CN102871123A - Method for preparing bone calcium and bone gla protein - Google Patents
Method for preparing bone calcium and bone gla protein Download PDFInfo
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Abstract
The invention discloses a method for preparing bone calcium and bone gla protein prepared by means of the method. The method comprises the following steps of three-time defatting, two-time centrifugation, two-time fermentation and secondary protease hydrolysis. Fat in bone meal prepared by adopting a defatting process is completely removed basically, so that the fat content in calcium obtained by means of enzymolysis is lower than 0.1%. The bone gla protein is safe and healthy to consumers with cardiovascular disease, and economical burden to the consumers is not increased.
Description
Technical field
The present invention relates to bone calcium extraction process, belong to food processing field.
Background technology
Along with the aging of society, 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 has approximately at present and surpasses 100,000,000 patients with osteoporosis, 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 the body; Some a middle-aged person's work strains lack self health consciousness, think little of outdoor activity, and proper interior synthetic vitamin D is reduced, and affect absorption and the utilization of calcium; Nutrition is taken in unbalanced or is lacked the necessary nutrient of human body.
The calcium agent of existing market mostly is the synthetic calcium of industry.The calcium agent nutritional labeling that industry is synthesized is single, is difficult to satisfy many-sided nutritional need in the bone growth metabolic process, and effect of supplemented calcium is undesirable.Animal Bone is rich in the several kinds of mineral elements such as calcium, phosphorus, iron, magnesium, and with people's bone photo seemingly, the skeleton cell has stronger affinity to homologue's cell, thereby utilization rate is higher.
Chinese patent CN1087793A and US Patent No. 6342252B1 disclose a kind of technique of utilizing enzyme solution to extract the calcium element from ox bone.Enzymolysis bone calcium has suitable, nutritious, the calcareous absorption rate high of calcium phosphorus ration.From the market sale situation, enzymolysis bone calcium is subjected to liking of consumers in general deeply, takes the osteoporotic successful of rear improvement, has obtained significant economic benefit.
But in the bone calcium that above-mentioned technique makes, degreasing method mainly is to implement in the boiling mode, is difficult to remove the fat of high-load in the sclerotin, causes final product fat content higher.For the consumer with angiocardiopathy, the hazards that too high fat intake obviously can raise and fall ill.Therefore, from animal skeleton, extract in the process of calcium element, remove excess fat, reduce the onset risk of angiocardiopathy, become 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 enzymolysis bone calcium, described technique can better be removed fat in the sclerotin than prior art.
Another object of the present invention provides a kind of more healthy calcium supplementing product production technology, and the calcium supplementing product that described technique makes has lower fat content.
Another object of the present invention provides a kind of calcium supplementing product with market competitiveness, and described product does not increase production cost because of the design of degreasing process,
For achieving the above object, the present invention adopts following technical proposal to realize.
The present invention adopts three boiling modes to remove animal oil, is 60～80 ℃ for the first time, 45～75min; 90～100 ℃ for the second time, 10min; Be for the third time behind enzymolysis, adopt the mode of high temperature to remove final fat that may be residual.Three boilings also have the effect of sterilization simultaneously except having the fatty effect of removing in the sclerotin.For strengthening degreasing effect, the present invention preferably is broken to bone meal 5～40mm particle, the too small production cost that then increases of particle, and the excessive degrease effect of particle is relatively poor.The inventor determines finally that through test of many times and experience 5～40mm granulometric range reaches the best cost performance of bone fat and cost.
After boiling, the present invention also adopts backwashing manner to remove the fat that exists in the aggregate; Again with 4000～8000r/min, centrifugal 30min can remove most fat in the aggregate like this after the flushing.
For further removing remaining fat, process choice of the present invention again adds alkaline lipase and carries out fat remaining in the enzymolysis aggregate after centrifugal.Alkaline lipase A, B account for 0.1～0.4%, 20～30 ℃ of aggregate weight ratios by 1.5～3: 1 compound lipases that mixes, hydrolysis 40～60min.Behind enzymolysis, secondary centrifuging is further removed remaining trace fat.
The present invention adopts the method for fermentation that the aggregate before fermenting is carried out preliminary treatment, and then carries out enzymolysis and extraction calcium element, and then extract is carried out fermentation process, removes the peculiar smell in the extract, improves mouthfeel.
For strengthening ferment effect, the present invention preferably is broken to bone meal 5～40mm particle, the too small production cost that then increases of particle, and the excessive effect of particle is relatively poor.The inventor determines finally that through test of many times and experience 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 that finally makes, 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 determines after the repetition test that according to oneself working experience for many years 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 various lactobacillus of finding ferment 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), lactobacillus bulgaricus (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 make up with weight ratio at 1: 3: 2, improve 10%～30% behind the more single lactobacillus-fermented of enzymolysis liquid Free Calcium content.
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 determines that finally the end product effect of fermentation is better under the condition of leuconostoc cremoris and citric acid weight ratio 1: 20～30.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 the 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 the extraction process of the present invention is pH value 1～3, adds pepsin, 35～40 ℃ of hydrolysis temperatures, 1～2h; Transfer pH value 6～7, add papain, 40～60 ℃, stir 1～2h.
It is 0.04～0.08% for good that hydrolysis two kinds of used protease of aggregate respectively account for the aggregate weight ratio.
The alkaline lipase that alkaline lipase A is produced by the Penicillium expansion PF868 of bioengineering institute of Fujian Normal University development, Co., Ltd provides by the green little health bioengineering in Shenzhen; Alkaline lipase B adopts biotechnology to be made with extra care and the alkaline lipase of generation by fungi fermentation, is provided by Haining Jin Chao Industrial Co., Ltd..
The various formulations that auxiliary material such as starch, dextrin, lactose, microcrystalline cellulose, HPMC, polyethylene glycol, dolomol, superfine silica gel powder, xylitol, lactitol, glucose, glycine, sweet mellow wine, glycine etc. are mixed on composition of the present invention and any or more than one pharmacies, for example, can be made into tablet, sustained release tablets, dripping pill, granule, capsule, fine granule.Preferred dosage form is tablet or granule.
Adopt in the bone meal that technique of the present invention makes, fat is completely removed substantially, so that fat content is lower than 0.1% in the calcium agent that enzymolysis obtains.Product safety and health more concerning the consumer that angiocardiopathy is arranged, but financial burden therefore increased again.
Test example
Below further specify beneficial effect of the present invention by some concrete experimental datas, 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 the ox bone acquisition according to Chinese patent CN1087793A; Sample 2: the sample that uses the ox bone acquisition according to US Patent No. 6342252B1; Sample 3: according to " variation of ultramicro grinding yak bone paste free calcium and amino-acid nitrogen behind fermentation and enzymolysis " (Chen Dan, Zhang Chuanlin etc., total the 178th phase of " China brewages " the 1st phase in 2008) sample of " 1.5 lactobacillus-fermenteds are processed yak bone paste ... ox bone mud distilled water diluting is 10% concentration ... ferment 36 hours " use ox bone acquisition in; Sample 4: the sample that uses the ox bone acquisition according to the embodiment of the invention 4.
1.3 method: adopt atomic absorption spectroscopy determination free calcium content.
2, result
2.1 the free calcium content of sample 1 is 170.80mg/100g, the free calcium content of sample 2 is 343.13mg/100g, and the free calcium content of sample 3 is 1997.01mg/100g, and the free calcium content of sample 4 is 2387.45mg/100g.
2.2 through the Data Comparison analysis, the free calcium content of the prepared product of the present invention is compared with sample 1, sample 2, has the raising of highly significant; Compare with sample 3, have significant raising, increase rate reaches 19.55%.
The specific embodiment
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 ℃, sterilization 30min.
Embodiment 1
Get animal skeleton and be crushed to the 40mm particle, add ethanol, 80 ℃, lixiviate 75min abandons supernatant; 90 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 10, add alkaline lipase A, B by the compound lipases of mixing in 1.5: 1, account for 0.4%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets 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 add 5% sucrose of gross weight, and it is 0.04%, 50 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, 40 ℃ of hydrolysis temperatures, 2h; Heating is desalted acid to pH value 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 2h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.01%.
Embodiment 2
Get animal skeleton and be crushed to the 5mm particle, add ethanol, 60 ℃, lixiviate 45min abandons supernatant; 80 ℃ of hot-water soak rinsing 30min, with aggregate 4000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 3 are transferred pH value 8, add alkaline lipase A, B by the compound lipases of mixing in 3: 1, account for 0.1%, 20 ℃ of aggregate weight ratio, and hydrolysis 40min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets 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 add 5% sucrose of gross weight, and it is 0.01%, 35 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 1, add pepsin, accounting for the aggregate weight ratio is 0.04%, 35 ℃ of hydrolysis temperatures, 1h; Heating is desalted acid to pH value 6, adds papain, and accounting for the aggregate weight ratio is 0.04%, 40 ℃, stirs 1h, separating liquid; 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: 20, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.02%.
Embodiment 3
Get animal skeleton and be crushed to the 20mm particle, add ethanol, 70 ℃, lixiviate 60min abandons supernatant; 85 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 4 are transferred pH value 9, add alkaline lipase A, B by the compound lipases of mixing in 2: 1, account for 0.3%, 25 ℃ of aggregate weight ratio, and hydrolysis 50min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets 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 add 5% sucrose of gross weight, and it is 0.03%, 40 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 2, add pepsin, accounting for the aggregate weight ratio is 0.06%, 37 ℃ of hydrolysis temperatures, 1.5h; Heating is desalted acid to pH value 6.5, adds papain, and accounting for the aggregate weight ratio is 0.06%, 50 ℃, stirs 1.5h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.02%.
Embodiment 4
Get animal skeleton and be crushed to the 10mm particle, add ethanol, 80 ℃, lixiviate 75min abandons supernatant; 90 ℃ of hot-water soak rinsing 30min, with aggregate 6000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 3.5 are transferred pH value 8.5, add alkaline lipase A, B by the compound lipases of mixing in 2: 1, account for 0.2%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacterias; Aggregate and water w/v 1: 5 add 5% sucrose of gross weight, and it is 0.04%, 50 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 1, add pepsin, accounting for the aggregate weight ratio is 0.08%, 37 ℃ of hydrolysis temperatures, 1～2h; Heating is desalted acid to pH value 6～7, adds papain, and accounting for the aggregate weight ratio is 0.06%, 45 ℃, stirs 1h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.03%.
Embodiment 5
Get animal skeleton and be crushed to the 25mm particle, add ethanol, 75 ℃, lixiviate 65min abandons supernatant; 86 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 4 are transferred pH value 9.5, add alkaline lipase A, B by the compound lipases of mixing in 2.8: 1, account for 0.3%, 30 ℃ of aggregate weight ratio, and hydrolysis 50min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2; Add 5% sucrose of gross weight, it is 0.04%, 40 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Transfer pH value 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, 40 ℃ of hydrolysis temperatures, 2h; Heating is desalted acid to pH value 7, adds papain, and accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 1h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.01%.
Embodiment 6
Get animal skeleton and be crushed to the 35mm particle, add ethanol, 78 ℃, lixiviate 70min abandons supernatant; 87 ℃ of hot-water soak rinsing 30min, with aggregate 7000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 8, add alkaline lipase A, B by the compound lipases of mixing in 1.8: 1, account for 0.25%, 20 ℃ of aggregate weight ratio, and hydrolysis 50min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacterias; Aggregate and water w/v 1: 5 add 5% sucrose of gross weight, and it is 0.04%, 45 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 1, add pepsin, 35～40 ℃ of hydrolysis temperatures, 1～2h; Heating is desalted acid to pH value 7, adds papain, 55 ℃, stirs 2h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.02%.
Embodiment 7
Get animal skeleton and be crushed to the 10mm particle, add ethanol, 60 ℃, lixiviate 75min abandons supernatant; 88 ℃ of hot-water soak rinsing 30min, with aggregate 5000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 4 are transferred pH value 10, add alkaline lipase A, B by the compound lipases of mixing in 3: 1, account for 0.35%, 24 ℃ of aggregate weight ratio, and hydrolysis 45min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacterias add 5% sucrose of gross weights, it is 0.03% that lactic acid bacteria accounts for the aggregate weight ratio, 50 ℃, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 1～3, add pepsin, 35～40 ℃ of hydrolysis temperatures, 2h, heating is desalted acid to pH value 7, adds papain, 40～60 ℃, stirs 2h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.02%.
Embodiment 8
Get animal skeleton and be crushed to the 15mm particle, add ethanol, 80 ℃, lixiviate 75min abandons supernatant; 89 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 8.5, add alkaline lipase A, B by the compound lipases of mixing in 2.6: 1, account for 0.14%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios 1: 3: 2,5% the sucrose that adds gross weight, 35～50 ℃, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 1～3, add pepsin, 35～40 ℃ of hydrolysis temperatures, 1～2h; Heating is desalted acid to pH value 6, adds papain, 40 ℃, stirs 2h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.03%.
Embodiment 9
Get animal skeleton and be crushed to the 30mm particle, add ethanol, 75 ℃, lixiviate 55min abandons supernatant; 85 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 8, add alkaline lipase A, B by the compound lipases of mixing in 3: 1, account for 0.35%, 28 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, and the extracting lactic acid bacterium joins in the aggregate, and it is 0.04% that lactic acid bacteria accounts for the aggregate weight ratio, and aggregate and water w/v 1: 3 add 5% sucrose of gross weight, and 50 ℃, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 3, add pepsin, 35～40 ℃ of hydrolysis temperatures, 2h; Heating is desalted acid to pH value 7, adds papain, 50 ℃, stirs 2h, separating liquid; 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: 28, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.02%.
Embodiment 10
Get animal skeleton and be crushed to the 20mm particle, add ethanol, 80 ℃, lixiviate 60min abandons supernatant; 84 ℃ of hot-water soak rinsing 30min, with aggregate 6000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 10, add alkaline lipase A, B by the compound lipases of mixing in 3: 1, account for 0.3%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, get bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus), three kinds of lactic acid bacteria weight ratios add in the aggregate at 1: 3: 2,5% the sucrose that adds gross weight, 40 ℃, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value 2, add pepsin, accounting for the aggregate weight ratio is 0.05%, 38 ℃ of hydrolysis temperatures, 2h; Heating is desalted acid to pH value 6, adds papain, and accounting for the aggregate weight ratio is 0.07%, 55 ℃, stirs 1h, separating liquid; 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: 26, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.03%.
Embodiment 11
Bone is crushed to the 40mm particle, adds ethanol, 80 ℃, lixiviate 75min abandons supernatant; 83 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 10, add the compound lipases that alkaline lipase A, B mix, and account for 0.1%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, and the extracting lactic acid bacterium adds in the aggregate, and aggregate and water w/v 1: 5 add 5% sucrose of gross weight, and 42 ℃, fermentation 12h, 120 ℃ of 30min that sterilize; Transfer pH value, add protease, accounting for the aggregate weight ratio is 0.08%, hydrolysis 2h; Transfer pH value, add protease again, accounting for the aggregate weight ratio is 0.06%, 55 ℃, stirs 2h, separating liquid; 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: 20, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.02%.
Embodiment 12
Bone is crushed to the 30mm particle, adds ethanol, 75 ℃, lixiviate 60min abandons supernatant; 81 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 4 are transferred pH value 9, add compound lipases, account for 0.4%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets 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 add 5% sucrose of gross weight, and it is 0.04%, 50 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value, add protease, accounting for the aggregate weight ratio is 0.06%, 40 ℃ of hydrolysis temperatures, 2h; Transfer pH value, again add protease, accounting for the aggregate weight ratio is 0.08%, 60 ℃, stirs 2h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.03%.
Embodiment 13
Bone is crushed to the 10mm particle, adds ethanol, 80 ℃, lixiviate 75min abandons supernatant; 84 ℃ of hot-water soak rinsing 30min, with aggregate 8000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 5 are transferred pH value 10, add the compound lipases that alkaline lipase A, B mix, and account for 0.4%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, gets 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 add 5% sucrose of gross weight, and it is 0.04%, 50 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min; Add hydrochloric acid and transfer pH value, add pepsin, 37 ℃ of hydrolysis temperatures, 2h; Heating is desalted acid to pH value 6～7, adds neutral proteinase, and accounting for the aggregate weight ratio is 0.08%, 50 ℃, stirs 2h, separating liquid; 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: 26, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.01%.
Embodiment 14
Bone is crushed to the 25mm particle, adds ethanol, 80 ℃, lixiviate 65min abandons supernatant; 84 ℃ of hot-water soak rinsing 30min, with aggregate 6000r/min, centrifugal 30min abandons supernatant; Aggregate after centrifugal is added water, and aggregate and water w/v 1: 4 are transferred pH value 8.5, add compound lipases, account for 0.3%, 30 ℃ of aggregate weight ratio, and hydrolysis 60min is centrifugal, removes supernatant; Centrifugal rear aggregate adds water, and the extracting lactic acid bacterium adds in the aggregate, and it is 0.02% that lactic acid bacteria accounts for the aggregate weight ratio, and aggregate and water w/v 1: 4 add 5% sucrose of gross weight, and 50 ℃, fermentation 12h, 120 ℃ of 30min that sterilize; Add hydrochloric acid and transfer pH value 3, add pepsin, accounting for the aggregate weight ratio is 0.08%, 40 ℃ of hydrolysis temperatures, 1h; Heating is desalted acid to pH value 7, adds neutral proteinase, and accounting for the aggregate weight ratio is 0.08%, 45 ℃, stirs 2h, separating liquid; 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, and fermentation temperature is 39 ℃, and fermentation time is 12h, adds the alkali neutralization, and 120 ℃ of deactivations are drying to obtain.Adopt " Determination of in the GB/T5009.6 food " regulation detection method that bone meal is carried out the determination of fat, recording the product fat content is 0.03%.
Claims (10)
1. the preparation method of a bone calcium comprises the steps:
(1) bone is pulverized, the lixiviate oil of boning is abandoned supernatant;
(2) aggregate after the lixiviate is soaked rinsing, centrifugal, abandon supernatant;
(3) aggregate after centrifugal adds water, transfers pH value, adds the alkaline fat enzyme hydrolysis, and is centrifugal, abandons supernatant;
(4) aggregate after centrifugal in the step (3) is added water, add lactobacillus-fermented, high-temperature inactivation;
(5) zymotic fluid in the step (4) adds the hydrolysis of protease secondary again;
(6) add again lactobacillus-fermented in the hydrolyzate in the step (5), high-temperature inactivation, and get final product.
2. bone calcium preparation method according to claim 1 is characterized in that, described step (1) particles of aggregates 5～40mm adds ethanol, 60～80 ℃, 45～75min.
3. bone calcium preparation method according to claim 1 is characterized in that, in the described step (2) with the rinsing of particles of aggregates hot water, 4000～8000r/min then, centrifugal 30min.
4. bone calcium preparation method according to claim 1 is characterized in that, aggregate and water w/v 1: 3～5 in the described step (3) are transferred pH value 8～10, add the compound lipases that alkaline lipase A, B mix.
5. bone calcium preparation method according to claim 4 is characterized in that, described compound lipases neutral and alkali lipase A, B weight ratio are 1.5～3: 1.
6. according to claim 4 or 5 described bone calcium preparation methods, it is characterized in that described compound lipases accounts for aggregate weight ratio 0.1～0.4%.
7. bone calcium preparation method according to claim 1, it is characterized in that, the described fermentation of step (4) is 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 add 5% sucrose of gross weight, and it is 0.01～0.04%, 35～50 ℃ that lactic acid bacteria accounts for the aggregate weight ratio, fermentation 12h, 120 ℃ of sterilization 30min.
8. bone calcium preparation method according to claim 7 is characterized in that, the described enzymolysis of step (5) is to transfer pH value 1～3, adds pepsin, 35～40 ℃ of hydrolysis temperatures, 1～2h; Transfer pH value 6～7, add papain, 40～60 ℃, stir 1～2h.
9. bone calcium preparation method according to claim 8 is characterized in that, it is 0.04～0.08% that two kinds of used protease of described hydrolysis aggregate respectively account for the aggregate weight ratio.
10. bone calcium preparation method according to claim 1, it is characterized in that, the described fermentation of step (6) is to 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, and fermentation temperature is 39 ℃, fermentation time is 12h, add the alkali neutralization, 120 ℃ of sterilization 30min are drying to obtain.
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| CN103211836A (en) * | 2013-04-11 | 2013-07-24 | 天津天狮生物发展有限公司 | Composition for treating osteoporosis |
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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 |
| CN105077000A (en) * | 2015-07-10 | 2015-11-25 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for preparing fine dried noodles with pork bone soup |
| CN105077000B (en) * | 2015-07-10 | 2018-08-24 | 湖北省农业科学院农产品加工与核农技术研究所 | A kind of preparation method of pig bone made soup vermicelli |
| CN107048218A (en) * | 2017-03-10 | 2017-08-18 | 广西乐业康辉生态养殖专业合作社 | A kind of processing method of bovine bone powder |
| CN108157976A (en) * | 2017-11-30 | 2018-06-15 | 广州城市职业学院 | A kind of two-step fermentation prepares the method for sturgeon activated calcium powder easy to digest and manufactured calcium powder |
| CN108157976B (en) * | 2017-11-30 | 2021-05-04 | 广州城市职业学院 | Method for preparing digestible sturgeon active calcium powder through two-step fermentation and prepared calcium powder |
| CN108741092A (en) * | 2018-05-04 | 2018-11-06 | 天津天狮生物发展有限公司 | A kind of method and osteocalcin of fermentation extraction bone calcium |
| 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 |
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