CN104292359A - Method for extracting soybean polysaccharide from soybean meal with biological enzyme method - Google Patents
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Abstract
The invention belongs to the technical field of extraction of polysaccharides and particularly relates to a method for extracting a soybean polysaccharide from soybean meal with a biological enzyme method. The method for extracting the soybean polysaccharide from the soybean meal with the biological enzyme method comprises steps as follows: (1) pretreatment of raw materials; (2) enzymolysis; (3) water-insoluble polysaccharide conversion; (4) centrifugal separation; (5) soluble polysaccharide extraction. Adopted enzymes are mild in action condition, various enzymes are adopted for performing enzymolysis on fat and cellulose in the soybean meal to extract the polysaccharide in the fat and the cellulose, the utilization rate of the soybean meal is increased, and the soybean polysaccharide obtained with the method is high in yield and 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 soybean polysaccharide from soybean meal.
background technology
The nutritive ingredient of soybean comprehensively and rich content, this makes the soyabean processing industry of China and even countries in the world all in the ascendant, and along with the further investigation to soybean function composition, the comprehensive development and utilization of soybean is worth the concern being also more and more subject to countries in the world.Soybean protein, soybean polypeptide, the functional components such as soybean isoflavones, soybean phospholipid and large saponin are also obtained for more deep research.But as the by product that soybean protein processing industry is maximum---bean dregs, but within the quite a long time ignore by people, its research is also only rested in general application.The bad process of some enterprises to bean dregs is returned environment and is caused very large mud.Containing abundant albumen in soybean, fat, food fibre, also containing a large amount of Mierocrystalline cellulose, this is good sugar source, utilizes these Mierocrystalline celluloses can obtain the water soluble soybean polysaccharide of a large amount of characteristic goods.Water soluble soybean polysaccharide mainly with the by product soybean-drag fiber of the process for processing such as soybean protein isolate, bean curd and bean curd stick for main raw material, form through the techniques such as pre-treatment, enzymolysis (cellulase, hemicellulase, proteolytic enzyme etc.), separation, decolouring, sterilizing, drying are refining.Soybean polysaccharide be white to pale yellow powder, solution viscosity is low, mouthfeel good.Soybean polysaccharide is except being a kind of excellent water-soluble dietary fibre, and a kind of high performance protein drinks emulsion stabilizer especially, especially has more unique stable emulsifying protein capability under sour environment.Soybean polysaccharide viscosity compared with other biological polyoses is lower, and has the features such as dispersiveness, stability, emulsifying property and tackyness.Be often used as foodstuff fibre reinforcer in food service industry, hold infusion and food properties activator.In addition, solubility many beans polysaccharide also have regulate blood glucose value and blood lipid, the absorption of promotion harmful intestinal tract material and excretion, anticancer, promote the biologic activity such as mineral absorption usability, in anti-oxidant, antibacterial, antiviral and immunomodulatory etc., also have certain effect.
Just soybean polysaccharide is extracted in the basic conditions as far back as Kawamura and Narasaki in 1961, Morita is extracted soybean polysaccharide in nineteen sixty-five in 100 DEG C of hot water, the people such as Aspinall were extracted polysaccharose substance in 1967 from soybean, and preliminary study has been carried out to its structure, after this reported the extracting method of some soybean polysaccharide abroad successively.But domestic research is in this respect also less, and yield is not high in actual applications, and technics comparing is complicated, and cost is also higher.
No matter be that soluble soybean polysaccharide all extremely has development prospect from world market or from domestic market.The soluble soybean polysaccharide mainly dependence on import that Current Domestic uses, price is higher, to such an extent as to the degree of understanding of domestic many enterprises to soluble soybean polysaccharide is inadequate, therefore research is applicable to the soluble soybean polysaccharide system of China's national situation and not only can solves environmental problem caused by bean dregs and a business difficult problem, can also drive the development of related industries, this is also the growth requirement of soyabean processing.If improve its performance further, just can expand the Application Areas of soluble soybean polysaccharide further, produce huge economic benefit.
Therefore need to improve above-mentioned method, soybean meal can use by research one to greatest extent, extracts the method for soybean polysaccharide contained in soybean meal.
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 soybean polysaccharide from soybean meal.
The method that biological enzyme of the present invention extracts soybean polysaccharide from soybean meal is realized by following technical scheme:
Biological enzyme extracts a method for soybean polysaccharide from soybean meal, comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 2-4%, then micronizing is to 800-1000 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal 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 bromeline again, the addition of bromeline is 0.5-1.2%, hydrolysis temperature is 50 DEG C, pH value 6.5, 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, go out after enzymolysis enzyme 5-10min at 90-95 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 95 DEG C of enzyme 5-10min that go out;
(4) centrifugation
By the soybean meal 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 (5) 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 soybean meal after micronizing.
Preferably, the consumption of lipase is 0.4%.
Preferably, the consumption of cellulase is 0.5%.
Preferably, the addition of bromeline is 0.8%.
Preferably, diastatic addition is 1.0%.
Preferably, a kind of biological enzyme extracts the method for soybean polysaccharide from soybean meal, comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 2-4%, then micronizing to 800 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal and water is 1:9, 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 bromeline again, the addition of bromeline is 0.8%, and hydrolysis temperature is 50 DEG C, pH value 6.5, is placed in Microwave Extraction Equipment by above-mentioned raw material and extracts, and the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h, and go out after enzymolysis enzyme 8min at 90-95 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 95 DEG C of enzyme 6min that go out;
(4) centrifugation
By the soybean meal 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 (5) 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 soybean meal after micronizing.
In method of the present invention, first soybean meal is pulverized, adopt the method for micronizing, instead of common grinding mode, because after micronizing, the particle diameter of soybean meal is less, be conducive in follow-up enzyme digestion reaction process, various component contacts with enzyme, makes enzyme better act on each component;
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 soybean meal, 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 soybean meal, 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 soybean meal 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, and 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 soybean meal 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.
Beneficial effect of the present invention is, adopts enzyme action condition gentle, adopts various different enzyme by the fat in soybean meal, cellulase hydrolysis, from soybean meal, extract polysaccharide simultaneously, utilize soybean meal 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 soybean polysaccharide from soybean meal, comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 3%, then micronizing to 800 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal and water is 1:9, 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 bromeline again, the addition of bromeline is 0.8%, and hydrolysis temperature is 50 DEG C, pH value 6.5, is placed in Microwave Extraction Equipment by above-mentioned raw material and extracts, and the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h, and go out after enzymolysis enzyme 8min at 90-95 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1%, hydrolysis temperature 40 DEG C, pH value 5, and after enzymolysis 1.8h, 95 go out enzyme 6min;
(4) centrifugation
By the soybean meal 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 soybean meal after micronizing.
The yield of polysaccharide is: 13.98%, and purity is 97.4%.
Embodiment 2
Biological enzyme extracts a method for soybean polysaccharide from soybean meal, comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 2%, then micronizing to 800 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal 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;
Add bromeline again, the addition of bromeline is 0.5%, and hydrolysis temperature is 50 DEG C, pH value 6.5, is placed in Microwave Extraction Equipment by above-mentioned raw material and extracts, and the power of microwave is 400W, frequency is 800MHZ, enzymolysis 0.1h, at 95 times enzyme 5min that go out after enzymolysis;
(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 soybean meal 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 soybean meal after micronizing.
The extraction yield of polysaccharide is: 1 1.85%, and purity is 96.8%.
Embodiment 3
Biological enzyme extracts a method for soybean polysaccharide from soybean meal, comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 4%, then micronizing to 800 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal 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 0.1-0.5h;
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 bromeline again, the addition of bromeline is 1.2%, and hydrolysis temperature is 50 DEG C, pH value 6.5, is placed in Microwave Extraction Equipment by above-mentioned raw material and extracts, and the power of microwave is 600W, frequency is 800MHZ, enzymolysis 0.5h, and go out after enzymolysis enzyme 10min at 95 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 soybean meal 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 soybean meal after micronizing.
The yield of polysaccharide is: 12.54%, and purity is 96.5%.
Claims (6)
1. biological enzyme extracts a method for soybean polysaccharide from soybean meal, comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 2-4%, then micronizing is to 800-1000 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal 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 bromeline again, the addition of bromeline is 0.5-1.2%, hydrolysis temperature is 50 DEG C, pH value 6.5, 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, go out after enzymolysis enzyme 5-10min at 90-95 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 90-95 DEG C;
(4) centrifugation
By the soybean meal 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 soybean meal after micronizing.
2. a kind of biological enzyme extracts the method for soybean polysaccharide from soybean meal 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 soybean polysaccharide from soybean meal, 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 soybean polysaccharide from soybean meal, it is characterized in that, the addition of described bromeline is 0.8%.
5. a kind of biological enzyme as claimed in claim 1 extracts the method for soybean polysaccharide from soybean meal, it is characterized in that, described diastatic addition is 1.0%.
6. a kind of biological enzyme according to any one of claim 1-4 extracts the method for soybean polysaccharide from soybean meal, it is characterized in that, described method comprises following step:
(1) raw materials pretreatment
Soybean meal drying to its moisture content is 2-4%, then micronizing to 800 order;
(2) enzymolysis
By the soybean meal after pulverizing, add water mixing, the part by weight of soybean meal and water is 1:9, 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 bromeline again, the addition of bromeline is 0.8%, and hydrolysis temperature is 50 DEG C, pH value 6.5, is placed in Microwave Extraction Equipment by above-mentioned raw material and extracts, and the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.2h, and go out after enzymolysis enzyme 8min at 95 DEG C;
(3) water-insoluble polysaccharide conversion
Diastatic addition is 1%, hydrolysis temperature 40 DEG C, pH value 5, after enzymolysis 1.8h, and 95 DEG C of enzyme 6min that go out;
(4) centrifugation
By the soybean meal 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 soybean meal after micronizing.
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