, and concentration is 0.1% (w/v); 60 ℃ of temperature, the reaction times is 16h.
Under above-mentioned optimum condition, the yield and the beta-glucan content of water-soluble yeast dextran the highest (just the purity of the dextran yield of unit raw material and gained dextran is the highest) reaches 15% and 90% (w/w) respectively.
Adopt above-mentioned processing condition to prepare water-soluble yeast beta-dextran, have following outstanding advantage:
The employing alkaline purification can be with the more stripping of water-soluble yeast beta-dextran, the concentration of lye of employing 4% and 60 ℃ temperature of reaction, be in order to guarantee in the more stripping of water-soluble yeast beta-dextran, reduce the degraded destruction of alkaline solution to water-soluble yeast beta-dextran as far as possible, sodium hydroxide stripping than existing employing 2%, the solubility rate of beta-glucan improves a lot, but being unlikely to destroy the basic structure of beta-glucan, the increase of sodium hydroxide concentration does not simultaneously increase a lot of costs; Adopt in the acid and the time, the preferred citric acid that uses is the ternary organic acid, can react with the molar mass mark of sodium hydroxide with 1: 3, thereby significantly reduced sour consumption, the amount of the Citrate trianion that generates also reduces thereupon, reduce the content of impurity in the product, improved purity, reduced production cost; On the other hand, use citric acid can not bring offending flavour and mouthfeel to target product, simultaneously in the alkaline purification process owing to improved the alkali concn of alkaline purification, reduced the temperature and time of handling, so just reduced owing to the degree of damage of high temperature for target product, shorten the time of entire treatment simultaneously, improved the efficient of producing.
Adopting enzyme to handle to the residual II I behind the alkaline extraction can dispose foreign protein wherein, further improved the content of water-insoluble beta-glucan, guarantee the quality of purpose product better, than conventional in the market beta-glucan product, be that purity or quality aspect all are greatly improved.
The result who the water-soluble yeast beta-dextran that is obtained is carried out structure determination is as follows:
By thin layer chromatography analysis and Infrared spectroscopy as can be known, the water-soluble yeast polysaccharide that adopts aforesaid method to make is made up of single glucose; 3319-3329cm in the infrared spectrogram
-1, 2922cm
-1, 1039-1076cm
-1, 1157cm
-1, 889cm
-1, 770cm
-1Article six, key band explanation obtained product be β-1, the 3-D-dextran is characterized in that in its sugar chain that with β-1, the 3-dextran is a main chain, β-1, the 6-dextran is a side chain; Utilize the dextran typical curve of series standard molecular weight, measure many batch samples by the high performance liquid phase gel chromatography, its weight average molecular weight range is 8-20 ten thousand Dalton, and to highly purified sample determination, its weight-average molecular weight is 12-15 ten thousand Dalton.Its molecular weight has had very significant reduction than the molecular weight of ISG, the more water-insoluble yeast beta-dextran of its biological activity and bioavailability is greatly increased, simultaneously, also has this outstanding feature of good water-solubility, make yeast beta-dextran can directly apply in the liquid material, need not to use stablizer can obtain even and stable solution, enlarged the use range of yeast beta-dextran.
Adopt above-mentioned preparation method to prepare water-soluble yeast beta-dextran, also have another outstanding advantage:
When obtaining water-soluble yeast beta-dextran, protein-contg mannosans and conventional water-insoluble yeast beta-dextran have also been obtained, effectively the various effective constituents in the yeast thalline are recycled, when improving yield, farthest improved saccharomycetic utilization ratio.Handle three technological processs of carrying out continuously through hot water extraction, alkaline extraction and enzyme, obtained mannosans, water-soluble yeast beta-dextran and conventional yeast beta-dextran in succession.Wherein, hot-water cure process and mannosans that the enzyme treating processes obtains and water-insoluble polysaccharide are preparation technology's of the present invention by product.Owing to adopt after present method under the prerequisite that guarantees the water-soluble yeast beta-dextran yield, mannosans and water-insoluble beta-glucan have also been obtained simultaneously, so just changed traditional technology to having to the utilizable mode of portioned product in the saccharomycetic processing, improved added value of product, realized saccharomycetic deep processing, the direct discharging of waste products has been avoided in the comprehensive utilization that formation is real to yeast, has reduced the wasting of resources and to the pollution of environment.
Preparation technology of the present invention is simple, and the preparation production process can be carried out continuously, the control of processing parameter easy handling, and the yield of product water-soluble yeast beta-dextran, purity are all than higher.Resultant water-soluble yeast beta-dextran has significant immuno-stimulating effect, especially phagolysis, phagocytic index and the antibody-producing capacity to scavenger cell can increase substantially, and then the immunizing power and the resistibility of raising body, can be widely used in fields such as food, dairy products, beverage.
Description of drawings:
Fig. 1 prepares the process flow sheet of water-soluble yeast beta-dextran for the present invention;
Fig. 2 is the thin-layer chromatogram of the water-soluble yeast beta-dextran of the present invention's preparation;
The position of selecting of 1-8 is respectively among Fig. 2: semi-lactosi standard specimen, pectinose standard specimen, glucose standard specimen, embodiment 1 sample, embodiment 2 samples, embodiment 3 samples, seminose standard specimen, wood sugar standard specimen;
Fig. 3 is the infrared spectrogram of the embodiment of the invention 1 preparation water-soluble yeast beta-dextran.
Embodiment:
Get the 200g yeast powder, add the 2L pure water, extract 1h down with 80 ℃, the centrifugal 10min of 4000r/min, collecting precipitation I, clear liquid concentrates, alcohol precipitation, drying obtains the yeast mannosans; The sodium hydroxide solution that in precipitation I, adds 2L 4% (w/v) then, 60 ℃ of following stirring reaction 1h, the centrifugal 20min of 5000r/min collects alkali and carries precipitation III.Clear liquid is neutralized to pH 6.0 with 30% citric acid, to be cooled after room temperature, the centrifugal 10min of 5000r/min, clear liquid is concentrated into 1/5 of former clear liquid volume, add 95% ethanol and make the determining alcohol in the alcoholic solution reach 80%, alcohol precipitation obtains water-soluble yeast beta-dextran after the precipitation drying, the beta-glucan content of product is 85%, yield 15%.Alkali is carried among the precipitation III and is added 2L water, regulates pH8.0, adds Sumizyme MP 2g, 60 ℃ of enzymolysis 16h, and collecting precipitation, drying obtains non-water-soluble yeast beta-dextran.
Water-soluble yeast beta-dextran redissolves, be mixed with 2% solution, with 0.22um hollow fiber film assembly micro-filtration, with the ultrafiltration of 6KD hollow fiber film assembly, regulate about concentrated solution concentration to 1%, with Q-SepHarose-FF column chromatography purifying, use the 0.2mol/LNaCl eluant solution, obtain the pure product of water-soluble yeast beta-dextran.The beta-glucan content 98% of product, yield 2.0%.
Embodiment 2
Add the 2L pure water in the 200g yeast powder, 100 ℃ are extracted 1h, the centrifugal 10min of 4000r/min, and collecting precipitation I, clear liquid concentrates, alcohol precipitation, drying obtains the yeast mannosans; The sodium hydroxide that in precipitation I, adds 2L 10% then, 80 ℃ of stirring reaction 1h, the centrifugal 20min of 3000r/min collects alkali and carries precipitation III.Clear liquid is neutralized to pH7.0 with 10% (w/v) hydrochloric acid, to be cooled after room temperature, the centrifugal 15min of 3000r/min, the clear liquid ultrafiltration and concentration be original volume 1/3 after, add 95% ethanol and make determining alcohol reach 80%, alcohol precipitation obtains water-soluble yeast beta-dextran after the precipitation drying, the beta-glucan content 90% of product, yield 12%.Alkali is carried among the precipitation III and is added 2L water, regulates pH7.0, adds papoid 1g, 50 ℃ of enzymolysis 16h, and collecting precipitation, drying obtains non-water-soluble yeast beta-dextran 26g, content 95.1%.
Water-soluble yeast beta-dextran redissolves, be mixed with 2% solution, with 0.22um hollow fiber film assembly micro-filtration, with the ultrafiltration of 6KD hollow fiber film assembly, regulate about concentrated solution concentration to 1%, with DEAE-SepHaroseFF column chromatography purifying, water wash-out, obtain the pure product of water-soluble yeast beta-dextran.The beta-glucan content 96% of product, yield 1.1%.
Embodiment 3
Add the 1.2L pure water in the 100g yeast powder, 121 ℃ are extracted 1.5h, the centrifugal 10min of 4000r/min, and collecting precipitation I, clear liquid concentrates, alcohol precipitation, drying obtains the yeast mannosans; The sodium carbonate solution that in precipitation I, adds 2L 6% then, 60 ℃ of stirring reaction 2h, the centrifugal 8min of 12000r/min collects alkali and carries precipitation III.Clear liquid is neutralized to pH 6.0 with 10% (w/v) acetate, to be cooled after room temperature, the centrifugal 5min of 12000r/min, the clear liquid ultrafiltration and concentration be original volume 1/12 after, add 95% ethanol and make determining alcohol reach 80%, alcohol precipitation obtains water-soluble yeast beta-dextran after the precipitation drying, the beta-glucan content 85% of product, yield 10%.Alkali is carried among the precipitation III and is added 1.2L water, regulates pH8.0, adds Sumizyme MP 2g, 60 ℃ of enzymolysis 16h, and collecting precipitation, drying obtains non-water-soluble yeast beta-dextran.
Water-soluble yeast beta-dextran redissolves, and is mixed with 2% solution, with 0.22um hollow fiber film assembly micro-filtration, with the ultrafiltration of 6KD hollow fiber film assembly, regulate about concentrated solution concentration to 1%, use DOWEX MXA-1, use the 0.2mol/LNaCl eluant solution, obtain the pure product of water-soluble yeast beta-dextran.The beta-glucan content 96% of product, yield 1.2%.
Embodiment 4
Add the 6L pure water in the 200g yeast powder, 121 ℃ are extracted 2h, the centrifugal 10min of 4000r/min, and collecting precipitation I, clear liquid concentrates, alcohol precipitation, drying obtains the yeast mannosans; The potassium hydroxide solution that in precipitation I, adds 4L 4% then, 80 ℃ of stirring reaction 2h, the centrifugal 20min of 8000r/min collects alkali and carries precipitation III.Clear liquid is neutralized to pH6.0 with 8% (w/v) hydrochloric acid, to be cooled after room temperature, the centrifugal 10min of 8000r/min, the clear liquid ultrafiltration and concentration be original volume 1/6 after, add 95% ethanol and make determining alcohol reach 75%, alcohol precipitation obtains water-soluble yeast beta-dextran after the precipitation drying, the beta-glucan content 86% of product, yield 12%.Alkali is carried among the precipitation III and is added 2L water, regulates pH5.0, adds neutral protease 2g, 42 ℃ of enzymolysis 8h, and collecting precipitation, drying obtains non-water-soluble yeast beta-dextran.
Water-soluble yeast beta-dextran redissolves, be mixed with 4% solution, with 0.22um hollow fiber film assembly micro-filtration, with the ultrafiltration of 6KD hollow fiber film assembly, regulate about concentrated solution concentration to 0.5%, with DEAE-Sepharose FF column chromatography purifying, use the 0.2mol/LNaCl eluant solution, obtain the pure product of water-soluble yeast beta-dextran.The beta-glucan content 98% of product, yield 1.5%.
Add the 2L pure water in the 2000mL fresh yeast, 60 ℃ are extracted 1h, the centrifugal 10min of 4000r/min, and collecting precipitation I, clear liquid concentrates, alcohol precipitation, drying obtains the yeast mannosans; The sodium hydroxide that in precipitation I, adds 6L6% then, 80 ℃ of stirring reaction 0.5h, the centrifugal 20min of 5000r/min collects alkali and carries precipitation III.Clear liquid is neutralized to pH7.0 with 12% (w/v) acetate, to be cooled after room temperature, the centrifugal 15min of 5000r/min, the clear liquid ultrafiltration and concentration be original volume 1/5 after, add dehydrated alcohol and make determining alcohol reach 85%, alcohol precipitation obtains water-soluble yeast beta-dextran after the precipitation drying, the beta-glucan content 80% of product, yield 18%.Alkali is carried among the precipitation III and is added 1L water, regulates pH8.5, adds papoid 1g, 55 ℃ of enzymolysis 4h, and collecting precipitation, drying obtains non-water-soluble yeast beta-dextran.
Water-soluble yeast beta-dextran redissolves, be mixed with 3% solution, with 0.22um hollow fiber film assembly micro-filtration, with the ultrafiltration of 6KD hollow fiber film assembly, regulate about concentrated solution concentration to 1%, with DEAE-Sephacel column chromatography purifying, use the 0.4mol/LNaCl eluant solution, obtain the pure product of water-soluble yeast beta-dextran.The beta-glucan content 96% of product, yield 1.8%.
The test embodiment
Experimental example 1: carbon granule clearance test
Test principle: the intravital phagocytic cell of flesh has large and small two kinds.Microphage is the neutrophil leucocyte in the peripheral blood.Macrophagocyte is the scavenger cell in monocyte in the blood and multiple organ, the tissue, and both constitute mononuclear phygocyte system.
Scavenger cell has very strong phagocytic function to particulate antigen, after injecting carbon granules (india ink) in the human body, calculating is engulfed coefficient, is cleaned up index, and chest spleen index, compare with positive control and negative control, can whether can strengthen the phagocytic activity of scavenger cell by verification sample, thereby judge whether it has the function of enhancing body immunological competence.
Test method:
Get Kunming kind small white mouse, random packet, gastric infusion (the water-soluble yeast beta-dextran product that makes among the embodiment 1) is 10 days continuously.Last administration is after 1 hour, accurately inject the india ink 0.1mL/10g that uses 5 times of dilutions of physiological saline by the tail vein respectively,, get blood 20 μ l from the eye socket rear vein beard respectively in injecting prepared Chinese ink 30 seconds and 6 minutes, be blown into immediately among the 0.1% sodium carbonate solution 2mL, fully mixing.Get blood and finish and be dissolved in 2mL0.1% sodium carbonate solution school zero, in 657nm place mensuration optical density(OD) (OD), do blank with 0.1% sodium carbonate solution with 752 type ultraviolet spectrophotometers with 20 μ l normal mouse blood.The dissection mouse is got liver and spleen is weighed, and index K value is cleaned up in calculating and the liver spleen is engulfed coefficient a value, thymus gland and index and spleen index.Data are learned processing by statistics, the results are shown in Table 1 and table 2.
Table 1 mouse macrophage phagocytosis test result
(n＝10，x±s)
Annotate:
*Compare with the physiological saline group, the p value is less than 0.05.Positive control is a Content of Tablet of Levamisole Hydrochloride, is mixed with 10mg/mL solution with preceding with physiological saline.
Table 2 mouse immune organ thymus gland, spleen index test-results
Above test-results shows that the water-soluble yeast beta-dextran sample has the function that the exponential sum phagocytic index is cleaned up in raising, can improve thymus index and index and spleen index simultaneously.
Experimental example 2: antibody generates test
Test method:
Get Kunming kind small white mouse, grouping immediately, gastric infusion (the water-soluble yeast beta-dextran product that makes among the embodiment 1), continuous 9 days, second day mouse peritoneal injection 10%SRBC0.2mL.To produce anti-SRBC antibody.Eye socket venous plexus blood sampling in the 8th day is placed 1h under the room temperature, the centrifugal 10min of 2000r/min, and separation of serum is through 56 ℃ of water-bath 30min deactivation.Separation of serum is done 100 doubling dilutions with physiological saline, get the 1mL dilute serum and add 1%SRBC0.1mL, be diluted to 10% complement 0.5mL with Hanks liquid and mix, 37 ℃ of water-bath 30min, 0 ℃ of termination reaction.Centrifugal, get supernatant liquor and 721 spectrophotometer 540nm place colorimetrics, do the blank zeroing with increase serum not.
Data are learned processing by statistics, the results are shown in Table 3.
Table 3 mouse antibodies generates test-results
Normal mouse is subjected to can produce hemolysin after the SRBC immunity, and this antibody with SRBC, complement incubation, can make the SRBC dissolving external, disengages oxyphorase, and solution is taken on a red color.Measure the optical density(OD) in the supernatant liquor, can judge the quantity that antibody generates in the serum indirectly, optical density(OD) is big more, and it is many more to illustrate that antibody generates.The physiological saline control tube does not then have haemolysis.
The experimental result of table 3 shows that the absorbancy of each dosage group of water-soluble yeast beta-dextran sample all is higher than positive controls, and along with the increase of dosage, optical density(OD) is in rising trend, and the prompting water-soluble yeast beta-dextran has increases the function that mouse antibodies generates.
EXPERIMENTAL EXAMPLE 3: finished product detection
The water-soluble yeast beta-dextran that embodiment 1.2.3 is made carries out thin-layer chromatography and detects, its result as shown in Figure 2:
Point sample is after after developping agent launches, embodiment 1 sample, embodiment 2 samples, embodiment 3 samples all and the glucose sample on same position, illustrate that the polysaccharide that obtained is made up of a kind of monose of glucose, be dextran;
With the infrared detection of carrying out of the water-soluble yeast beta-dextran of embodiment 1 preparation, its infrared spectrogram as shown in Figure 3:
3319-3329cm among the figure
-1Near strong and wide absorption peak is arranged, for the stretching vibration of-O-H key absorbs, be the intermolecular and intramolecular hydrogen bond that O-H forms on the polysaccharide; 2922cm
-1Near be absorbed as saturated C-H stretching vibration signal, medium tenacity; 1658cm
-1Near the characteristic absorbance that is absorbed as acid amides I, 1531cm
-1Near the characteristic absorbance that is absorbed as acid amides II, 1250-1425cm
-1Near the characteristic absorbance that is absorbed as acid amides III a little less than the absorption signal, illustrates and wherein contains a small amount of protein ingredient, is speculated as carbohydrate-binding protein; 1039,1076 and 1157cm
-1Near strong and wide absorption peak is arranged, be the pyranose ring charateristic avsorption band, be the nonsymmetrical vibration peak of its glycosidic link C-O-C, signal is strong, is the typical polysaccharide charateristic avsorption band; 889cm
-1Near obvious absorption is arranged and 850cm
-1No absorption peak shows that its polysaccharide structures is that β-glycosidic link connects but not α-glycosidic link connection, and forming monose is β-D-Glucopyranose; At 770cm
-1Neighbouring is the symmetric vibration peak of pyranose ring C-O-C, a little less than the signal.Contain strong C-O-C nonsymmetrical vibration peak and weak C-O-C symmetric vibration peak simultaneously, show outside its polysaccharide main chain and contain side chain.In addition, at 875cm
-1And 810cm
-1Near no absorption peak shows not contain mannopyranose, gala pyranose.
In conjunction with the constitutional features of Fig. 2 and Fig. 3 demonstration, illustrate that the water-soluble polysaccharide that application preparation method of the present invention is obtained is β-1 from yeast, the 3-D-dextran.
Claims (10)
1. water-soluble yeast beta-dextran, it is characterized in that: described yeast beta-dextran is that raw material makes with complete yeast thalline, and its weight-average molecular weight is 8-20 ten thousand Dalton, is to be made by following preparation method: comprise hot-water cure, alkaline purification:
Wherein said alkaline purification process is:
(1) in hot-water cure gained throw out I, adds alkaline solution, the alkaline solution volume that adds is 2-30 times of precipitation I volume, alkaline concentration is 3-10% (w/v), treatment temp is 60-85 ℃, stirring reaction time 0.5-2h, 3000-12000r/min is centrifugal, collects alkali and carries throw out III, obtains clear liquid;
(2) the gained clear liquid neutralizes with acid behind the alkaline extraction, the pH value scope of neutralization back clear liquid is 5.0-8.0,3000-12000r/min is centrifugal, centrifugal clear liquid is concentrated into the 1/3-1/15 of original volume, the ethanol that adds 80-100% (v/v), make alcohol concn reach 60-80% (v/v) and carry out alcohol and analyse, alcohol is analysed among the gained precipitation II and is promptly contained described water-soluble yeast beta-dextran.
2. the preparation method of water-soluble yeast beta-dextran according to claim 1 comprises hot-water cure, alkaline purification, it is characterized in that:
Described alkaline purification process is:
(1) in hot-water cure gained throw out I, adds alkaline solution, the alkaline solution volume that adds is 2-30 times of precipitation I volume, alkaline concentration is 3-10% (w/v), treatment temp is 60-85 ℃, stirring reaction time 0.5-2h, 3000-12000r/min is centrifugal, collects alkali and carries throw out III, obtains clear liquid;
(2) the gained clear liquid neutralizes with acid behind the alkaline extraction, the pH value scope of neutralization back clear liquid is 5.0-8.0,3000-12000r/min is centrifugal, centrifugal clear liquid is concentrated into the 1/3-1/15 of original volume, the ethanol that adds 80-100% (v/v), make alcohol concn reach 60-80% (v/v) and carry out alcohol and analyse, alcohol is analysed among the gained precipitation II and is promptly contained described water-soluble yeast beta-dextran.
3. preparation method according to claim 2 is characterized in that:
Described alkaline purification process is:
(1) add alkaline solution in hot-water cure gained throw out I, the alkaline solution volume of adding is 10 times of precipitation I volume, and alkaline concentration is 4%-10% (w/v), treatment temp is 60-70 ℃, stirring reaction time 1h, and 3000-12000r/min is centrifugal, collect alkali and carry throw out III, obtain clear liquid;
(2) the gained clear liquid neutralizes with acid behind the alkaline extraction, the pH value scope of neutralization back clear liquid is 5.0-8.0,3000-12000r/min is centrifugal, centrifugal clear liquid is concentrated into 1/5 of original volume, the ethanol that adds 80-100% (v/v), make alcohol concn reach 65-75% and carry out alcohol and analyse, alcohol is analysed among the gained precipitation II and is promptly contained described water-soluble yeast beta-dextran.
4. according to claim 2 or 3 described preparation methods, it is characterized in that: the alkali that described alkaline purification process is used is sodium hydroxide or potassium hydroxide or yellow soda ash or its mixture.
5. according to claim 2 or 3 described preparation methods, it is characterized in that: the acid that described alkaline purification process is used is citric acid or acetate or hydrochloric acid or sulfuric acid or its mixture.
6. according to claim 2 or 3 described preparation methods, it is characterized in that: the described precipitation that contains described water-soluble yeast beta-dextran, after redissolving, separates through column chromatography again water, desalination, drying obtains the high purity water-soluble yeast beta-dextran.
7. preparation method according to claim 6 is characterized in that: described column chromatography medium is DOWEXMXA-1 or Q-Sepharose FF or DEAE-Sephacel or DEAE-Sepharose FF.
8. according to claim 2 or 3 described preparation methods, it is characterized in that: the alkali of gained behind the alkaline extraction is carried throw out III carry out the enzyme processing again, its concrete steps are as follows: carry the water that adds 5-20 times of volume among the throw out III to alkali and make it suspendible, regulate pH4.0-9.0, add proteolytic enzyme, make the proteolytic enzyme total amount reach the 0.02-0.2% (W/V) of suspension total amount, be warmed up to 35-60 ℃, reaction times 2-24h obtains water-insoluble yeast beta-dextran.
9. preparation method according to claim 8 is characterized in that: described proteolytic enzyme is neutral protease or Sumizyme MP or its mixture.
10. preparation method according to claim 8 is characterized in that: after resulting non-water-soluble yeast beta-dextran was handled through enzymolysis, washing, wherein yeast beta-dextran content was 90-99%.
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| UA103283C2 (en) * | 2010-05-14 | 2013-09-25 | Олтек, Инк. | Method for determination of components of cell wall of yeast |
| US20110293784A1 (en) | 2010-05-28 | 2011-12-01 | Anja Wittke | Milk-based nutritional compositions |
| GB201020191D0 (en) * | 2010-11-29 | 2011-01-12 | Biotec Pharmacon Asa | Glucan gels |
| CN102838688B (en) * | 2012-09-12 | 2014-07-30 | 江南大学 | Preparation method of soluble yeast glucan |
| CN103059159B (en) * | 2013-02-01 | 2015-01-07 | 黄河三角洲京博化工研究院有限公司 | Process for extracting mannan from beer yeast powder |
| CN103468765A (en) * | 2013-09-18 | 2013-12-25 | 陕西省微生物研究所 | Biological extracting and purifying method for beer yeast soluble 1,3-beta-D-glucan |
| RU2017118769A (en) * | 2014-10-31 | 2018-11-30 | Винтерсхол Хольдинг Гмбх | METHOD FOR CONCENTRATION OF BETA-GLUCANES |
| CN105001352B (en) * | 2015-07-14 | 2017-04-26 | 青岛海大海洋生物医药销售有限公司 | Beta-1,3/1,6-glucan, preparation method therefor, and application thereof in preparing immune enhancement and anti-tumor medicine and functional food |
| CN106243239B (en) * | 2016-07-29 | 2018-11-27 | 山东大学齐鲁医院 | It is a kind of for improving the soluble small molecule beta-1,3-dextran of hepatitis immunity |
| CN108611385A (en) * | 2016-12-12 | 2018-10-02 | 安琪酵母股份有限公司 | Water-soluble yeast beta-dextran and its preparation method and application |
| CN106755196A (en) * | 2016-12-20 | 2017-05-31 | 广东工业大学 | One kind improves the water miscible method of beta glucan |
| CN111011609A (en) * | 2019-12-24 | 2020-04-17 | 仙乐健康科技(安徽)有限公司 | A composition for enhancing immunity of mammals |
| CN114304353B (en) * | 2021-12-30 | 2023-09-19 | 艾苛密（上海）健康科技股份有限公司 | Beta-glucan candy tablet with blood glucose and blood pressure reducing health care function and preparation method thereof |
| CN114343045B (en) * | 2022-01-20 | 2023-11-17 | 河南省寓生堂食养科技有限公司 | Health care tabletting candy and preparation method thereof |
| CN115590787B (en) * | 2022-10-13 | 2023-12-26 | 湖南天根乐微君科技有限公司 | Composition for acne removal and skin repair as well as preparation method and application thereof |
| CN115975069A (en) * | 2023-01-04 | 2023-04-18 | 河南大学 | High-purity water-soluble yeast beta-glucan and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5532223A (en) * | 1989-09-08 | 1996-07-02 | Alpha-Beta Technology, Inc. | Use of aqueous soluble glucan preparations to stimulate platelet production |
| CN101117357A (en) * | 2007-09-17 | 2008-02-06 | 中国农业大学 | Method for preparing water-soluble beta-1,3/1,6-dextran |
-
2008
- 2008-09-17 CN CN2008101402143A patent/CN101353383B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5532223A (en) * | 1989-09-08 | 1996-07-02 | Alpha-Beta Technology, Inc. | Use of aqueous soluble glucan preparations to stimulate platelet production |
| CN101117357A (en) * | 2007-09-17 | 2008-02-06 | 中国农业大学 | Method for preparing water-soluble beta-1,3/1,6-dextran |
Non-Patent Citations (2)
| Title |
|---|
| David L. Williams,et al..A method for the solubilization of a (1→3)-β-D-glucan isolated from Saccharomyces cerevisiae.Carbohydrate Research.1991,219 * |
| 李春林.灰树花水溶性β-葡聚糖的制备和结构研究.中国优秀博硕士学位论文全文数据库(硕士)工程科技I辑.2005,(1), * |
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