CN103361395A - Solid-state fermentation preparation method for peanut non-starch polysaccharides - Google Patents
Solid-state fermentation preparation method for peanut non-starch polysaccharides Download PDFInfo
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Abstract
The invention discloses a solid-state fermentation preparation method for peanut non-starch polysaccharides. The method comprises a step of uniformly mixing peanut meal and a nutritive salt solution after the peanut meal and the nutritive salt solution are disinfected, and inoculating a strain to perform solid-state fermentation; a step of adding sterile water, extracting in a thermostatic water bath, centrifuging and performing suction filtration; and a step of separating and purifying by using ultrafiltration membranes, adding absolute ethyl alcohol having a volume 4 times of the volume of the permeate liquid, allowing the mixture to stand overnight, centrifuging, precipitating and freeze drying to obtain the peanut non-starch polysaccharides. The peanut non-starch polysaccharides prepared by the method have high yield, high purity, small contents of impurity proteins, and functional activity of antioxidation and blood sugar reduction, and are suitable for industrial production.
Description
Technical field
The present invention relates to the method that solid state fermentation prepares the peanut non-starch polysaccharide, belong to food nutrition active substance manufacture field.
Background technology
2010/2011 annual China peanut pressing quantity thereof is 9,000,000 tons, produces about 4,500,000 tons of peanut meal, and peanut meal contains protein about 50% and about 40% carbohydrate, is a kind of large natural protein and polysaccharide resource.Bacterial classification accessed carry out solid state fermentation in the peanut meal, peanut meal can be used as nutritive substance and utilizes for the microorganism growth breeding, microorganism secretes plurality of enzymes system in growth and development process, comprise proteolytic enzyme, lipase, amylase, saccharifying enzyme, cellulase, phytase etc., these enzymes are small molecular protein, polypeptide and small peptide etc. with the peanut meal proteolysis, be active polysaccharide-non-starch polysaccharide with carbohydrate inversion simultaneously, improve peanut meal nutritive value and physiologically active, improve functional performance.Through different extraction, separation, purification process, the peanut meal after fermentation can obtain the products such as biologically active peptides, non-starch polysaccharide and food brewing special protein base-material, reaches the purpose of peanut meal higher value application.
The peanut non-starch polysaccharide is a kind of light brown, the pulverulent solids of sugar and peanut fragrance slightly, is soluble in hot water.The mixing polysaccharide that this product is comprised of reducing sugar, pentose, uronic acid or glycoprotein, molecular weight have higher physiologically active about 30kD, as removing the effects such as free radical, anti-oxidant, hypoglycemic, reducing blood-fat, raising immunizing power.The hypoglycemic mechanism of peanut non-starch polysaccharide is that reversibility competition Inhibiting α-glucosidase is active, reduces the decomposition rate of starch, sucrose, maltose, reduces the speed of digesting and assimilating of glucose, thus the control blood sugar increasing.The peanut non-starch polysaccharide is directly taken in can incapsulating as protective foods, as the assisting therapy product of diabetes; Also can be added in diabetics's special foods such as Nutritive Rice paste, sugar-free biscuit, as diabetes rehabilitation product and daily nutrition complementary goods; Also can be used as natural food-preservative and be added in bakery, the beverage product, prevent fats oxidn, prolong Food Shelf-life.
At present, the preparation technology of peanut non-starch polysaccharide adopts hot water, ethanolic soln, acid solution or the methods such as alkaline solution lixiviate, enzymic hydrolysis more.There are many problems in these methods, and large such as hot water lixiviate energy consumption, efficient is low; The peanut non-starch polysaccharide volatility of ethanolic soln lixiviate, Product Activity is low; Bronsted lowry acids and bases bronsted lowry solution extraction contaminate environment, product volatility, active low; The used commercial enzyme degree of hydrolysis of enzyme hydrolysis method is difficult to control, and product foreign protein content is high.Therefore, need that a kind of power consumption is low, efficient is high, extraction yield is high, foreign protein content is low, environmental friendliness, peanut non-starch polysaccharide Product Activity height, and the extracting method of the wide peanut non-starch polysaccharide of suitability for industrialized production prospect and market outlook.
Summary of the invention
The object of the invention is to the technical barrier that exists in the above-mentioned technology for solving, a kind of method low, that solid state fermentation that efficient is high, extraction yield is high, foreign protein content is low, environmental friendliness, peanut non-starch polysaccharide Product Activity height and suitability for industrialized production have a extensive future prepares the peanut non-starch polysaccharide that consumes energy is provided.
For achieving the above object, the technical solution adopted in the present invention is: a kind of solid state fermentation prepares the method for peanut non-starch polysaccharide, and its step comprises:
One, solid state fermentation: peanut meal is pulverized, and crosses 120 mesh sieves, gets the lower peanut meal powder of sieve as raw material, and accesses mixed strains, solid state fermentation in constant incubator after the nutrient salt solution sterilization;
Two, lixiviate: add sterilized water in the fermented product, lixiviate in the water bath with thermostatic control vibrator, centrifugal, vacuum filtration;
Three, separation and purification: suction filtration liquid is got the dehydrated alcohol that adds 4 times of volumes through liquid through the ultra-filtration membrane separation and purification, and hold over night in the refrigerator is centrifugal, and the precipitation lyophilize obtains the peanut non-starch polysaccharide.
Preferably, the nutrient salt solution in the described step 1 is to contain 0.55% (NH in the 1L volume
4)
2SO
4, 0.45% KH
2PO
4, 0.9% urea, 1.1% glucose, the condition of sterilization is 121 ℃ of sterilization 15min, the quality of peanut meal powder and nutrient salt solution and volume ratio are 1:0.5～1.75(g/ml), the bacterial classification of access is that cell age is that the aspergillus oryzae of 60～84h, cereuisiae fermentum and the cell age that cell age is 30～40h are the mixed strains of the milk-acid bacteria of 9～15h, the blending ratio of bacterial classification is aspergillus oryzae: cereuisiae fermentum: milk-acid bacteria=1:0.5～1.5:0.5～1.0, the access volume ratio of peanut meal powder and bacterial classification is 1:0.1～0.2(g/ml, 10
8Individual/the ml bacterial classification), solid state fermentation conditions is 25～30 ℃ of lower standing for fermentation 30～40h.
Preferably, fermented product in the described step 2 and the quality of sterilized water and volume ratio are 1:3.5～4.5(g/ml), the water bath with thermostatic control extracting condition is under 25～35 ℃, in the water bath with thermostatic control vibrator, with 125r/min rotating speed cyclotron oscillation mode lixiviate 4～6h, centrifugal condition is the centrifugal 20min of 4500r/min, and vacuum filtration obtains suction filtration liquid under the vacuum tightness 0.1MPa.
Preferably, the ultra-filtration membrane purification condition in the described step 3 is that to select molecular weight cut-off be the polysulfone hollow fibre ultra-filtration membrane of 30kDa, 0.075MPa pressure, 28 ℃ of lower ultrafiltration, centrifugal condition is the centrifugal 20min of 4500r/min, and the lyophilize condition is-55 ℃, vacuum tightness 0.1MPa, dry 48h.
The invention has the beneficial effects as follows: processing step disclosed in this invention power consumption is low, efficient is high, extraction yield up to more than 78%, purity is greater than 97%, foreign protein content is low, environmental friendliness, peanut non-starch polysaccharide Product Activity height and suitability for industrialized production have a extensive future.
Embodiment
Adopt the phenolsulfuric acid method to measure hexose content, orcinol colorimetric method for determining pentose content, carbazole-Sulphuric acid Colorimetry glucuronic acid content, according to hexose, pentose, the uronic acid weight ratio in non-starch polysaccharide, according to formula: non-starch polysaccharide content=hexose content * 0.9+pentose content * 0.88+glucuronic acid content * 0.81 calculates the purity of the non-starch polysaccharide that extracts in the embodiment of the invention.Adopt micro-Kjeldahl to measure the impurity protein content of the non-starch polysaccharide that extracts in the embodiment of the invention.
Embodiment 1
Peanut meal is pulverized, and crosses 120 mesh sieves, and peanut meal powder and the nutrient salt solution got under the sieve (contain 0.55% (NH in the 1L volume
4)
2SO
4, 0.45% KH
2PO
4, 0.9% urea, 1.1% glucose) at 121 ℃ of sterilization 15min, with 1:1.25(g/ml) ratio in the peanut meal powder, add nutrient salt solution, mix, with 1:0.15(g/ml, 10
8Individual/the ml bacterial classification) ratio access cell age in the peanut meal powder be that the aspergillus oryzae of 72h, cereuisiae fermentum and the cell age that cell age is 36h are the mixed strains of the milk-acid bacteria of 12h, the blending ratio of bacterial classification is aspergillus oryzae: cereuisiae fermentum: milk-acid bacteria=1:0.6:0.6,28 ℃ of lower standing for fermentation 36h in constant incubator; With 1:4.05(g/ml) ratio in fermented product, add sterilized water, in the water bath with thermostatic control vibrator, under 30 ℃, with 125r/min rotating speed cyclotron oscillation mode lixiviate 4.5h, after lixiviate finished, with the centrifugal 20min of 4500r/min, supernatant liquor vacuum filtration under vacuum tightness 0.1MPa obtained suction filtration liquid; Selecting molecular weight cut-off is the polysulfone hollow fibre ultra-filtration membrane of 30kDa, 0.075MPa pressure, under 28 ℃ to suction filtration liquid ultra-filtration and separation purifying, get and see through the dehydrated alcohol that liquid adds 4 times of volumes, hold over night in the refrigerator, with the centrifugal 20min of 4500r/min, supernatant liquor is under-55 ℃, vacuum tightness 0.1MPa, and lyophilize 48h obtains non-starch polysaccharide.
The yield of non-starch polysaccharide is 78.37%, purity is 97.28%, the impurity protein content is 0.37%; Antioxidation activity in vitro comprises: the IC that removes hydroxy radical qiao, ultra-oxygen anion free radical, DPPH free radical and ABTS free radical
50Value is respectively 0.84mg/mL, 0.58mg/mL, 0.77mg/mL and 0.38mg/mL; The function of blood sugar reduction activity comprises: the external IC that alpha-glucosidase is suppressed activity
50Value is 54.69mg/L, the hypoglycemic activity of the diabetic mice that streptozotocin, adrenalin hydrochloride and tetraoxypyrimidine are induced, the fasting blood sugar after the peanut non-starch polysaccharide of the feeding treatment experiment have reduced respectively 88.76%, 90.04% and 92.89% before than experiment.
Embodiment 2
Peanut meal was pulverized 120 mesh sieves, and peanut meal powder and the nutrient salt solution got under the sieve (contain 0.55% (NH in the 1L volume
4)
2SO
4, 0.45% KH
2PO
4, 0.9% urea, 1.1% glucose) at 121 ℃ of sterilization 15min, with 1:1.50(g/ml) ratio in the peanut meal powder, add nutrient salt solution, mix, with 1:0.18(g/ml, 10
8Individual/the ml bacterial classification) ratio access cell age in the peanut meal powder be that the aspergillus oryzae of 60h, cereuisiae fermentum and the cell age that cell age is 30h are the mixed strains of the milk-acid bacteria of 9h, the blending ratio of bacterial classification is aspergillus oryzae: cereuisiae fermentum: milk-acid bacteria=1:1.2:0.8,25 ℃ of lower standing for fermentation 30h in constant incubator; With 1:3.85(g/ml) ratio in fermented product, add sterilized water, in the water bath with thermostatic control vibrator, under 25 ℃, with 125r/min rotating speed cyclotron oscillation mode lixiviate 5.0h, after lixiviate finished, with the centrifugal 20min of 4500r/min, supernatant liquor vacuum filtration under vacuum tightness 0.1MPa obtained suction filtration liquid; Selecting molecular weight cut-off is the polysulfone hollow fibre ultra-filtration membrane of 30kDa, 0.075MPa pressure, under 28 ℃ to suction filtration liquid ultra-filtration and separation purifying, get and see through the dehydrated alcohol that liquid adds 4 times of volumes, hold over night in the refrigerator, with the centrifugal 20min of 4500r/min, supernatant liquor is under-55 ℃, vacuum tightness 0.1MPa, and lyophilize 48h obtains non-starch polysaccharide.
The yield of non-starch polysaccharide is 79.26%, purity is 98.05%, the impurity protein content is 0.25%; Antioxidation activity in vitro comprises: the IC that removes hydroxy radical qiao, ultra-oxygen anion free radical, DPPH free radical and ABTS free radical
50Value is respectively 0.76mg/mL, 0.49mg/mL, 0.59mg/mL and 0.31mg/mL; The function of blood sugar reduction activity comprises: the external IC that alpha-glucosidase is suppressed activity
50Value is 50.72mg/L, the hypoglycemic activity of the diabetic mice that streptozotocin, adrenalin hydrochloride and tetraoxypyrimidine are induced, the fasting blood sugar after the peanut non-starch polysaccharide of the feeding treatment experiment have reduced respectively 89.38%, 92.72% and 94.05% before than experiment.
Embodiment 3
Peanut meal was pulverized 120 mesh sieves, and peanut meal powder and the nutrient salt solution got under the sieve (contain 0.55% (NH in the 1L volume
4)
2SO
4, 0.45% KH
2PO
4, 0.9% urea, 1.1% glucose) at 121 ℃ of sterilization 15min, with 1:1.35(g/ml) ratio in the peanut meal powder, add nutrient salt solution, mix, with 1:0.2(g/ml, 10
8Individual/the ml bacterial classification) ratio access cell age in the peanut meal powder be that the aspergillus oryzae of 84h, cereuisiae fermentum and the cell age that cell age is 40h are the mixed strains of the milk-acid bacteria of 15h, the blending ratio of bacterial classification is aspergillus oryzae: cereuisiae fermentum: milk-acid bacteria=1:1.0:1.0,30 ℃ of lower standing for fermentation 40h in constant incubator; With 1:4.25(g/ml) ratio in fermented product, add sterilized water, in the water bath with thermostatic control vibrator, under 35 ℃, with 125r/min rotating speed cyclotron oscillation mode lixiviate 5.5h, after lixiviate finished, with the centrifugal 20min of 4500r/min, supernatant liquor vacuum filtration under vacuum tightness 0.1MPa obtained suction filtration liquid; Selecting molecular weight cut-off is the polysulfone hollow fibre ultra-filtration membrane of 30kDa, 0.075MPa pressure, under 28 ℃ to suction filtration liquid ultra-filtration and separation purifying, get and see through the dehydrated alcohol that liquid adds 4 times of volumes, hold over night in the refrigerator, with the centrifugal 20min of 4500r/min, supernatant liquor is under-55 ℃, vacuum tightness 0.1MPa, and lyophilize 48h obtains non-starch polysaccharide.
The yield of non-starch polysaccharide is 81.57%, purity is 98.36%, the impurity protein content is 0.22%; Antioxidation activity in vitro comprises: the IC that removes hydroxy radical qiao, ultra-oxygen anion free radical, DPPH free radical and ABTS free radical
50Value is respectively 0.71mg/mL, 0.35mg/mL, 0.61mg/mL and 0.26mg/mL; The function of blood sugar reduction activity comprises: the external IC that alpha-glucosidase is suppressed activity
50Value is 48.35mg/L, the hypoglycemic activity of the diabetic mice that streptozotocin, adrenalin hydrochloride and tetraoxypyrimidine are induced, the fasting blood sugar after the peanut non-starch polysaccharide of the feeding treatment experiment have reduced respectively 89.87%, 93.18% and 95.76% before than experiment.
The above is the specific embodiment of the present invention only, is not limited to this, anyly is familiar with those skilled in the art in the technical scope that the present invention discloses, and can expect easily changing or replacing, and all should be encompassed within protection scope of the present invention.
Claims (4)
1. a solid state fermentation prepares the method for peanut non-starch polysaccharide, it is characterized in that, may further comprise the steps:
One, solid state fermentation: peanut meal is pulverized, and crosses 120 mesh sieves, gets the lower peanut meal powder of sieve as raw material, and accesses mixed strains, solid state fermentation in constant incubator after the nutrient salt solution sterilization;
Two, lixiviate: add sterilized water in the fermented product, lixiviate in the water bath with thermostatic control vibrator, centrifugal, vacuum filtration;
Three, separation and purification: suction filtration liquid is got the dehydrated alcohol that adds 4 times of volumes through liquid through the ultra-filtration membrane separation and purification, and hold over night in the refrigerator is centrifugal, and the precipitation lyophilize obtains the peanut non-starch polysaccharide.
2. solid state fermentation according to claim 1 prepares the method for peanut non-starch polysaccharide, it is characterized in that: the nutrient salt solution in the described step 1 is to contain 0.55% (NH in the 1L volume
4)
2SO
4, 0.45% KH
2PO
4, 0.9% urea, 1.1% glucose, the condition of sterilization is 121 ℃ of sterilization 15min, the quality of peanut meal powder and nutrient salt solution and volume ratio are 1:0.5～1.75(g/ml), the bacterial classification of access is that cell age is that the aspergillus oryzae of 60～84h, cereuisiae fermentum and the cell age that cell age is 30～40h are the mixed strains of the milk-acid bacteria of 9～15h, the blending ratio of bacterial classification is aspergillus oryzae: cereuisiae fermentum: milk-acid bacteria=1:0.5～1.5:0.5～1.0, the access volume ratio of peanut meal powder and bacterial classification is 1:0.1～0.2(g/ml, 10
8Individual/the ml bacterial classification), solid state fermentation conditions is 25～30 ℃ of lower standing for fermentation 30～40h.
3. solid state fermentation according to claim 1 prepares the method for peanut non-starch polysaccharide, it is characterized in that: the fermented product in the described step 2 and the quality of sterilized water and volume ratio are 1:3.5～4.5(g/ml), the water bath with thermostatic control extracting condition is under 25～35 ℃, in the water bath with thermostatic control vibrator, with 125r/min rotating speed cyclotron oscillation mode lixiviate 4～6h, centrifugal condition is the centrifugal 20min of 4500r/min, and vacuum filtration obtains suction filtration liquid under the vacuum tightness 0.1MPa.
4. solid state fermentation according to claim 1 prepares the method for peanut non-starch polysaccharide, it is characterized in that: the ultra-filtration membrane purification condition in the described step 3 is that to select molecular weight cut-off be the polysulfone hollow fibre ultra-filtration membrane of 30kDa, 0.075MPa pressure, 28 ℃ of lower ultrafiltration, centrifugal condition is the centrifugal 20min of 4500r/min, the lyophilize condition is-55 ℃, vacuum tightness 0.1MPa, dry 48h.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104161172A (en) * | 2014-06-24 | 2014-11-26 | 山东鲁花集团有限公司 | Method for producing probiotic peptides by using two-step solid-state fermentation of peanut meal |
| CN117843823A (en) * | 2024-01-04 | 2024-04-09 | 广东海天创新技术有限公司 | Method for producing peanut polysaccharide and polyphenol by using cold-pressed peanut meal |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101664167A (en) * | 2009-09-24 | 2010-03-10 | 山东省花生研究所 | Peanut water-soluble dietary fiber enzymatic extracting method |
| CN102121036A (en) * | 2010-11-21 | 2011-07-13 | 山东省花生研究所 | Method for preparing peanut dietary fibers by using microbial fermentation method |
| CN103053787A (en) * | 2013-02-05 | 2013-04-24 | 杨庆利 | Method for preparing peanut peptide by microbial solid fermentation of peanut meal |
-
2013
- 2013-08-07 CN CN201310341028.7A patent/CN103361395B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101664167A (en) * | 2009-09-24 | 2010-03-10 | 山东省花生研究所 | Peanut water-soluble dietary fiber enzymatic extracting method |
| CN102121036A (en) * | 2010-11-21 | 2011-07-13 | 山东省花生研究所 | Method for preparing peanut dietary fibers by using microbial fermentation method |
| CN103053787A (en) * | 2013-02-05 | 2013-04-24 | 杨庆利 | Method for preparing peanut peptide by microbial solid fermentation of peanut meal |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104161172A (en) * | 2014-06-24 | 2014-11-26 | 山东鲁花集团有限公司 | Method for producing probiotic peptides by using two-step solid-state fermentation of peanut meal |
| CN117843823A (en) * | 2024-01-04 | 2024-04-09 | 广东海天创新技术有限公司 | Method for producing peanut polysaccharide and polyphenol by using cold-pressed peanut meal |
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| CN103361395B (en) | 2015-10-14 |
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