CN102860523B - Bone calcium extraction method by double fermentation and bone gla protein - Google Patents
Bone calcium extraction method by double fermentation and bone gla protein Download PDFInfo
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Abstract
The invention discloses a bone calcium extraction method by double fermentation. The bone calcium extraction method by double fermentation includes the steps of fermenting before enzymolysis, enzymolysis and fermenting after enzymolysis. Free calcium content of bone gla protein made by the bone calcium extraction method compared with the prior art is increased greatly. The obtained bone gla protein has fine taste and fragrance, and peculiar odors in extract are removed.
Description
Technical field
The present invention relates to bone calcium extraction process, belong to food processing field.
Background technology
Along with social aging, in western countries, the illness rate of osteoporosis occupies first of metabolic bone disease.The misery that the osteoporosis illness brings patient and family is self-evident.According to statistics, China approximately has the patients with osteoporosis that surpasses 100,000,000 at present, expects the year two thousand fifty will be increased to 200,000,000 1 thousand ten thousand people.Cause the factor of increasing people's calcium deficiency: live and work rhythm is accelerated, operating pressure is large and live irregular, the structure that is not careful in one's diet reasonably combined, calcium was taken in and was less than 600 milligrams of persons and can thinks the calcium insufficiency of intake every day, easily caused the shortage of calcium in body; Some a middle-aged person's work strains, lack self health consciousness, thinks little of outdoor activity, and proper interior synthetic vitamin D is reduced, and affects absorption and the utilization of calcium; Unbalanced or the necessary nutrient of shortage human body is taken in nutrition.
Chinese patent CN1087793A and U.S. Pat 6342252B1 disclose a kind of enzyme solution that utilizes and extracted the technique of calcium element from ox bones.Enzymolysis bone calcium has suitable, nutritious, the calcareous absorption rate high of calcium phosphorus ration.From the market sale situation, enzymolysis bone calcium is subject to liking of consumers in general deeply, takes the osteoporotic successful of rear improvement, has obtained significant economic benefit.
" variation of ultramicro grinding yak bone paste free calcium and amino-acid nitrogen after fermentation and enzymolysis " (Chen Dan, Zhang Chuanlin etc., total the 178th phase of " China brewages " the 1st phase in 2008) put down in writing and taked zymotechnique can increase solution Free Calcium content to bone mud, after lactobacillus-fermented is processed bone mud, free calcium content is 24 times of enzymolysis processing bone mud.
In the bone calcium that above-mentioned technique makes, although by fermentation, free calcium content obviously increases, the mouthfeel of end product is poor, and the taste of the smelling of fish or mutton is difficult to remove.Therefore finding a kind ofly can either significantly increase extract Free Calcium content and keep nutritional labeling in product not run off, and can remove peculiar smell in product again, strengthens the technique of mouthfeel, becomes technical staff's urgent problem.
Summary of the invention
The object of the present invention is to provide a kind of extraction process of new BGP, described technique can better increase free calcium content and keep abundant nutritional labeling than prior art.
Another object of the present invention is to provide a kind of more healthy calcium supplementing product production technology, and the calcium supplementing product that described technique makes has splendid mouthfeel.
Another object of the present invention is to provide a kind of calcium supplementing product with market competitiveness, and described product does not increase production cost because of the design of zymotechnique,
For achieving the above object, the present invention adopts following technical proposal to realize.
The aggregate of the method that the present invention adopts fermentation before to enzymolysis carries out pretreatment, and then carries out enzymolysis and extraction calcium element, and then carries out fermentation process, removes the peculiar smell in BGP, improves mouthfeel.
For strengthening ferment effect, the present invention preferably is broken to bone meal 5～40mm particle, the too small production cost that increases of particle, and the excessive effect of particle is poor.The inventor, through test of many times and experience, finally determines that 5～40mm granulometric range reaches the best cost performance of the extraction of calcium element and cost.
Enzymolysis primary fermentation of the present invention adopts lactic acid bacteria and aggregate by certain weight proportion, and 35～50 ℃, fermentation 12h.The weight proportion of lactic acid bacteria and aggregate, lactic acid bacteria concentration, fermentation temperature and the time in zymotic fluid has important impact to the product Free Calcium content finally made, and the weight proportion of lactic acid bacteria and aggregate and the lactic acid bacteria concentration in zymotic fluid having the greatest impact to free calcium.The inventor is working experience for many years according to oneself, determines after repetition test that it is 0.01～0.04% that lactic acid bacteria accounts for the aggregate weight ratio, under the condition of aggregate and water w/v 1: 3～5, can reach the best cost performance of input and output.
The inventor provides great many of experiments, it is unexpected that the discovery various lactobacillus fermented according to certain proportioning combination, final product Free Calcium content improves more than 8% with certain lactobacillus-fermented more separately, and it is better to adopt bifidobacterium lactis (B.lactis), bulgarian milk bar mattress (L.bulgaricus) and three kinds of lactic acid bacterias of Lactobacillus helveticus (L.helviticus) to carry out ferment effect; Adopt these three kinds of lactic acid bacteria proportioning after fermentation effects best, when three kinds of lactic acid bacterias are combined with weight ratio at 1: 3: 2, after the more single lactobacillus-fermented of enzymolysis liquid Free Calcium content, improve 10%～30%.
Secondary fermentation of the present invention adopts leuconostoc cremoris and citric acid, and the proportioning between the two all has important impact to taste, the production efficiency of end product.The inventor finally determines that under the condition of leuconostoc cremoris and citric acid weight ratio 1: 20～30, the end product effect of fermentation is better.Under these conditions, determine that fermentation temperature is 39 ℃, when fermentation time is 12h, can reach the optimization of production efficiency.
The present invention adopts the method for high-temperature sterilization to reach the purpose of sterilizing, also removes the lactic acid bacteria in enzymolysis liquid simultaneously.
Enzymolysis of the present invention adopts the secondary enzymolysis method, and after screening, final discovery is carried out enzymolysis with pepsin and papain and had better effect than other protease.Certainly, select other protease to carry out enzymolysis and also can realize purpose of the present invention.Enzymolysis scheme best in extraction process of the present invention is pH value 1～3, adds pepsin, and 35～40 ℃ of hydrolysis temperatures, stir 1～2h; PH value 6～7, add papain, 40～60 ℃, stirs 1～2h.
It is 0.04～0.08% for good that hydrolysis bone meal two kinds of protease used respectively account for the aggregate weight ratio.
The various formulations that on composition of the present invention and any or more than one pharmacies, auxiliary material is mixed as starch, dextrin, lactose, microcrystalline cellulose, HPMC, polyethylene glycol, dolomol, superfine silica gel powder, xylitol, lactitol, glucose, glycine, sweet mellow wine, glycine etc., for example, can be made into tablet, sustained release tablets, dripping pill, granule, capsule, pulvis, fine granule.Preferred dosage form is tablet or granule.
The BGP that technique of the present invention makes than prior art, free calcium content improves 8%～30%.The product mouthfeel finally made is good, product fragrance, and the peculiar smell in extract is removed.
Test example
Below by some, concrete experimental data further illustrates beneficial effect of the present invention, all embodiment of the present invention all can make close therewith experiment effect, following data just illustrate.
1, materials and methods
1.1 ox bone: be purchased from the Hebei good fortune and become five rich food limited companies
1.2 sample 1: the sample that uses ox bone to obtain according to Chinese patent CN1087793A; Sample 2: the sample that uses ox bone to obtain according to U.S. Pat 6342252B1; Sample 3: according to " variation of ultramicro grinding yak bone paste free calcium and amino-acid nitrogen after fermentation and enzymolysis " (Chen Dan, Zhang Chuanlin etc., total the 178th phase of " China brewages " the 1st phase in 2008) in, " 1.5 lactobacillus-fermenteds are processed yak bone pastes ... the concentration that ox bone mud is 10% with distilled water diluting ... ferment 36 hours " used the sample of ox bone acquisition; Sample 4: the sample that uses ox bone to obtain according to the embodiment of the present invention 3.
1.3 method: adopt atomic absorption spectroscopy determination free calcium content.
2, result
2.1 the free calcium content of sample 1 is 192.19mg/100g, the free calcium content of sample 2 is 341.07mg/100g, and the free calcium content of sample 3 is 2076.82mg/100g, and the free calcium content of sample 4 is 2528.54mg/100g.
2.2, through the Data Comparison analysis, the free calcium content of the prepared product of the present invention, compare with sample 1, sample 2, has the raising of highly significant; Compare with sample 3, have significant raising, increase rate reaches 21.75%.
The specific embodiment
Embodiment 1
Preparation culture medium: MRS culture medium: peptone 1%, beef extract 1%, yeast extract 0.5%, glucose 2%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, magnesium sulfate 0.02%, manganese sulfate 0.005%, Tween-80 0.1%, Triammonium citrate 0.2%, PH5.5～6.0,115 ℃, sterilizing 30min.
Embodiment 2
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, and 40 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.02% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 3
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 3, it is 0.01% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 35 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 1, add pepsin, accounting for the aggregate weight ratio is 0.04%, and 35 ℃ of hydrolysis temperatures stir 1h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.04%, 40 ℃, stirs 1h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.06% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 20, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 4
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 4, it is 0.03% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 40 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 2, add pepsin, accounting for the aggregate weight ratio is 0.06%, and 37 ℃ of hydrolysis temperatures stir 1.5h, adds alkali and regulates pH value to 6.5, adds papain, and accounting for the aggregate weight ratio is 0.06%, 50 ℃, stirs 1.5h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.04% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 25, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 5
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 1, add pepsin, accounting for the aggregate weight ratio is 0.08%, and 37 ℃ of hydrolysis temperatures stir 1h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.06%, 45 ℃, stirs 1h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.03% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 6
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 3, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 40 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, and 40 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 1h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 25, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 7
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 45 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 1, add pepsin, accounting for the aggregate weight ratio is 0.07%, and 39 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.07%, 55 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 22, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 8
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 4, it is 0.03% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 2.5, add pepsin, accounting for the aggregate weight ratio is 0.04%, and 36 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 6.5, adds papain, and accounting for the aggregate weight ratio is 0.08%, 58 ℃, stirs 1.5h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.04% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 9
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 3.5, it is 0.01% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 35 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 1.5, add pepsin, accounting for the aggregate weight ratio is 0.08%, and 38 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.08%, 40 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.03% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 10
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 3, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.06%, and 40 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.07%, 50 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 28, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 11
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 4.5, it is 0.02% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 40 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 2, add pepsin, accounting for the aggregate weight ratio is 0.05%, and 38 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.07%, 55 ℃, stirs 1h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.02% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 26, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 12
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 42 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 2, add pepsin, accounting for the aggregate weight ratio is 0.06%, and 39 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.04%, 55 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 20, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 13
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 4, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 45 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, and 37 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.08%, 52 ℃, stirs 1.5h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 14
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 3.5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 2, add pepsin, accounting for the aggregate weight ratio is 0.06%, and 40 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.06% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 15
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 1.5, add pepsin, accounting for the aggregate weight ratio is 0.06%, and 37 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 50 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.03% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 26, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 16
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 4, it is 0.02% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 40 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, and 40 ℃ of hydrolysis temperatures stir 1h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 45 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 17
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 45 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.06%, and 36 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.04%, 55 ℃, stirs 1.5h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.03% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 20, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 18
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 5, it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 3, add pepsin, accounting for the aggregate weight ratio is 0.04%, and 40 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 2h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.05% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 25, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Embodiment 19
Bone is pulverized, sterilizing; Get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Aggregate and water w/v 1: 4.5, it is 0.03% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 48 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 2, add pepsin, accounting for the aggregate weight ratio is 0.04%, and 40 ℃ of hydrolysis temperatures stir 2h, adds alkali and regulates pH value to 6, adds papain, and accounting for the aggregate weight ratio is 0.06%, 50 ℃, stirs 1h, heats the enzyme that goes out; Add 5% glucose of gross weight, add leuconostoc cremoris and citric acid, it is 0.03% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1: 20, fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
Claims (2)
1. the method for a secondary fermentation extraction bone calcium, comprise the steps:
(1) bone is pulverized, lactobacillus-fermented, heat sterilization;
(2) enzymolysis;
(3) lactobacillus-fermented, heat sterilization, be drying to obtain;
It is 0.01～0.04% that the described lactic acid bacteria of step (1) accounts for the aggregate weight ratio, aggregate and water w/v 1:3～5;
Fermentation is 35～50 ℃, fermentation 12h;
The described lactic acid bacteria of step (1) is bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L. bulgaricus) and Lactobacillus helveticus (L. helviticus);
The weight ratio of described three kinds of lactic acid bacterias is 1:3:2;
The described lactic acid bacteria of step (3) is leuconostoc cremoris, adds citric acid in zymotic fluid;
It is 0.02～0.06% that described leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1:20～30;
Fermentation temperature is 39 ℃, and fermentation time is 12h;
The described enzymolysis scheme of step (2) is pH value 1～3, adds pepsin, and 35～40 ℃ of hydrolysis temperatures, stir 1～2h; PH value 6～7, add papain, 40～60 ℃, stirs 1～2h;
It is 0.04～0.08% that described hydrolysis aggregate two kinds of protease used respectively account for the aggregate weight ratio.
2. a BGP that is extracted preparation by animal skeleton, is characterized in that, adopts the following methods preparation: bone is pulverized, sterilizing; Get bifidobacterium lactis (B.1actis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratio 1:3:2; Aggregate and water w/v 1:3～5, it is 0.01～0.04% that lactic acid bacteria accounts for the aggregate weight ratio, adds 5% sucrose of gross weight, 35～50 ℃, fermentation 12h, 120 ℃ of sterilizing 30min; Add hydrochloric acid adjust pH 1～3, add pepsin, accounting for the aggregate weight ratio is 0.04～0.08%, 35～40 ℃ of hydrolysis temperatures, stir 1～2h, add alkali and regulate pH value to 6～7, add papain, accounting for the aggregate weight ratio is 0.04～0.08%, 40～60 ℃, stir 1～2h, heat the enzyme that goes out; 5% the glucose that adds gross weight, add leuconostoc cremoris and citric acid, it is 0.02～0.06% that leuconostoc cremoris accounts for the aggregate weight ratio, leuconostoc cremoris and citric acid weight ratio 1:20～30, fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, 120 ℃ of sterilizings, be drying to obtain.
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| CN103340413B (en) * | 2013-06-09 | 2014-12-03 | 哈尔滨天鹅土畜产技术开发公司 | Preparation method for bone calcium powder from outer wall of bone through lactobacillus fermentation |
| CN107751815A (en) * | 2017-09-18 | 2018-03-06 | 天津天狮生物发展有限公司 | A kind of production method for digesting bone calcium powder |
| CN108850421A (en) * | 2018-05-04 | 2018-11-23 | 天津天狮生物发展有限公司 | A kind of method and osteocalcin for extracting bone calcium |
| CN108741093A (en) * | 2018-05-24 | 2018-11-06 | 广西山水牛畜牧业有限责任公司 | The extracting method of calcium in a kind of ox bone |
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