CN104387484A - Method for extracting rapeseed polysaccharide from rape seed cake by bioenzyme process - Google Patents
Method for extracting rapeseed polysaccharide from rape seed cake by bioenzyme process Download PDFInfo
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- CN104387484A CN104387484A CN201410628551.2A CN201410628551A CN104387484A CN 104387484 A CN104387484 A CN 104387484A CN 201410628551 A CN201410628551 A CN 201410628551A CN 104387484 A CN104387484 A CN 104387484A
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Abstract
The invention belongs to the technical field of extraction, and particularly relates to a method for extracting rapeseed polysaccharide from rape seed cake by a bioenzyme process. The method comprises the following steps: (1) raw material pretreatment; (2) enzymolysis; (3) non-water-soluble polysaccharide conversion; (4) centrifugal separation; and (5) soluble polysaccharide extraction. Under mild enzyme conditions, different enzymes are adopted to decompose fat and cellulose in the rape seed cake so as to extract the polysaccharide in the rape seed cake, thereby enhancing the utilization ratio of the rape seed cake. The rapeseed polysaccharide obtained by the method has the advantages of high yield and high purity.
Description
Technical field
The invention belongs to technical field of polysaccharide extraction, be specifically related to a kind of method that biological enzyme extracts rapeseed polysaccharides from dregs of rapeseed cake.
background technology
In order to improve economic benefit and the social benefit of rapeseed product, reply dregs of rapeseed cake fully utilizes, in dregs of rapeseed cake, available composition mainly contains protein, the compositions such as carbohydrate, and wherein sulphur glucoside, sinapinic acid, phytic acid, polyphenolic substance and polyose composition, as non-starch polysaccharide, be regarded as anti-nutrition component always, think that their High Temperature Pres cause the variability of protein when squeezing, the biological value of protein is reduced greatly, thus do not play a positive role in actual production, existing Preliminary Study shows at present, rapeseed polysaccharides is as the one of vegetable polysaccharides, oxidation resistant effect can be played by other reductibility composition in the restitutive protection organism of self.
The traditional extraction process of vegetable polysaccharides has hot water extraction, diluted acid lixiviate, diluted alkaline lixiviate, along with the progress of technology, introduces some new technologies in recent years, novel method in extraction process, as Enzymatic Extraction, and microwave extraction, ultrasonic extraction etc.
If can by the polysaccharide extraction in dregs of rapeseed cake out and make full use of, good economic benefit and social benefit will be brought, but it is still rare for the report of rapeseed polysaccharides at present, therefore, select a kind of best approach of simple, fast and efficient extraction rapeseed polysaccharides, tool is of great significance.
Extracting method main is at present hot water extraction, and the polysaccharide larger due to molecular mass and acidic polysaccharose in the hot water solubleness are less, and adopt alkaline extraction efficiency higher, water extraction can be difficult to the acidic polysaccharose that separates and mucopolysaccharide extracts.Yan Fengwei etc. are while dregs of rapeseed cake comprehensively extracts polyphenol, phytic acid and protein, be separated from polyphenol extracting solution again and obtain rapeseed polysaccharides crude product, and chemically simulated system, subcellular organelle, tissue and animal body level have rated the antioxygenation of rapeseed polysaccharides crude product, prove that rapeseed polysaccharides all has stronger antioxygenation in vivo and in vitro.By free-radical theory, the exploitation of rapeseed polysaccharides open one and have valuable direction, and substantially increase the comprehensive utilization ratio of dregs of rapeseed cake.Adopt the method action condition of alkaline extraction strong, its extraction yield is not high, about 7.6%.
Therefore need to improve above-mentioned method, dregs of rapeseed cake can use by research one to greatest extent, such as the efficient method extracted by rapeseed polysaccharides contained in dregs of rapeseed cake, and extraction yield is high.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of method that yield and the higher biological enzyme of purity extract rapeseed polysaccharides from dregs of rapeseed cake.
The method that biological enzyme of the present invention extracts rapeseed polysaccharides from dregs of rapeseed cake is realized by following technical scheme:
Biological enzyme extracts a method for rapeseed polysaccharides from dregs of rapeseed cake, comprises following step:
(1) raw materials pretreatment
Dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 2-4% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 80-200 order;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:6-12, and adds lipase, the addition 0.3-0.5% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 400-600W, frequency is 800MHZ, enzymolysis 0.1-0.5h;
Add cellulase again, the addition of cellulase is 0.4-0.6%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, and the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h;
Add aspartic protease and papoid again, the addition of aspartic protease is 0.5-1.2%, the addition of papoid is 0.3-0.6%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 400-600W, frequency is 800MHZ, enzymolysis 0.1-0.5h, and go out after enzymolysis enzyme 5-10min at 100-105 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1.0-1.2%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.5-2.0h, and the 100-105 DEG C of enzyme 5-10min that goes out;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 10-20 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 60-70 DEG C, being concentrated into its concentration is 60-70%, concentrated solution adds in 1:4 ratio that 95% alcohol settling leaves standstill 10-20 hour, centrifugal 10-20 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 4-6%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
Preferably, the consumption of lipase is 0.4%.
Preferably, the consumption of cellulase is 0.5%.
Preferably, diastatic addition is 1.0%.
Preferably, a kind of biological enzyme extracts the method for rapeseed polysaccharides from dregs of rapeseed cake, comprises following step:
(1) raw materials pretreatment
Dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 2-4% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 80-200 order;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:8, and adds lipase, the addition 0.4% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h;
Add cellulase again, the addition of cellulase be 0.5%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 500W, and frequency is 800MHZ, enzymolysis 0.2h;
Add aspartic protease and papoid again, the addition of aspartic protease is 0.8%, the addition of papoid is 0.5%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h, and go out after enzymolysis enzyme 8min at 100 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 100 DEG C of enzyme 6min that go out;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% alcohol settling in 1:4 ratio and leaves standstill 15 hours, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
In the present invention, by rapeseed meal by supercritical carbon dioxide extraction, extract the grease in dregs of rapeseed cake further, not only obtain grease, and be conducive to the extraction of follow-up rapeseed polysaccharides.
Enzymolysis fat, fiber order are also innovations of the present invention, and due to by after fatty enzymolysis, grease can not be wrapped in outside dregs of rapeseed cake, thus are conducive to various composition and contact with enzyme, thus the efficiency of raising enzymolysis; Therefore, the fatty enzymolysis Mierocrystalline cellulose again of first enzymolysis is conducive to the removal of fat and fiber;
Generally adopt the mode of organic solvent of adding to the treatment process of fat in usual way, although organic solvent is also comparatively thorough to the removal of grease, but the condition of its effect is stronger, and follow-up removal process is also more complicated, also easily causes the environmentally hazardous problem of organic solvent in addition; Organic solvent is adopted to remove its cost of grease in addition also higher;
The present invention adopts enzyme to act on to remain on the grease in dregs of rapeseed cake, and action condition is gentle, and also comparatively thorough to the removal of grease, and there is not the problem that follow-up organic solvent reclaims and bring environmental pollution;
Add cellulase again and the cellulose degradation in dregs of rapeseed cake is generated water-soluble glucose;
After the polysaccharide enzymolysis of non-solubility is soluble polysaccharide, carbohydrate is soluble in water, then gets supernatant liquor, filters, concentrated, alcohol precipitation, by Polyose extraction wherein out.
The selection of the kind of enzyme of the present invention and the selection of consumption and enzymolysis order is not accidental, but contriver has paid performing creative labour obtains, the adjustment of enzyme and various ratio all can affect the extraction effect of final polysaccharide, only have adopt the present invention to enzyme and the corresponding proportion of enzyme dregs of rapeseed cake is processed, and carry out according to order of the present invention, just can obtain result of the present invention, enzyme be carried out replace or the replacement of enzyme-added order, all can not get maximum polysaccharide extract rate.
Therefore, adopt the method for freezing and pulverizing to extracting the polysaccharide obtained in the present invention, the quality maintaining polysaccharide is unaffected.
Beneficial effect of the present invention is, adopt enzyme action condition gentle, adopt various different enzyme by the fat in dregs of rapeseed cake, cellulase hydrolysis, from dregs of rapeseed cake, extract polysaccharide simultaneously, utilize dregs of rapeseed cake to greatest extent, and the polysaccharide adopting method of the present invention to obtain, its yield and purity high.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described, so that those skilled in the art more understands the present invention, but does not therefore limit the present invention.
Following detection method, if no special instructions, adopts phend-sulphuric acid to survey the content of polysaccharide.
Embodiment 1
Biological enzyme extracts a method for rapeseed polysaccharides from dregs of rapeseed cake, comprises following step:
(1) raw materials pretreatment
Dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 3% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 100 orders;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:8, and adds lipase, the addition 0.4% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h;
Add cellulase again, the addition of cellulase be 0.5%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 500W, and frequency is 800MHZ, enzymolysis 0.2h;
Add aspartic protease and papoid again, the addition of aspartic protease is 0.8%, the addition of papoid is 0.5%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h, and go out after enzymolysis enzyme 8min at 100 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 100 DEG C of enzyme 6min that go out;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% alcohol settling in 1:4 ratio and leaves standstill 15 hours, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
The content adopting phend-sulphuric acid to record wherein polysaccharide is about 6.72%.
Embodiment 2
Biological enzyme extracts a method for rapeseed polysaccharides from dregs of rapeseed cake, comprises following step:
(1) raw materials pretreatment
Dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 2% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 80 orders;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:6, and adds lipase, the addition 0.3% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 400W, frequency is 800MHZ, enzymolysis 0.1h;
Add cellulase again, the addition of cellulase be 0.4%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 400W, and frequency is 800MHZ, enzymolysis 0.1-0.5h;
Add aspartic protease and papoid again, the addition of aspartic protease is 0.5%, the addition of papoid is 0.3%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 400W, frequency is 800MHZ, enzymolysis 0.1h, and go out after enzymolysis enzyme 5min at 100 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 0.8%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 100 DEG C of enzyme 6min that go out;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% alcohol settling in 1:4 ratio and leaves standstill 15 hours, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
The polysaccharide content adopting phend-sulphuric acid to record gained is 6.06%.
Embodiment 3
Biological enzyme extracts a method for rapeseed polysaccharides from dregs of rapeseed cake, comprises following step:
(1) raw materials pretreatment
Dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 4% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 200 orders;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:12, and adds lipase, the addition 0.5% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 600W, frequency is 800MHZ, enzymolysis 0.5h;
Add cellulase again, the addition of cellulase be 0.6%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 600W, and frequency is 800MHZ, enzymolysis 0.5h;
Add aspartic protease and papoid again, the addition of aspartic protease is 1.2%, the addition of papoid is 0.6%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 600W, frequency is 800MHZ, enzymolysis 0.5h, and go out after enzymolysis enzyme 5-10min at 100 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1.2%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 100 DEG C of enzyme 6min that go out;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% alcohol settling in 1:4 ratio and leaves standstill 15 hours, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
The content adopting phend-sulphuric acid to record sugar in the polysaccharide of gained is 6.21%.
Claims (5)
1. biological enzyme extracts a method for rapeseed polysaccharides from dregs of rapeseed cake, comprises following step:
(1) raw materials pretreatment
A. dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 2-4% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 80-200 order;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:6-12, and adds lipase, the addition 0.3-0.5% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 400-600W, frequency is 800MHZ, enzymolysis 0.1-0.5h;
Add cellulase again, the addition of cellulase is 0.4-0.6%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, and the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h;
Add aspartic protease and papoid again, the addition of aspartic protease is 0.5-1.2%, the addition of papoid is 0.3-0.6%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 400-600W, frequency is 800MHZ, enzymolysis 0.1-0.5h, and go out after enzymolysis enzyme 5-10min at 100-105 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 0.8-1.2%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.5-2.0h, and the 100-105 DEG C of enzyme 5-10min that goes out; Above-mentioned raw material is placed in Microwave Extraction Equipment extract, the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h, and go out after enzymolysis enzyme 5-10min at 100-105 DEG C;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 10-20 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 60-70 DEG C, being concentrated into its concentration is 60-70%, concentrated solution adds in 1:4 ratio that 95% alcohol settling leaves standstill 10-20 hour, centrifugal 10-20 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 4-6%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
2. a kind of biological enzyme extracts the method for rapeseed polysaccharides from dregs of rapeseed cake as claimed in claim 1, it is characterized in that, the consumption of described lipase is 0.4%.
3. a kind of biological enzyme as claimed in claim 1 extracts the method for rapeseed polysaccharides from dregs of rapeseed cake, it is characterized in that, the consumption of described cellulase is 0.5%.
4. a kind of biological enzyme as claimed in claim 1 extracts the method for rapeseed polysaccharides from dregs of rapeseed cake, it is characterized in that, described diastatic addition is 1.0%.
5. a kind of biological enzyme according to any one of claim 1-4 extracts the method for rapeseed polysaccharides from dregs of rapeseed cake, it is characterized in that, described method comprises following step:
(1) raw materials pretreatment
Dregs of rapeseed cake pulverizer is crushed to 40 orders, dress extraction kettle, and described dregs of rapeseed cake is that peony seeds benevolence just squeezes rear remaining residual dregs of rice slag;
B. use food-class CO 2, its purity is 99.99%, and flow is 12L/ hour, and setting extracting pressure is 20MPa, temperature
Degree is 25 DEG C, and separation condition is: separator I pressure 10KPa, temperature 50 C, separator II pressure 4KPa, temperature 40 DEG C, extraction time 0.5 hour;
C. extract complete, collect extract, obtain peony seed oil;
D. getting residue processed remaining in step c is 20% to its moisture content, obtains dregs of rapeseed cake;
Be 2-4% by the dregs of rapeseed cake drying in steps d to its moisture content, then be crushed to 80-200 order;
(2) enzymolysis
By the dregs of rapeseed cake after pulverizing, add water mixing, the part by weight of dregs of rapeseed cake and water is 1:8, and adds lipase, the addition 0.4% of lipase, hydrolysis temperature 45 DEG C, adjust pH 5.5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h;
Add cellulase again, the addition of cellulase be 0.5%, hydrolysis temperature 40 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment and extracts, the power of microwave is 500W, and frequency is 800MHZ, enzymolysis 0.2h;
Add aspartic protease and papoid again, the addition of aspartic protease is 0.8%, the addition of papoid is 0.5%, hydrolysis temperature is 45 DEG C, and above-mentioned raw material is placed in Microwave Extraction Equipment and extracts by pH value 5.0, the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h, and go out after enzymolysis enzyme 8min at 100 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 100 DEG C of enzyme 6min that go out;
(4) centrifugation
By the dregs of rapeseed cake after enzymolysis, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, supernatant liquor and precipitation is collected respectively;
(5) extraction of soluble polysaccharide
The supernatant liquid filtering got in step (4) removes slag, the filtrate obtained after filtration is concentrated at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% alcohol settling in 1:4 ratio and leaves standstill 15 hours, centrifugal 15 min, centrifugal rotational speed is 4500 rpm, and collecting precipitation also cleans 2-3 time repeatedly with ethanol, and lyophilize to moisture content is after 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide;
Above-mentioned enzyme concentration is the weight percent that enzyme accounts for the dregs of rapeseed cake after micronizing.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108936701A (en) * | 2018-08-02 | 2018-12-07 | 句容市惠农油脂有限公司 | A kind of preparation method of vegetable seed meal dietary fiber |
| CN109054988A (en) * | 2018-08-22 | 2018-12-21 | 孟凡志 | A kind of rapeseed oil extraction process |
| CN116076699A (en) * | 2021-11-05 | 2023-05-09 | 安琪生物科技有限公司 | Plant raw material meal enzymolysis product and preparation method and application thereof |
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| CN108936701A (en) * | 2018-08-02 | 2018-12-07 | 句容市惠农油脂有限公司 | A kind of preparation method of vegetable seed meal dietary fiber |
| CN109054988A (en) * | 2018-08-22 | 2018-12-21 | 孟凡志 | A kind of rapeseed oil extraction process |
| CN116076699A (en) * | 2021-11-05 | 2023-05-09 | 安琪生物科技有限公司 | Plant raw material meal enzymolysis product and preparation method and application thereof |
| RU2815687C1 (en) * | 2023-08-23 | 2024-03-20 | федеральное государственное бюджетное образовательное учреждение высшего образования "Российский биотехнологический университет (РОСБИОТЕХ)" | Method of producing complex of polysaccharides from rapeseed cake with coextracted protein |
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