CN108850421A - A kind of method and osteocalcin for extracting bone calcium - Google Patents
A kind of method and osteocalcin for extracting bone calcium Download PDFInfo
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- CN108850421A CN108850421A CN201810418980.5A CN201810418980A CN108850421A CN 108850421 A CN108850421 A CN 108850421A CN 201810418980 A CN201810418980 A CN 201810418980A CN 108850421 A CN108850421 A CN 108850421A
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- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 52
- 102000004067 Osteocalcin Human genes 0.000 title claims abstract description 17
- 108090000573 Osteocalcin Proteins 0.000 title claims abstract description 17
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 title abstract description 41
- 239000011575 calcium Substances 0.000 title abstract description 41
- 229910052791 calcium Inorganic materials 0.000 title abstract description 41
- 238000000034 method Methods 0.000 title abstract description 19
- 238000000855 fermentation Methods 0.000 claims abstract description 40
- 230000004151 fermentation Effects 0.000 claims abstract description 39
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 78
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 70
- 235000017632 Leuconostoc cremoris Nutrition 0.000 claims description 39
- 241000894006 Bacteria Species 0.000 claims description 35
- 239000004310 lactic acid Substances 0.000 claims description 35
- 235000014655 lactic acid Nutrition 0.000 claims description 35
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 claims description 25
- 230000001954 sterilising effect Effects 0.000 claims description 24
- 241000186018 Bifidobacterium adolescentis Species 0.000 claims description 23
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 22
- 239000003513 alkali Substances 0.000 claims description 22
- 238000002360 preparation method Methods 0.000 claims description 19
- 239000004365 Protease Substances 0.000 claims description 18
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- 102000057297 Pepsin A Human genes 0.000 claims description 17
- 108090000284 Pepsin A Proteins 0.000 claims description 17
- 230000007062 hydrolysis Effects 0.000 claims description 17
- 238000006460 hydrolysis reaction Methods 0.000 claims description 17
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- 235000019834 papain Nutrition 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 13
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
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- 229940009289 bifidobacterium lactis Drugs 0.000 claims description 12
- 102000004190 Enzymes Human genes 0.000 claims description 11
- 108090000790 Enzymes Proteins 0.000 claims description 11
- 229930006000 Sucrose Natural products 0.000 claims description 11
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 11
- 238000001035 drying Methods 0.000 claims description 11
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- 238000006386 neutralization reaction Methods 0.000 claims description 11
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- 230000009849 deactivation Effects 0.000 claims description 10
- 230000037182 bone density Effects 0.000 claims description 8
- 241001465754 Metazoa Species 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 241001468193 Leuconostoc mesenteroides subsp. cremoris Species 0.000 claims 4
- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 claims 2
- 229940079593 drug Drugs 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- 235000013402 health food Nutrition 0.000 claims 1
- 239000000047 product Substances 0.000 abstract description 20
- 230000007071 enzymatic hydrolysis Effects 0.000 abstract description 6
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 abstract description 6
- 239000012467 final product Substances 0.000 abstract description 4
- 239000000284 extract Substances 0.000 abstract description 3
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- 244000172809 Leuconostoc cremoris Species 0.000 description 35
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 24
- 230000000694 effects Effects 0.000 description 22
- 238000012360 testing method Methods 0.000 description 18
- 238000003756 stirring Methods 0.000 description 12
- 239000007788 liquid Substances 0.000 description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 208000001132 Osteoporosis Diseases 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229940036811 bone meal Drugs 0.000 description 2
- 239000002374 bone meal Substances 0.000 description 2
- 238000010959 commercial synthesis reaction Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
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- 230000001502 supplementing effect Effects 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 206010065687 Bone loss Diseases 0.000 description 1
- 206010006956 Calcium deficiency Diseases 0.000 description 1
- 241001269238 Data Species 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 240000002605 Lactobacillus helveticus Species 0.000 description 1
- 235000013967 Lactobacillus helveticus Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 208000030136 Marchiafava-Bignami Disease Diseases 0.000 description 1
- 208000029725 Metabolic bone disease Diseases 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
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- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241001552596 Scorzoneroides helvetica Species 0.000 description 1
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- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930003316 Vitamin D Natural products 0.000 description 1
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000001479 atomic absorption spectroscopy Methods 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000008468 bone growth Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 1
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- 229960001031 glucose Drugs 0.000 description 1
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- 229960002449 glycine Drugs 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 229960003943 hypromellose Drugs 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000000832 lactitol Substances 0.000 description 1
- VQHSOMBJVWLPSR-JVCRWLNRSA-N lactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-JVCRWLNRSA-N 0.000 description 1
- 235000010448 lactitol Nutrition 0.000 description 1
- 229960003451 lactitol Drugs 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 229940054346 lactobacillus helveticus Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000006180 nutrition needs Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- -1 phosphoric acid hydrogen Chemical class 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000033764 rhythmic process Effects 0.000 description 1
- 108010048734 sclerotin Proteins 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
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- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000007939 sustained release tablet Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
- 235000019166 vitamin D Nutrition 0.000 description 1
- 239000011710 vitamin D Substances 0.000 description 1
- 150000003710 vitamin D derivatives Chemical class 0.000 description 1
- 229940046008 vitamin d Drugs 0.000 description 1
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- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/10—Anti-acne agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/513—Adolescentes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/531—Lactis
Abstract
The invention discloses a kind of methods that secondary fermentation extracts bone calcium, include the following steps：Bone crushes, digests primary fermentation, enzymatic hydrolysis and enzymatic hydrolysis post-fermentation.Technique of the present invention is greatly improved compared with osteocalcin made from the prior art, free calcium levels.Final product obtained is in good taste, product fragrance, and the peculiar smell in extract is removed.
Description
Technical field
The present invention relates to bone calcium extraction processes, belong to food processing field.
Background technique
With the aging of society, in western countries, the illness rate of osteoporosis occupies first of metabolic bone disease.Bone
It is self-evident that the loose illness of matter, which brings patient and the pain of family,.According to statistics, the sclerotin that China about has more than 100,000,000 at present is dredged
Loose disease patient, it is contemplated that will be added to 200,000,000 1 million peoples to the year two thousand fifty.Cause the factor of more and more people's calcium deficiency：Life and
Work rhythm is accelerated, and operating pressure is big and life is irregular, the structure that is not careful in one's diet it is reasonably combined, daily calcium intake is less than
600 milligrams of persons are regarded as calcium insufficiency of intake, easily cause the shortage of internal calcium；Some a middle-aged person's work strains lack self and protect
Strong consciousness, thinks little of outdoor activity, so that the vitamin D synthesized in vivo is reduced, influences the absorption and utilization of calcium；Nutrition intake is not
Nutrient necessary to balanced or shortage human body.
The calcium agent of existing market is mostly the calcium of commercial synthesis.The calcium agent of commercial synthesis is with single nutrient component, is difficult to meet bone
Various nutritional needs, effect of supplemented calcium are undesirable during bone growth metabolism.Animal Bone is rich in a variety of mines such as calcium, phosphorus, iron, magnesium
Matter-element element, and it is similar to people's bone, and skeleton cell has stronger affinity to identical histocyte, thus utilization rate is more
It is high.
Chinese patent 201210382496.4 disclose it is a kind of using enzymatic hydrolysis and fermentation method extracted from animal skeleton
The technique of calcium element, bone calcium product made from this method is in good taste, the smell of no smelling of fish or mutton, and free calcium levels are higher.But this field
Technical staff knows, a kind of whether superior judgment criteria of calcium source, first is that calcium content is high, second is that calcium absorptivity is high.
In bone calcium made from above-mentioned technique, although free calcium levels increased after everfermentation, distance one excellent
Calcium source free calcium levels more or not high enough, the effect replenished the calcium is not satisfactory.Therefore it finds one kind and can either dramatically increase and mention
It takes free calcium levels in liquid and enables product that there is better effect of supplemented calcium, become technical staff's urgent problem.
Summary of the invention
The purpose of the present invention is to provide a kind of extraction processes of new osteocalcin, and the technique can be compared with the prior art
Preferably increase free calcium levels.
It is a further object of the present invention to provide a kind of calcium supplementing product with the market competitiveness, the product can be by people
Body preferably absorbs.
In order to achieve the above objectives, the present invention is realized using following technical proposal.
The present invention pre-processes the aggregate before enzymatic hydrolysis using the method for fermentation, then carries out enzymolysis and extraction calcium element again,
Then fermentation process is carried out again, and especially pretreated fermentation step is improved, selection including the kind to lactic acid bacteria,
Adding proportion of several lactic acid bacterias etc. is to reach finished product free calcium levels height and effect of supplemented calcium more preferably.
It is currently preferred that bone meal is broken to 15~25mm particle to enhance ferment effect, particle in 15mm or less and
25mm or more ferment effect is poor.Inventor passes through test of many times and experience, finally determines that 15~25mm granulometric range reaches
Calcium element ferment effect it is best.
Enzymatic hydrolysis primary fermentation of the present invention presses certain weight proportion using lactic acid bacteria and aggregate, 40~50 DEG C, ferments
12h.Concentration, fermentation temperature and the time of the weight proportion of lactic acid bacteria and aggregate, lactic acid bacteria in fermentation liquid are to final obtained
Free calcium levels have important influence in product, and the weight proportion and lactic acid bacteria of lactic acid bacteria and aggregate are dense in fermentation liquid
The influence spent to free calcium is maximum.Inventor determines that lactic acid bacteria accounts for aggregate after repetition test according to the working experience of oneself many years
Weight ratio is 0.03~0.07%, under conditions of aggregate and water w/v 1: 3~5, can reach optimal ferment effect.
Preferably, aggregate and water w/v are 1: 4, and it is 0.05% that lactic acid bacteria, which accounts for aggregate weight ratio, and fermentation temperature is
45℃。
The ferment effect that inventor has carried out a large amount of lactic acid bacterium strain selection and various lactobacillus is added in varing proportions
Test, has been surprisingly found that using bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and youth bifid
The effect that three kinds of lactic acid bacterias of bacillus (B.adolescentis) are fermented when being combined with weight ratio 1: 2: 2 is more preferable, relatively using cream
Bifidobacterium (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and Lactobacillus helveticus (L.helviticus) are with weight
Ferment effect when amount is combined than 1: 3: 2 has further promotion, and free calcium levels, which are compared, in enzymolysis liquid improves
30% or more.
Secondary fermentation of the present invention uses leuconostoc cremoris and citric acid, and between the two matching compares final produce
The taste of object, production efficiency all have important influence.Inventor finally determines that leuconostoc cremoris accounts for aggregate weight ratio and is
0.02~0.06%, under conditions of leuconostoc cremoris and citric acid weight ratio 1: 20~30, the final product effect of fermentation compared with
It is good.Under the above conditions, it determines that fermentation temperature is 39 DEG C, when fermentation time is 12h, can achieve the optimization of production efficiency.
Preferably, it is 0.03%, leuconostoc cremoris and citric acid weight ratio 1 that leuconostoc cremoris, which accounts for aggregate weight ratio:
Under conditions of 24, the final product effect of fermentation is best.
The present invention achievees the purpose that sterilizing using the method for high-temperature sterilization, while also removing the lactic acid in enzymolysis liquid
Bacterium.
Enzymatic hydrolysis of the invention uses secondary enzymolysis method, after screening, finally found that with pepsin and Papain
Enzyme, which is digested, has better effect compared with other protease.Optimal enzymolysis scheme is pH value 1 in extraction process of the invention
~3, pepsin is added, 35~40 DEG C of hydrolysis temperature, stirs 1~2h；PH value 6~7, addition papain, 40~50 DEG C,
Stir 1~2h.
Preferably, hydrolysis temperature is 38 DEG C after pepsin is added, and hydrolysis temperature is 45 DEG C after papain is added.
Two kinds of protease used in hydrolysis bone meal respectively account for aggregate weight ratio and are preferred for 0.04~0.08%.
Preferably, it is 0.06% that pepsin, which accounts for aggregate weight ratio, and it is 0.08% that papain, which accounts for aggregate weight ratio,.
Auxiliary material such as starch, dextrin, lactose, crystallite are fine on composition of the invention and any or more than one pharmacies
Tie up element, hypromellose, polyethylene glycol, magnesium stearate, superfine silica gel powder, xylitol, lactitol, glucose, glycine, sweet
The various dosage forms that dew alcohol, glycine etc. are mixed, for example, can be made into tablet, sustained release tablets, dripping pill, granule, capsule, powder
Agent, fine granule.Preferred dosage form is tablet or granule.
Osteocalcin made from the technical solution that technique of the present invention is 201210382496.4 compared with number of patent application is dissociated
Calcium content improves 30% or more.Final product obtained is in good taste, and calcium content is high, and the effect that is absorbed by the body is more preferable.
Test example
Beneficial effects of the present invention, all implementation of the present invention are further illustrated below by way of some specific experimental datas
Example can make with this similar in experiment effect, following data is merely illustrative.
One, product free calcium levels detect
1, materials and methods
1.1 ox bone：Hebei good fortune is purchased from into five rich food limited liability companies
1.2 samples 1：The sample obtained according to the embodiment 3 that number of patent application is 201210382496.4 using ox bone；Sample
Product 2：The sample obtained according to the embodiment 6 that number of patent application is 201210382496.4 using ox bone；Sample 3：According to this hair
The sample that bright embodiment 2 is obtained using ox bone；Sample 4：The sample obtained according to the embodiment of the present invention 3 using ox bone；Sample 5：
The sample obtained according to the embodiment of the present invention 4 using ox bone；Sample 6：The sample obtained according to the embodiment of the present invention 5 using ox bone
Product；Sample 7：The sample obtained according to the embodiment of the present invention 6 using ox bone；Sample 8：Ox bone is used according to the embodiment of the present invention 7
The sample of acquisition；Sample 9：The sample obtained according to the embodiment of the present invention 8 using ox bone；Sample 10：According to the embodiment of the present invention 9
The sample obtained using ox bone；Sample 11：The sample obtained according to the embodiment of the present invention 10 using ox bone；Sample 12：According to this
The sample that inventive embodiments 11 are obtained using ox bone.
1.3 method：Using atomic absorption spectroscopy determination free calcium levels.
2, result
Through date comprision, the free calcium levels of product obtained by the present invention have compared with sample 1, sample 2
Significant to improve, increase rate reaches 30.39%~44.06%.
Two, efficacy test
1 data and method
1.1 general information：
Total cases are 120, male 58, women 62, the age 42~54 years old, take aspiration for the product of our unit
Person, all cases are below normal value using the Dual-energy X-rays absorptionmetry measurement bone density of Lunar company of the U.S., are bone loss
Or osteoporosis.It is randomly divided into test A group, test B group, test C group and control A group, control B group, blank control group：Test A
Group 20, male 10, women 10, the age, average age was 47.2 years old at 43~51 years old；Test B group 20, male 9,
Women 11, the age, average age was 46.6 years old at 42~51 years old；Test C group 20, male 11, women 9, the age exists
44~50 years old, average age was 47.0 years old；Control A group 20, male 9, women 11, the age at 42~53 years old, average year
Age is 46.9 years old；Control B group 20, male 10, women 10, the age, average age was 45.8 years old at 42~50 years old；Blank
Control group 20, male 9, women 11, the age, average age was 46.4 years old at 44~51 years old.Six groups of volunteers the age,
Significant difference (P > 0.05) is not present in the general information such as the state of an illness, is comparable.
1.2 administrated method
Test A group gives the product obtained according to the embodiment of the present invention 2 using ox bone；Test B group is given according to the present invention
The product that embodiment 3 is obtained using ox bone；Test C group gives the product obtained according to the embodiment of the present invention 4 using ox bone；Control
A group gives a patent, and application No. is the products that 201210382496.4 embodiment 3 is obtained using ox bone；Control B group gives a patent
Application No. is the following products of product that 201210382496.4 embodiment 6 is obtained using ox bone；Blank control group, which is not taken, appoints
What calcium supplementing product only exhorts the high food of its more edible calcium content, gets sun more.In addition to blank control group, remaining 5 groups are taken production
The each 20g of product, 2 times a day.All cases are taken 3 months and are treated, and check bone density after the course for the treatment of.
1.3 observation items and measurement method
Bone density (BMD)：Respectively at using dual energy X-ray absorptiometry measuring instrument, (French Midlink company is raw before and after treatment
Produce) lumbar vertebrae normotopia (L1~L4) and femoral neck BMD value are measured, unit uses g/cm2。
1.4 statistical procedures
Measurement data mean ± standard deviationIt indicates, is examined using t for statistical analysis.
2 results
The pretherapy and post-treatment bone density value comparison of 2.1 6 groups of patients
1 six groups of table pretherapy and post-treatment BMD values compare (g/cm2,)
Note：* compared with before treatment, P < 0.05, * * are compared with before treatment, P < 0.01；
△ compared with the control group, P < 0.05.
3 conclusions
The embodiment of the present invention tests A group, test B group, test C group in terms of increasing bone density with the effect of highly significant
Fruit, P < 0.01；Have the effect of brilliant increase bone density, P < 0.05 compared with control A group, control B group.Inventor is to this
It invents all embodiments write to be tested according to above-mentioned test method, and has obtained similar test result,
It proves that osteocalcin made from preparation method of the present invention has effects that significantly to increase bone density, is with number of patent application
201210382496.4 osteocalcin is compared, and after not only free calcium levels are high, but also human body is taken, has good increase bone close
The effect of degree, i.e. absorptivity are high, good absorption effect.
Specific embodiment
Embodiment 1
Prepare culture medium：MRS culture medium：Peptone 1%, beef extract 1%, yeast extract 0.5%, glucose 2%, phosphoric acid hydrogen
Dipotassium 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 DEG C, sterilize 30min.
Embodiment 2
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 3, lactic acid bacteria account for aggregate weight
Than being 0.03%, 5% sucrose of total weight is added, 40 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 1, pepsin is added, accounting for aggregate weight ratio is 0.04%, 35 DEG C of hydrolysis temperature, is stirred
1h adds alkali to adjust pH value to 6, papain is added, accounting for aggregate weight ratio is 0.04%, 40 DEG C, stirs 1h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.02%, leuconostoc cremoris and citric acid weight ratio 1: 20, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 3
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 5, lactic acid bacteria account for aggregate weight
Than being 0.07%, 5% sucrose of total weight is added, 50 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 3, pepsin is added, accounting for aggregate weight ratio is 0.08%, 40 DEG C of hydrolysis temperature, is stirred
2h adds alkali to adjust pH value to 7, papain is added, accounting for aggregate weight ratio is 0.08%, 50 DEG C, stirs 2h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.06%, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 4
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 4, lactic acid bacteria account for aggregate weight
Than being 0.05%, 5% sucrose of total weight is added, 45 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 2, pepsin is added, accounting for aggregate weight ratio is 0.06%, 38 DEG C of hydrolysis temperature, is stirred
1.5h adds alkali to adjust pH value to 6, papain is added, accounting for aggregate weight ratio is 0.08%, 45 DEG C, stirs 1~2h, heating
Enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.03%, leuconostoc cremoris and citric acid weight ratio 1: 24, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 5
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 3, lactic acid bacteria account for aggregate weight
Than being 0.07%, 5% sucrose of total weight is added, 50 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 1, pepsin is added, accounting for aggregate weight ratio is 0.08%, 40 DEG C of hydrolysis temperature, is stirred
1h adds alkali to adjust pH value to 7, papain is added, accounting for aggregate weight ratio is 0.04%, 40 DEG C, stirs 2h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.02%, leuconostoc cremoris and citric acid weight ratio 1: 22, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 6
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 5, lactic acid bacteria account for aggregate weight
Than being 0.06%, 5% sucrose of total weight is added, 43 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 1~3, pepsin is added, accounting for aggregate weight ratio is 0.05%, 36 DEG C of hydrolysis temperature, is stirred
1h is mixed, adds alkali to adjust pH value to 6, papain is added, accounting for aggregate weight ratio is 0.06%, 48 DEG C, stirs 1h, heating is gone out
Enzyme；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.04%, leuconostoc cremoris and citric acid weight ratio 1: 28, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 7
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 5, lactic acid bacteria account for aggregate weight
Than being 0.07%, 5% sucrose of total weight is added, 46 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 3, pepsin is added, accounting for aggregate weight ratio is 0.08%, 37 DEG C of hydrolysis temperature, is stirred
2h adds alkali to adjust pH value to 6, papain is added, accounting for aggregate weight ratio is 0.04%, 40 DEG C, stirs 1h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.06%, leuconostoc cremoris and citric acid weight ratio 1: 25, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 8
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 4, lactic acid bacteria account for aggregate weight
Than being 0.06%, 5% sucrose of total weight is added, 50 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 2, pepsin is added, accounting for aggregate weight ratio is 0.07%, 35 DEG C of hydrolysis temperature, is stirred
1h adds alkali to adjust pH value to 7, papain is added, accounting for aggregate weight ratio is 0.04%, 45 DEG C, stirs 2h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.02%, leuconostoc cremoris and citric acid weight ratio 1: 27, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 9
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 3, lactic acid bacteria account for aggregate weight
Than being 0.05%, 5% sucrose of total weight is added, 40 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 3, pepsin is added, accounting for aggregate weight ratio is 0.06%, 40 DEG C of hydrolysis temperature, is stirred
1h adds alkali to adjust pH value to 7, papain is added, accounting for aggregate weight ratio is 0.06%, 50 DEG C, stirs 1h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.06%, leuconostoc cremoris and citric acid weight ratio 1: 20, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 10
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 4, lactic acid bacteria account for aggregate weight
Than being 0.03%, 5% sucrose of total weight is added, 46 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 1, pepsin is added, accounting for aggregate weight ratio is 0.04%, 40 DEG C of hydrolysis temperature, is stirred
2h adds alkali to adjust pH value to 6, papain is added, accounting for aggregate weight ratio is 0.08%, 45 DEG C, stirs 1h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.04%, leuconostoc cremoris and citric acid weight ratio 1: 30, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Embodiment 11
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 5, lactic acid bacteria account for aggregate weight
Than being 0.07%, 5% sucrose of total weight is added, 50 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 1, pepsin is added, accounting for aggregate weight ratio is 0.06%, 37 DEG C of hydrolysis temperature, is stirred
2h adds alkali to adjust pH value to 6, papain is added, accounting for aggregate weight ratio is 0.04%, 48 DEG C, stirs 1h, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for bone
Expect that weight ratio is 0.03%, leuconostoc cremoris and citric acid weight ratio 1: 25, fermentation temperature are 39 DEG C, fermentation time 12h,
Add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
Claims (10)
1. a kind of preparation method for the osteocalcin for extracting preparation by animal skeleton, its step are as follows：
(1) bone crushes, sterilizes；
(2) bifidobacterium lactis (B.lactis), lactobacillus bulgaricus (L.bulgaricus) and bifidobacterium adolescentis are taken
(B.adolescentis), three kinds of lactic acid bacteria weight ratios 1: 2: 2；Aggregate and water w/v 1: 3~5, lactic acid bacteria accounts for aggregate
Weight ratio is 0.03~0.07%, 5% sucrose of total weight is added, 40~50 DEG C, ferment 12h, 120 DEG C of sterilizing 30min；
(3) add hydrochloric acid tune pH value 1~3, pepsin is added, accounting for aggregate weight ratio is 0.04~0.08%, hydrolysis temperature 35~
40 DEG C, 1~2h is stirred, adds alkali to adjust pH value to 6~7, papain is added, accounting for aggregate weight ratio is 0.04~0.08%,
40~50 DEG C, 1~2h is stirred, heats enzyme deactivation；
(4) 5% glucose of total weight is added, leuconostoc cremoris is added and citric acid, leuconostoc cremoris account for aggregate weight
Amount is than being 0.02~0.06%, and leuconostoc cremoris and citric acid weight ratio 1: 20~30, fermentation temperature are 39 DEG C, fermentation time
For 12h, add alkali neutralization, 120 DEG C of sterilizings are drying to obtain.
2. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that bone in preparation methods steps (1)
It is crushed to 15~25mm particle.
3. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that aggregate in preparation methods steps (2)
It is 1 ︰ 4 with water w/v, it is 0.05% that lactic acid bacteria, which accounts for aggregate weight ratio,.
4. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that fermentation in preparation methods steps (2)
Temperature is 45 DEG C.
5. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that be added in preparation methods steps (3)
Pepsin account for aggregate weight ratio be 0.06%, the papain of addition account for aggregate weight ratio be 0.08%.
6. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that be added in preparation methods steps (3)
Hydrolysis temperature is 38 DEG C after pepsin, and hydrolysis temperature is 45 DEG C after papain is added.
7. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that be added in preparation methods steps (4)
Leuconostoc cremoris account for aggregate weight ratio be 0.03%.
8. a kind of preparation method of osteocalcin described in claim 1, which is characterized in that butterfat in preparation methods steps (4)
Leukonid and citric acid weight ratio 1: 24.
9. a kind of osteocalcin as made from claim 1~8 any claim preparation method.
10. a kind of application of osteocalcin as claimed in claim 9 in the drug and health food that preparation increases bone density function.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102860523A (en) * | 2012-10-10 | 2013-01-09 | 天狮集团有限公司 | Bone calcium extraction method by double fermentation and bone gla protein |
| CN102871123A (en) * | 2012-10-10 | 2013-01-16 | 天狮集团有限公司 | Method for preparing bone calcium and bone gla protein |
| CN102871126A (en) * | 2012-10-10 | 2013-01-16 | 天狮集团有限公司 | Bone calcium extraction method and osteocalcin |
| CN103211836A (en) * | 2013-04-11 | 2013-07-24 | 天津天狮生物发展有限公司 | Composition for treating osteoporosis |
| 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 |
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2018
- 2018-05-04 CN CN201810418980.5A patent/CN108850421A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102860523A (en) * | 2012-10-10 | 2013-01-09 | 天狮集团有限公司 | Bone calcium extraction method by double fermentation and bone gla protein |
| CN102871123A (en) * | 2012-10-10 | 2013-01-16 | 天狮集团有限公司 | Method for preparing bone calcium and bone gla protein |
| CN102871126A (en) * | 2012-10-10 | 2013-01-16 | 天狮集团有限公司 | Bone calcium extraction method and osteocalcin |
| CN103211836A (en) * | 2013-04-11 | 2013-07-24 | 天津天狮生物发展有限公司 | Composition for treating osteoporosis |
| 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 |
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Application publication date: 20181123 |