CN105768111A - Preparing method and application of ocean polypeptide-calcium chelate - Google Patents
Preparing method and application of ocean polypeptide-calcium chelate Download PDFInfo
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- CN105768111A CN105768111A CN201610234612.6A CN201610234612A CN105768111A CN 105768111 A CN105768111 A CN 105768111A CN 201610234612 A CN201610234612 A CN 201610234612A CN 105768111 A CN105768111 A CN 105768111A
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- ocean
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- calcium
- calcium chelate
- chelate
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- 238000000034 method Methods 0.000 title claims abstract description 11
- 241001465754 Metazoa Species 0.000 claims abstract description 29
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 28
- 229920001184 polypeptide Polymers 0.000 claims abstract description 26
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 26
- 238000002360 preparation method Methods 0.000 claims abstract description 22
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 15
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 15
- 108091005804 Peptidases Proteins 0.000 claims abstract description 12
- 239000004365 Protease Substances 0.000 claims abstract description 12
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 12
- 239000000047 product Substances 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims abstract description 10
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 9
- 102000004190 Enzymes Human genes 0.000 claims abstract description 8
- 108090000790 Enzymes Proteins 0.000 claims abstract description 8
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- 239000007788 liquid Substances 0.000 claims abstract description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 241000195474 Sargassum Species 0.000 claims description 16
- 230000033228 biological regulation Effects 0.000 claims description 16
- 241000251468 Actinopterygii Species 0.000 claims description 13
- 238000001035 drying Methods 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 10
- 241000371997 Eriocheir sinensis Species 0.000 claims description 9
- 241000237502 Ostreidae Species 0.000 claims description 9
- 235000015097 nutrients Nutrition 0.000 claims description 9
- 235000020636 oyster Nutrition 0.000 claims description 9
- 239000013522 chelant Substances 0.000 claims description 7
- 239000002775 capsule Substances 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 6
- 235000013305 food Nutrition 0.000 claims description 6
- 239000008187 granular material Substances 0.000 claims description 6
- 239000012535 impurity Substances 0.000 claims description 6
- 230000004044 response Effects 0.000 claims description 6
- 238000005204 segregation Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 230000002459 sustained effect Effects 0.000 claims description 6
- 239000003826 tablet Substances 0.000 claims description 6
- 108090000145 Bacillolysin Proteins 0.000 claims description 5
- 108091005658 Basic proteases Proteins 0.000 claims description 5
- 102000035092 Neutral proteases Human genes 0.000 claims description 5
- 108091005507 Neutral proteases Proteins 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 5
- 238000003916 acid precipitation Methods 0.000 claims description 5
- 239000003513 alkali Substances 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 239000008188 pellet Substances 0.000 claims description 5
- 238000001556 precipitation Methods 0.000 claims description 5
- 230000006920 protein precipitation Effects 0.000 claims description 5
- 238000010298 pulverizing process Methods 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 239000000284 extract Substances 0.000 claims description 4
- 239000002552 dosage form Substances 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 abstract description 16
- 229910052791 calcium Inorganic materials 0.000 abstract description 16
- 239000011575 calcium Substances 0.000 abstract description 16
- 150000001413 amino acids Chemical class 0.000 abstract description 5
- 231100000252 nontoxic Toxicity 0.000 abstract description 4
- 230000003000 nontoxic effect Effects 0.000 abstract description 4
- 239000001110 calcium chloride Substances 0.000 abstract description 3
- 229910001628 calcium chloride Inorganic materials 0.000 abstract description 3
- 235000021190 leftovers Nutrition 0.000 abstract 3
- 235000015872 dietary supplement Nutrition 0.000 abstract 2
- 229940069978 calcium supplement Drugs 0.000 abstract 1
- 230000009920 chelation Effects 0.000 abstract 1
- 230000009849 deactivation Effects 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- 238000002955 isolation Methods 0.000 abstract 1
- 239000002002 slurry Substances 0.000 abstract 1
- 230000001502 supplementing effect Effects 0.000 abstract 1
- 230000007723 transport mechanism Effects 0.000 abstract 1
- 229960005069 calcium Drugs 0.000 description 14
- 235000018102 proteins Nutrition 0.000 description 11
- 241000238557 Decapoda Species 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 238000013459 approach Methods 0.000 description 3
- 239000007910 chewable tablet Substances 0.000 description 3
- 229940068682 chewable tablet Drugs 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 229940106635 calcium amino acid chelate Drugs 0.000 description 2
- 159000000007 calcium salts Chemical class 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- UOXSXMSTSYWNMH-UHFFFAOYSA-L zinc;2-aminoacetate Chemical compound [Zn+2].NCC([O-])=O.NCC([O-])=O UOXSXMSTSYWNMH-UHFFFAOYSA-L 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 206010006956 Calcium deficiency Diseases 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 description 1
- 239000001354 calcium citrate Substances 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 239000001527 calcium lactate Substances 0.000 description 1
- 229960002401 calcium lactate Drugs 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 235000013337 tricalcium citrate Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
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
- A23J1/001—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste
- A23J1/002—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste from animal waste materials
-
- 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
- A23J1/001—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste
- A23J1/005—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste from vegetable waste materials
-
- 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
- A23J1/006—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from vegetable materials
-
- 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
- A23J1/04—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from fish or other sea animals
-
- 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
- A23J3/30—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
- 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
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nutrition Science (AREA)
- Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a preparing method and application of an ocean polypeptide-calcium chelate.The preparing method includes the steps that protein in ocean animals and plants and leftovers thereof is extracted through an alkali-solution and acid-isolation method, or the ocean animals and plants and leftovers thereof are smashed and ground into thick liquid, protease is added into obtained protein or slurry to conduct enzymolysis, enzyme deactivation is carried out, and an ocean polypeptide compound solution is prepared; calcium in inorganic calcium chloride and ocean polypeptide are chelated to obtain the ocean polypeptide-calcium chelate, and the obtained ocean polypeptide-calcium chelate can be further prepared into leisure food, or dietary supplements or drugs.The preparation process is simple, the obtained ocean polypeptide-calcium chelate and products thereof have a unique chelation system and transport mechanism, are safe, nontoxic, easy to absorb and capable of supplementing amino acid and calcium at the same time, is the first choice of a calcium supplement and provides a new ideal for application of the ocean animals and plants and the leftovers thereof.
Description
Technical field
The invention belongs to biological technical field, be specifically related to preparation method and the application of a kind of ocean polypeptide-calcium chelate.
Background technology
Calcium is one of human essential elements, participates in the multiple physiological activity of body.Calcium deficiency can cause human body numerous disease, makes human body enter sub-health state, therefore, replenishes the calcium and become the topic that people pay close attention to for a long time.Traditional inorganic calcium salt, such as calcium carbonate, calcium phosphate, calcium chloride etc., has certain destruction to vitamin, and biological value is relatively low；Organic calcium salt, such as calcium citrate, calcium lactate, calisanin etc., although calcium absorptivity increases, but its dissolubility is big, readily soluble mistake, and price is high.In recent years, calcium amino acid chelate owing to easily being absorbed, safety non-toxic, price are low, can supplement aminoacid and calcium simultaneously and become first-selection of replenishing the calcium.But, research shows, polypeptide chelate calcium is due to the chelating system of its uniqueness and transporting mechanism so that it is is more readily absorbed than calcium amino acid chelate and has faster infiltration rate.Therefore, the polypeptide chelate calcium having certain biological function has wide development space.
Containing substantial amounts of protein in the marine animal and plants such as Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, sea oyster and leftover bits and pieces thereof, it is to prepare various active polypeptide and amino acid whose good raw material.The present invention utilizes biotechnology that the marine animal and plants such as Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, sea oyster and leftover bits and pieces thereof are carried out the deep processing of protein resource, carry out the screening of biologically active peptide and extract separation, then calcium chloride is utilized to chelate, preparation ocean polypeptide-calcium chelate.This marine bioactivity polypeptide-calcium chelate is good at different acid-base condition stability inferiors, absorbance and the utilization rate of calcium constituent can be significantly improved, but also there is antioxidation, antibacterial, immunomodulating, blood fat reducing and blood sugar lowering isoreactivity, there are the highest Development volue and application prospect, Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, the sea marine animal and plant such as oyster, clam and the added value of leftover bits and pieces thereof can be improved simultaneously, utilization for Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, the sea marine animal and plant such as oyster, clam and leftover bits and pieces resource thereof opens new approach, has wide market prospect.
Summary of the invention
It is an object of the invention to provide preparation method and the application of a kind of ocean polypeptide-calcium chelate, technological operation is easy, safety is high advantage that it has.
For achieving the above object, the present invention adopts the following technical scheme that
A kind of ocean polypeptide-calcium chelate, its preparation method comprises the steps:
(1) preparation of ocean polypeptide: utilize alkali extraction-acid precipitation to extract the albumen in marine animal and plant and leftover bits and pieces thereof, or directly by after marine animal and plant and leftover bits and pieces pulverizing, defibrination, in gained albumen or serosity, add protease carry out enzymolysis, enzyme denaturing, make ocean polypeptide complex solution；
(2) preparation of ocean polypeptide-calcium chelate: chelate liquid is legal that gained ocean polypeptide complex solution is prepared as ocean polypeptide-calcium chelate in application, and gained ocean polypeptide-calcium chelate is carried out lyophilization or cold drying.
Described marine animal and plant includes Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, sea oyster, clam.
During in step (1), alkali extraction-acid precipitation extracts marine animal and plant and leftover bits and pieces thereof, the concrete operations of albumen are: first by marine animal and plant and leftover bits and pieces micronizing thereof, then marine animal and plant and leftover bits and pieces thereof after pulverizing are mixed by mass volume ratio 1:10-50 with water, utilize the aqueous slkali regulation pH to 8.0-11.0 of 0.5-2wt%, after supersound extraction 10-60min, 60-120min is extracted in 50-100 DEG C of water-bath, filter, filtrate carries out albumen precipitation with salt acid for adjusting pH to 2.5-4.0, 3000-6000r/min is centrifuged 5-10min and obtains protein precipitation, it is washed to recentrifuge after neutrality, gained pellet frozen is drying to obtain the albumen in marine animal and plant and leftover bits and pieces thereof；
Described aqueous slkali is NaOH solution or KOH solution.
Enzymolysis described in step (1) is water use regulation gained albumen or the protein concentration of serosity, make protein concentration reach 1.0-5.0wt%, and regulate pH value of solution to 7.0-11.0, then keep the temperature at 40-60 DEG C, add the protease of 1.0-5.0wt%, hydrolyze 30-240min.Described protease is alkaline protease, neutral protease or compound protease.
The concrete operations of the polypeptide of chelate liquid legal preparation ocean described in step (2)-calcium chelate are: step (1) gained ocean polypeptide complex solution 1-6:6 by volume is added 0.5-2mol/L calcium chloride solution, stir, regulation pH value of solution is 3.0-9.0, cool down after sustained response 1.0-3.0h under the conditions of 30-80 DEG C, the removal of impurity is gone in centrifugation, it is subsequently adding the ethanol of supernatant volume 3-5 times, 75-95wt%, stand 12-24h, centrifugal segregation supernatant, it is thus achieved that ocean polypeptide-calcium chelate.
Described in step (2), the temperature of cold drying is≤65 DEG C.
Gained ocean polypeptide-calcium chelate technique can be fabricated to leisure food, nutrient and healthcare products or medicine routinely further；
Described nutrient and healthcare products or medicine are pharmaceutically acceptable dosage form, including granule, capsule, tablet.
The present invention compared with prior art, has the advantage that
(1) albumen during alkali extraction-acid precipitation can extract marine animal and plant and leftover bits and pieces thereof, its extraction ratio is high, and the most less document about marine animal and plant protein extraction is reported.
(2) present invention is by controlling proteolysis time, can effectively control the palliating degradation degree of albumen, obtain the biologically active peptide with high calcium sequestering activity.
(3) present invention is by chelating gained ocean polypeptide with the calcium in inorganic chlorinating calcium, organic marine biologically active peptide-calcium chelate prepared and products thereof has chelating system and the transporting mechanism of uniqueness, safety non-toxic, easily absorbed, aminoacid and calcium ion can be supplemented simultaneously, and cheap.
(4) present invention is that Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, the sea marine animal and plant such as oyster, clam and the application of leftover bits and pieces thereof provide new approaches, provides theoretical foundation and technical support for exploitation organic calcium supplementary with functional food.
Detailed description of the invention
The preparation method of a kind of ocean polypeptide-calcium chelate, comprises the steps:
(1) preparation of ocean polypeptide: first by Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, sea oyster, the marine animal and plants such as clam and leftover bits and pieces thereof carry out micronizing, then will pulverize after marine animal and plant and leftover bits and pieces with water by mass volume ratio 1:10-50(g/mL) mix, utilize NaOH solution or the KOH solution regulation pH to 8.0-11.0 of 0.5-2wt%, after supersound extraction 10-60min, 60-120min is extracted in 50-100 DEG C of water-bath, filter, filtrate carries out albumen precipitation with salt acid for adjusting pH to 2.5-4.0, 3000-6000r/min is centrifuged 5-10min and obtains protein precipitation, it is washed to recentrifuge after neutrality, gained pellet frozen is drying to obtain the albumen in marine animal and plant and leftover bits and pieces thereof；Then gained albumen is dissolved in water, protein concentration is made to reach 1.0-5.0wt%, and regulate pH value of solution to 7.0-11.0, then 40-60 DEG C is kept the temperature at, add the alkaline protease of 1.0-5.0wt%, neutral protease or compound protease, hydrolysis 30-240min, then through enzyme denaturing, make ocean polypeptide complex solution；
Or directly by after Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, the sea marine animal and plant such as oyster, clam and leftover bits and pieces pulverizing, defibrination, its protein concentration is made to reach 1.0-5.0wt%, and regulate pH value of solution to 7.0-11.0, then 40-60 DEG C is kept the temperature at, add the alkaline protease of 1.0-5.0wt%, neutral protease or compound protease, hydrolysis 30-240min, then through enzyme denaturing, make ocean polypeptide complex solution；
(2) preparation of ocean polypeptide-calcium chelate: step (1) gained ocean polypeptide complex solution 1-6:6 by volume is added 0.5-2mol/L calcium chloride solution, stir, regulation pH value of solution is 3.0-9.0, cool down after sustained response 1.0-3.0h under the conditions of 30-80 DEG C, the removal of impurity is gone in centrifugation, it is subsequently adding the ethanol of supernatant volume 3-5 times, 75-95wt%, stand 12-24h, centrifugal segregation supernatant, gained precipitates freeze-dried or less than 65 DEG C cold drying, it is thus achieved that ocean polypeptide-calcium chelate.
Gained ocean polypeptide-calcium chelate technique can be fabricated to leisure food, nutrient and healthcare products or medicine routinely further；Described nutrient and healthcare products or medicine are pharmaceutically acceptable dosage form, including granule, capsule, tablet.
In order to make content of the present invention easily facilitate understanding, below in conjunction with detailed description of the invention, technical solutions according to the invention are described further, but the present invention is not limited only to this.
Embodiment 1
(1) first by after Thallus Laminariae (Thallus Eckloniae) micronizing, with water by mass volume ratio 1:30(g/mL) mix, utilizing the NaOH solution regulation pH to 9.0 of 0.5wt%, after supersound extraction 30min, 100min is extracted in 60 DEG C of water-baths, by filtered through gauze after extraction, filtrate carries out albumen precipitation with salt acid for adjusting pH to 3.0, and 4000r/min is centrifuged 10min and obtains protein precipitation, is washed to recentrifuge after neutrality, gained pellet frozen is drying to obtain Thallus Laminariae (Thallus Eckloniae) albumen, its extraction rate reached 74.16%；
(2) gained Thallus Laminariae (Thallus Eckloniae) albumen is dissolved in water, makes protein concentration reach 3.0wt%, with NaOH solution regulation pH value of solution to 9.0, then keep the temperature at 50 DEG C, add the neutral protease of 3.0wt%, hydrolyze 90min, then through enzyme denaturing, make Thallus Laminariae (Thallus Eckloniae) polypeptide complex solution；
(3) gained Thallus Laminariae (Thallus Eckloniae) polypeptide complex solution 2:6 by volume is added 1mol/L calcium chloride solution, stir, regulation pH value of solution is 8.0, cooling down after sustained response 2.0h under the conditions of 40 DEG C, centrifugation is gone the removal of impurity, is subsequently adding supernatant volume 4 times, the ethanol of 85wt%, stand 20h, centrifugal segregation supernatant, gained precipitates through cold drying, it is thus achieved that Thallus Laminariae (Thallus Eckloniae) polypeptide-calcium chelate；
(4) prepared Thallus Laminariae (Thallus Eckloniae) polypeptide-calcium chelate is fabricated to various leisure food further by existing common process, or the nutrient and healthcare products of the form such as granule, capsule, tablet (chewable tablet) or medicine.
Embodiment 2
A kind of method preparing ocean polypeptide-calcium chelate, comprises the steps:
(1) after small fish and shrimp being pulverized, with water by mass volume ratio 1:10(g/mL) mix defibrination, protein concentration is made to reach 1wt%, and with NaOH solution regulation pH value of solution to 7.0, then keep the temperature at 60 DEG C, add the compound protease of 1.0wt%, hydrolyze 240min, again through enzyme denaturing, make small fish and shrimp polypeptide complex solution；
(2) step (1) gained ocean polypeptide complex solution 1:6 by volume is added 0.5mol/L calcium chloride solution, stir, regulation pH value of solution is 3.0, cooling down after sustained response 3.0h under the conditions of 30 DEG C, centrifugation is gone the removal of impurity, is subsequently adding supernatant volume 3 times, the ethanol of 75wt%, stand 12h, centrifugal segregation supernatant, gained precipitates through cold drying, it is thus achieved that small fish and shrimp polypeptide-calcium chelate；
(3) prepared fish and shrimp polypeptide-calcium chelate is fabricated to various leisure food further by existing common process, or the nutrient and healthcare products of the form such as granule, capsule, tablet (chewable tablet) or medicine.
Embodiment 3
(1) first by after Sargassum micronizing, with water by mass volume ratio 1:50(g/mL) mix, utilizing the KOH solution regulation pH to 11.0 of 2wt%, after supersound extraction 60min, 60min is extracted in 100 DEG C of water-baths, by filtered through gauze after extraction, filtrate carries out albumen precipitation with salt acid for adjusting pH to 4.0, and 6000r/min is centrifuged 5min and obtains protein precipitation, is washed to recentrifuge after neutrality, gained pellet frozen is drying to obtain Sargassum protein, its extraction rate reached 76.43%；
(2) gained Sargassum protein is dissolved in water, makes protein concentration reach 5.0wt%, with NaOH solution regulation pH value of solution to 11.0, then keep the temperature at 60 DEG C, add the alkaline protease of 5.0wt%, hydrolyze 240min, again through enzyme denaturing, make Sargassum polypeptide complex solution；
(3) gained Sargassum polypeptide complex solution 1:1 by volume is added 2mol/L calcium chloride solution, stir, regulation pH value of solution is 9.0, cooling down after sustained response 1.0h under the conditions of 80 DEG C, centrifugation is gone the removal of impurity, is subsequently adding supernatant volume 5 times, the ethanol of 95wt%, stand 16h, centrifugal segregation supernatant, gained precipitates through cold drying, it is thus achieved that Sargassum polypeptide-calcium chelate；
(4) prepared Sargassum polypeptide-calcium chelate is fabricated to various leisure food further by existing common process, or the nutrient and healthcare products of the form such as granule, capsule, tablet (chewable tablet) or medicine.
Gained ocean of the present invention polypeptide-calcium chelate and products thereof has chelating system and the transporting mechanism of uniqueness, safety non-toxic, easily absorbed, aminoacid and calcium can be supplemented simultaneously, can the first-selection of calcium complement agent, the application for marine animal and plant and leftover bits and pieces thereof provides new approaches.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent and modification, all should belong to the covering scope of the present invention.
Claims (9)
1. the preparation method of ocean polypeptide-calcium chelate, it is characterised in that: comprise the steps:
(1) preparation of ocean polypeptide: utilize alkali extraction-acid precipitation to extract the albumen in marine animal and plant and leftover bits and pieces thereof, or directly by after marine animal and plant and leftover bits and pieces pulverizing, defibrination, in gained albumen or serosity, add protease carry out enzymolysis, enzyme denaturing, make ocean polypeptide complex solution；
(2) preparation of ocean polypeptide-calcium chelate: chelate liquid is legal that gained ocean polypeptide complex solution is prepared as ocean polypeptide-calcium chelate in application, and gained ocean polypeptide-calcium chelate is carried out lyophilization or cold drying.
The preparation method of ocean polypeptide-calcium chelate the most according to claim 1, it is characterised in that: described marine animal and plant includes Sargassum, fish, shrimp Eriocheir sinensis, Carnis Haliotidis, sea oyster, clam.
nullThe preparation method of ocean polypeptide-calcium chelate the most according to claim 1，It is characterized in that: during in step (1), alkali extraction-acid precipitation extracts marine animal and plant and leftover bits and pieces thereof, the concrete operations of albumen are: first by marine animal and plant and leftover bits and pieces micronizing thereof，Then marine animal and plant and leftover bits and pieces thereof after pulverizing are mixed by mass volume ratio 1:10-50 with water，Utilize the aqueous slkali regulation pH to 8.0-11.0 of 0.5-2wt%，After supersound extraction 10-60min，60-120min is extracted in 50-100 DEG C of water-bath，Filter，Filtrate carries out albumen precipitation with salt acid for adjusting pH to 2.5-4.0，3000-6000r/min is centrifuged 5-10min and obtains protein precipitation，It is washed to recentrifuge after neutrality，Gained pellet frozen is drying to obtain the albumen in marine animal and plant and leftover bits and pieces thereof；
Described aqueous slkali is NaOH solution or KOH solution.
The preparation method of ocean polypeptide-calcium chelate the most according to claim 1, it is characterized in that: protein concentration during enzymolysis is water use regulation gained albumen or serosity described in step (1), protein concentration is made to reach 1.0-5.0wt%, and regulate pH value of solution to 7.0-11.0, then 40-60 DEG C is kept the temperature at, add the protease of 1.0-5.0wt%, hydrolyze 30-240min.
5. according to the preparation method of ocean polypeptide-calcium chelate described in claim 1 or 4, it is characterised in that: described protease is alkaline protease, neutral protease or compound protease.
The preparation method of ocean polypeptide-calcium chelate the most according to claim 1, it is characterized in that: the concrete operations of the polypeptide of chelate liquid legal preparation ocean described in step (2)-calcium chelate are: step (1) gained ocean polypeptide complex solution 1-6:6 by volume is added 0.5-2mol/L calcium chloride solution, stir, regulation pH value of solution is 3.0-9.0, cool down after sustained response 1.0-3.0h under the conditions of 30-80 DEG C, the removal of impurity is gone in centrifugation, it is subsequently adding supernatant volume 3-5 times, the ethanol of 75-95wt%, stand 12-24h, centrifugal segregation supernatant, obtain ocean polypeptide-calcium chelate.
The preparation method of ocean polypeptide-calcium chelate the most according to claim 1, it is characterised in that: described in step (2), the temperature of cold drying is≤65 DEG C.
8. ocean polypeptide-calcium chelate that a method as claimed in claim 1 prepares.
9. the application of ocean as claimed in claim 8 polypeptide-calcium chelate, it is characterised in that: described ocean polypeptide-calcium chelate technique routinely is fabricated to leisure food, nutrient and healthcare products or medicine further；
Described nutrient and healthcare products or medicine are pharmaceutically acceptable dosage form, including granule, capsule, tablet.
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