CN114601014B - Preparation method of casein micelle concentrated solution with high thermal stability and low viscosity - Google Patents
Preparation method of casein micelle concentrated solution with high thermal stability and low viscosity Download PDFInfo
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- CN114601014B CN114601014B CN202210252172.2A CN202210252172A CN114601014B CN 114601014 B CN114601014 B CN 114601014B CN 202210252172 A CN202210252172 A CN 202210252172A CN 114601014 B CN114601014 B CN 114601014B
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- 239000005018 casein Substances 0.000 title claims abstract description 225
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 title claims abstract description 225
- 235000021240 caseins Nutrition 0.000 title claims abstract description 225
- 239000000693 micelle Substances 0.000 title claims abstract description 152
- 238000002360 preparation method Methods 0.000 title claims abstract description 36
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- 108010076119 Caseins Proteins 0.000 claims abstract description 245
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- 238000004132 cross linking Methods 0.000 claims abstract description 77
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 69
- -1 citrate ions Chemical class 0.000 claims abstract description 38
- 239000001509 sodium citrate Substances 0.000 claims abstract description 23
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims abstract description 23
- 229940038773 trisodium citrate Drugs 0.000 claims abstract description 23
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- 238000012545 processing Methods 0.000 claims abstract description 5
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- 230000000694 effects Effects 0.000 claims description 23
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- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 abstract description 29
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- 230000002195 synergetic effect Effects 0.000 abstract description 9
- 239000000243 solution Substances 0.000 description 98
- 239000000843 powder Substances 0.000 description 18
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 14
- 229910001424 calcium ion Inorganic materials 0.000 description 14
- 238000001914 filtration Methods 0.000 description 13
- 239000008367 deionised water Substances 0.000 description 12
- 229910021641 deionized water Inorganic materials 0.000 description 12
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 11
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 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 5
- 238000010494 dissociation reaction Methods 0.000 description 5
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- 238000006703 hydration reaction Methods 0.000 description 5
- 239000008101 lactose Substances 0.000 description 5
- 238000001471 micro-filtration Methods 0.000 description 5
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 4
- 102000007544 Whey Proteins Human genes 0.000 description 4
- 108010046377 Whey Proteins Proteins 0.000 description 4
- 239000011575 calcium Substances 0.000 description 4
- 229910052791 calcium Inorganic materials 0.000 description 4
- 229920001429 chelating resin Polymers 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 235000013336 milk Nutrition 0.000 description 4
- 239000008267 milk Substances 0.000 description 4
- 210000004080 milk Anatomy 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 239000003708 ampul Substances 0.000 description 3
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- 238000001694 spray drying Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 239000005862 Whey Substances 0.000 description 2
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- 239000013522 chelant Substances 0.000 description 2
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- 239000007788 liquid Substances 0.000 description 2
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- 238000005374 membrane filtration Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 229940080237 sodium caseinate Drugs 0.000 description 2
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- 238000000108 ultra-filtration Methods 0.000 description 2
- 235000021119 whey protein Nutrition 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000282828 Camelus bactrianus Species 0.000 description 1
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- 102000008934 Muscle Proteins Human genes 0.000 description 1
- 108010074084 Muscle Proteins Proteins 0.000 description 1
- 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 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 108060008539 Transglutaminase Proteins 0.000 description 1
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- OWMVSZAMULFTJU-UHFFFAOYSA-N bis-tris Chemical compound OCCN(CCO)C(CO)(CO)CO OWMVSZAMULFTJU-UHFFFAOYSA-N 0.000 description 1
- 230000037180 bone health Effects 0.000 description 1
- 150000005693 branched-chain amino acids Chemical class 0.000 description 1
- 235000020246 buffalo milk 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
- 235000020248 camel milk Nutrition 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 235000020247 cow milk Nutrition 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 235000020250 donkey milk Nutrition 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 238000005189 flocculation Methods 0.000 description 1
- 230000016615 flocculation Effects 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 108010010779 glutamine-pyruvate aminotransferase Proteins 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 235000020252 horse milk Nutrition 0.000 description 1
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 235000004213 low-fat Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000005974 protein supplement Nutrition 0.000 description 1
- 229940116540 protein supplement Drugs 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000012465 retentate Substances 0.000 description 1
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- 230000001953 sensory effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000020254 sheep milk Nutrition 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 102000003601 transglutaminase Human genes 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
- 239000003643 water by type Substances 0.000 description 1
- 235000020255 yak milk Nutrition 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
- A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
- A23J1/202—Casein or caseinates
-
- 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
- A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
- A23J1/205—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey from whey, e.g. lactalbumine
-
- 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
- A23J3/08—Dairy 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
- A23J3/04—Animal proteins
- A23J3/08—Dairy proteins
- A23J3/10—Casein
-
- 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
- A23J3/343—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy 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
- A23J3/343—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins
- A23J3/344—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins of casein
Abstract
The invention discloses a preparation method of casein micelle concentrated solution with high thermal stability and low viscosity, and belongs to the technical field of dairy product processing. The invention adopts MCC or MPC to prepare micelle complex solution, carries out Tg enzyme crosslinking treatment on the complex solution, and then adds citrate ions, or carries out decalcification treatment, or adds sodium ions; the Tg enzyme treatment achieves a kappa-casein crosslinking degree of 30% -90%; the concentration of the citrate ion added is 8-22mM/100g casein; the decalcification rate achieved by the decalcification treatment is 9% -25%; the citrate ions are carried in by trisodium citrate and citric acid, and the molar ratio of the trisodium citrate to the citric acid is 7.5:2.5-8.5:1.5. The invention has synergistic effect between Tg enzyme treatment and citrate ion addition, between Tg enzyme treatment and decalcification treatment and between Tg enzyme treatment and sodium ion addition, can better maintain the natural polymeric structure of micelle, and can inhibit aggregation among the micelle in the high-temperature sterilization and subsequent storage of the concentrated solution, so that the concentrated solution has higher thermal stability and lower viscosity.
Description
Technical Field
The invention relates to a preparation method of casein micelle concentrated solution with high thermal stability and low viscosity, belonging to the technical field of dairy product processing.
Background
Special groups such as the old, the weak, the patients, the sports and the health care require to take more high-quality protein so as to meet the nutrition requirement of organisms, promote the synthesis of muscle protein and further improve the quality of life. Among the numerous protein supplements, protein concentrate products have a high market potential, with protein contents of typically 4.2% and above, and small intake of these products can meet the body's high protein requirements, thus reducing the consumer's dietary burden.
The milk protein belongs to high-quality protein, has higher content of essential amino acid and branched chain amino acid, and is easy to digest and absorb. The common milk protein ingredients comprise whey protein and sodium caseinate, and protein molecules exist in a structure form of a monomer and an oligomer, so that the concentrated solution has thick taste and poor drinkability. The concentrated micelle-state casein (MCC) and the concentrated Milk Protein (MPC) are milk protein ingredients prepared by membrane filtration and separation, and are rich in casein micelles, and the natural structure of the casein micelles is kept better. The casein micelle is a spherical polymer formed by associating casein and colloidal calcium phosphate, and the particle size distribution range is 50-800nm. The natural structure of casein micelle, i.e. the copolymerization state of casein and calcium, makes the viscosity of the concentrated solution lower, and is easy for consumers to drink. The bioavailability of calcium ions in casein micelles is also high, which is helpful for improving bone health. In addition, MCC and MPC have the characteristics of high protein, low fat and low lactose content, and have better application prospects in the research and development and production of protein concentrate.
In the membrane filtration process of MCC and MPC, free components such as lactose, chelating agent, free ions and the like are gradually removed along with permeate, so that the colloidal stability of casein micelles is reduced. Protein concentrate produced by MCC and MPC is easy to aggregate during high-temperature sterilization treatment and subsequent storage, and causes instability phenomena such as precipitation, gel and the like, so that the heat transfer efficiency of sterilization equipment is reduced, and the sensory quality of the product is adversely affected.
Disclosure of Invention
[ technical problem ]
The protein concentrate produced by adopting traditional milk protein ingredients such as whey protein or sodium caseinate has thick taste and poor drinkability; protein concentrate produced by MCC or MPC rich in casein micelle is easy to produce instability phenomena such as precipitation, gel and the like in high-temperature sterilization treatment and subsequent storage.
The invention aims to solve the technical problems that: provided is a method for producing a casein micelle concentrate having high heat stability and low viscosity, which can effectively inhibit the generation of precipitates or gels during high-temperature sterilization and subsequent storage of the concentrate, and which can give a concentrate having a low viscosity and which is easy to drink.
Technical scheme
In order to solve the problems, the invention adopts MCC or MPC to prepare casein micelle complex solution; performing Tg enzyme (glutamine transaminase) treatment on the complex solution, crosslinking kappa-casein on the surface of the micelle, and inhibiting dissociation of kappa-casein from the surface of the micelle and calcium ion-induced aggregation among the micelles during the heat treatment; or citrate ions are added to moderately chelate free calcium ions, so that aggregation among micelles induced by the calcium ions is weakened; or moderately decalcifying the complex solution to weaken aggregation among the micelles induced by calcium ions; or sodium ions are added to moderately shield the negative charges carried by protein molecules, so that aggregation among micelles induced by calcium ions is weakened; the Tg enzyme crosslinking treatment and the citrate ions added, the Tg enzyme crosslinking treatment and the decalcification treatment and the Tg enzyme crosslinking treatment and the sodium ions added have synergistic effects, can better maintain the natural polymeric structure of the micelle, can effectively inhibit aggregation among the micelle in the high-temperature sterilization and subsequent storage of the concentrated solution, and ensures that the concentrated solution has higher thermal stability and lower viscosity.
The first object of the present invention is to provide a method for preparing a casein micelle concentrate with high thermal stability and low viscosity, which is any one of the following schemes (a), (b), (c), (d), (e), (f) and (g):
(a) Preparing MCC or MPC complex solution, and then performing Tg enzyme crosslinking treatment to obtain casein micelle concentrated solution;
(b) Preparing MCC or MPC complex solution, and then adding citrate ions to prepare casein micelle concentrated solution;
(c) Preparing MCC or MPC complex solution, and then decalcification treatment is carried out to obtain casein micelle concentrated solution;
(d) Preparing MCC or MPC complex solution, and then adding sodium ions to prepare casein micelle concentrated solution;
(e) Combining schemes (a) and (b) to produce a casein micelle concentrate; preparing MCC or MPC complex solution, then performing Tg enzyme crosslinking treatment, and adding citrate ions to prepare casein micelle concentrate;
(f) Combining schemes (a) and (c) to produce a casein micelle concentrate; preparing MCC or MPC complex solution, and then performing Tg enzyme crosslinking treatment and decalcification treatment to obtain casein micelle concentrated solution;
(g) Combining schemes (a) and (d) to produce a casein micelle concentrate; preparing MCC or MPC complex solution, then performing Tg enzyme crosslinking treatment and adding sodium ions to prepare the casein micelle concentrated solution.
In one embodiment of the present invention, the preparation method of the MCC comprises the following steps: taking pasteurized and defatted animal milk, carrying out microfiltration by adopting a ceramic membrane with the pore diameter of 50-200nm until the volume concentration multiple is 3-5, then adding water to the initial volume of the defatted milk, continuing to carry out washing filtration until the volume concentration multiple is 3-5, and repeating the washing filtration step for 2-4 times, wherein the microfiltration and washing filtration temperature are controlled to be 45-50 ℃ to obtain MCC trapped fluid; spray drying the trapped fluid, wherein the inlet air temperature and the outlet air temperature are respectively 130-140 ℃ and 70-80 ℃ to obtain MCC powder, and the dry protein content is 85-90%.
In one embodiment of the invention, the method for preparing the MPC comprises the following steps: taking pasteurized and defatted animal milk, ultrafiltering with organic membrane with molecular weight cut-off of 5-20kDa to volume concentration multiple of 3-5, adding water to the initial volume of defatted milk, continuing washing and filtering to volume concentration multiple of 3-5, repeating washing and filtering steps for 2-4 times, and controlling ultrafiltration and washing and filtering temperature at 45-50deg.C to obtain MPC retentate; spray drying the trapped fluid, wherein the inlet air temperature and the outlet air temperature are respectively 130-140 ℃ and 70-80 ℃, and the MPC powder is obtained, and the dry protein content is 85-90%.
In one embodiment of the invention, the animal milk comprises cow milk, goat milk, sheep milk, buffalo milk, yak milk, dromedary milk, bactrian camel milk, horse milk, donkey milk, and the like.
In one embodiment of the present invention, the method for preparing the complex solution includes the steps of: dissolving MCC or MPC in water, constant volume to casein content of 5-15% (w/v), dissolving at 200-400rpm for 20-40min, and homogenizing under 20-30mPa for 2-4 times.
In one embodiment of the invention, the method of Tg enzyme cross-linking treatment comprises the steps of: adding Tg enzyme at 200-400rpm until the ratio of enzyme activity to casein is 2-4U/g casein, maintaining at 35-45deg.C for 1-10 hr, and maintaining at 65-75deg.C for 1-3min to inactivate enzyme; wherein the enzyme activity of the Tg enzyme is 50-1000U/g.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 30% to 90%.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 30% to 50%.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 30% to 40%.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 40% to 50%.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 50% to 90%.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 50% to 70%.
In one embodiment of the invention, the Tg enzyme cross-linking treatment results in a casein micelle concentrate having a kappa-casein cross-linking of 70% to 90%.
In one embodiment of the invention, the citrate ion is added to a concentration of 8-22mM citrate ion per 100g casein.
In one embodiment of the invention, the citrate ion is added to a concentration of 8-16mM citrate ion per 100g casein.
In one embodiment of the invention, the citrate ion is added to a concentration of 8-12mM citrate ion per 100g casein.
In one embodiment of the invention, the decalcification treatment achieves a decalcification rate of 9% to 25%.
In one embodiment of the invention, the decalcification treatment achieves a decalcification rate of 9% to 20%.
In one embodiment of the invention, the decalcification treatment achieves a decalcification rate of 9% to 15%.
In one embodiment of the invention, the decalcification treatment achieves a decalcification rate of 9.5%.
In one embodiment of the invention, the sodium ions are added to a concentration of 160-500mM sodium ions per 100g casein.
In one embodiment of the invention, the sodium ions are added to a concentration of 160-350mM sodium ions per 100g casein.
In one embodiment of the invention, the citrate ion is carried by a mixture of trisodium citrate and citric acid; wherein, the mole ratio of trisodium citrate to citric acid in the mixture is 7.5:2.5-8.5:1.5, and the preferred mole ratio of trisodium citrate to citric acid is 8.0:2.0.
In one embodiment of the present invention, the method of decalcification treatment comprises the steps of: adding ion exchange resin Amberlite SR1L Na, maintaining at 200-400rpm for 2-4 hr, and filtering with 150-250 mesh filter bag to remove resin; wherein the amount of the ion exchange resin added is 9-25g resin/100 g casein.
In one embodiment of the invention, the sodium ions are carried in by sodium chloride.
In one embodiment of the present invention, the preferred (e), (f) and (g) schemes are followed by a Tg enzymatic crosslinking treatment.
The invention also provides casein micelle concentrated solution with high heat stability and low viscosity by using the method.
A second object of the present invention is to apply the above-mentioned high thermal stability and low viscosity casein micelle concentrate to the field of dairy processing.
A third object of the present invention is to apply the above-mentioned high thermal stability and low viscosity casein micelle concentrate to the field of protein concentrate product processing.
[ advantageous effects ]
The preparation method of the casein micelle concentrated solution with high thermal stability and low viscosity can better maintain the natural polymerization structure of the casein micelle, can effectively inhibit aggregation and precipitation or gelation among the casein micelle in high-temperature sterilization and subsequent storage of the concentrated solution, and ensures that the casein micelle concentrated solution has higher thermal stability, low relative viscosity and easy drinking. The invention combines Tg enzyme crosslinking treatment and adding citrate ions, combines Tg enzyme crosslinking treatment and decalcification treatment, combines Tg enzyme crosslinking treatment and adding sodium ions, and has synergistic effect in improving the heat stability of casein micelle concentrate and keeping the low viscosity sense of casein micelle concentrate.
Drawings
FIG. 1 is a flow chart of a preparation process of casein micelle concentrated solution in the invention.
FIG. 2 is an appearance of the casein micelle concentrate of example 2 after sterilization, storage and centrifugation.
FIG. 3 is an appearance of the casein micelle concentrate of example 4 after sterilization, storage and centrifugation.
Detailed Description
The following description of the preferred embodiments of the present invention is provided for better illustration of the invention, and should not be construed as limiting the invention.
The testing method comprises the following steps:
1. determination of protein content
The protein content was measured by the Kai-type nitrogen determination method, and the conversion coefficient was 6.38.
2. Determination of lactose content
Measurement of lactose content referring to national standard GB5413.5-2010, "measurement of lactose and sucrose in infant food and dairy products, national standard for food safety", an e2695 high performance liquid chromatograph (Waters corp., milford, MA, USA) was used, and a chromatographic column of XBridge Amide (250 mm×4.6 mm) was used, using a differential refractive index detector.
3. Determination of calcium content
The determination of the calcium content refers to national standard GB5009.268-2016 for determination of multiple elements in food safety national standard food, and inductively coupled plasma mass spectrometry is adopted.
4. Analysis of thermal stability
And (3) evaluating the thermal stability of the concentrated solution by adopting thermal coagulation time (HCT), loading 2mL of the concentrated solution into an ampoule bottle, heat-sealing, placing the ampoule bottle into an oil bath at 120 ℃, and recording the time from the ampoule bottle to flocculation, namely the HCT.
5. Determination of viscosity
The viscosity of the concentrated solutions is measured by adopting an Ubbelohde viscometer, and the viscosity ratio of each concentrated solution to the control concentrated solution is the relative viscosity.
6. Determination of the degree of crosslinking of kappa-Casein
Mixing the concentrated solution with Bis-tris buffer solution containing dithiothreitol, measuring kappa-casein content by using e2695 high performance liquid chromatograph, and detecting with XB ridge BEH C18 (250 mm×4.6 mm) with detection wavelength of 220nm.
7. Measurement of Casein micelle particle size
The particle size distribution of the casein micelles was determined using a Malvern nanosize potentiometer (Malvern Instruments ltd., malvern, worcestershire, UK), with the protein particle refractive index set at 1.57.
8. Determination of free Casein ratio
The concentrate was centrifuged at 150000g at 25℃for 1 hour, and the supernatant was collected. The casein content in the concentrate and supernatant was measured using an e2695 HPLC, using XB ridge BEH C18 (250 mm. Times.4.6 mm), with a detection wavelength of 220nm.
9. Measurement of the hydration Rate of Casein micelles
Taking concentrated solution, centrifuging at 150000g and 25 ℃ for 1h, taking precipitate, namely casein micelle, and drying at 103 ℃ for 7h, thereby measuring the dry basis moisture content of the precipitate, namely the hydration rate of the casein micelle.
EXAMPLE 1 preparation of MCC and MPC
The preparation steps of MCC are as follows: taking fresh goat milk, degreasing the fresh goat milk by adopting a disc type centrifuge at 9000rpm, and sterilizing the degreased milk (72 ℃ for 15 s); microfiltration is carried out on the skim milk by adopting a ceramic membrane with the aperture of 100nm until the volume concentration multiple is 4, then deionized water is added to the initial volume of the skim milk, washing and filtering are continued until the volume concentration multiple is 4, washing and filtering steps are repeated for 3 times, and the microfiltration and washing and filtering temperature are controlled to be 45-50 ℃ to obtain MCC trapped fluid; spray drying the trapped fluid, wherein the inlet air temperature and the outlet air temperature are 135 ℃ and 75 ℃ respectively, and the MCC powder is obtained.
The preparation steps of MPC are as follows: and (3) regulating microfiltration in the MCC preparation step to ultrafiltration, adopting an organic membrane with the molecular weight cutoff of 10kDa, and keeping other steps consistent with the MCC preparation step to obtain MPC trapped fluid and powder.
The dry basis compositions of the MCC and MPC prepared above are shown in Table 1.
TABLE 1 basic composition of MPC and MCC in example 1
Example 2 preparation of casein micelle concentrate by MCC: influence of Tg enzyme crosslinking treatment
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the total protein content is 6.1% (w/v), namely, the casein content is 6.0% (w/v), dissolving for 30min at a rotating speed of 300rpm, and then circularly homogenizing for 3 times under 25mPa to obtain MCC compound solution;
adding transglutaminase (Tg enzyme) into MCC compound solution at a speed of 300rpm until the enzyme activity concentration is 180U/L, keeping the ratio of enzyme activity to casein at 3U/g casein at 40 ℃ for 0, 1, 2, 4, 8 and 12 hours, and then keeping at 70 ℃ for 2 minutes to inactivate enzyme; wherein the Tg enzyme activity is 100U/g.
The physicochemical properties of the casein micelle concentrate prepared above are shown in table 2. As can be seen from the table, as Tg enzyme treatment time increases from 0h to 8h, the degree of crosslinking of kappa-casein increases gradually, the HCT of the concentrate also increases gradually, indicating that its thermal stability increases gradually, while the particle size of the casein micelles increases slowly, so that the relative viscosity of the concentrate also increases slowly; when the Tg enzyme treatment time is further increased to 12 hours, the crosslinking degree of kappa-casein is slightly increased, the particle size of casein micelle is greatly increased, the HCT of the concentrated solution is slightly reduced, and the relative viscosity of the concentrated solution is greatly increased. To analyze the storage stability of the concentrate, it was subjected to sterilization treatment (120 ℃ C., 20 s), then left at 4 ℃ C. For 14 days, and then centrifuged at 1000g for 5min. As can be seen from FIG. 2, when the degree of crosslinking of kappa-casein is 30.2% or more, no significant precipitate is formed after sterilization, storage and centrifugation of the concentrate. Tg enzyme treatment can crosslink kappa-casein on the surface of casein micelle, inhibit dissociation of kappa-casein from micelle surface during heat treatment, and inhibit calcium-sensitive casein (. Alpha.) in micelle s Exposing/beta), thereby inhibiting aggregation between casein micelles induced by calcium ions, thus improving the thermal stability of the casein micelle concentrate; when the Tg enzyme treatment time is too long, cross-linking is generated among casein micelles, and further, the heat stability of the casein micelle concentrate is reduced and the viscosity is increased. Therefore, in preparing a casein micelle concentrate by MCC, it is preferable that the degree of crosslinking of kappa-casein is 30.2 to 89.3%, so that the concentrateHas higher thermal stability and lower viscosity.
TABLE 2 physicochemical Properties of Casein micelle concentrate in example 2
Note that: the difference between the corresponding data is marked by the lower-case letters in the same column (P < 0.05)
Example 3 preparation of casein micelle concentrate by MCC: effect of mole ratio of trisodium citrate to citric acid
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding a mixture of trisodium citrate and citric acid to the MCC complex solution at 500rpm until the concentration of citrate ions is 10mM/L, corresponding to a ratio of citrate ions to casein of 16.7mM citrate ions/100 g casein; the molar ratio of trisodium citrate to citric acid in the mixture is 6.5:3.5, 7.0:3.0, 7.5:2.5, 8.0:2.0, 8.5:1.5, 9.0:1.0, or 9.5:0.5.
The physicochemical properties of the casein micelle concentrate prepared above are shown in table 3. As can be seen from the table, as the molar ratio of trisodium citrate to citric acid increases, the pH, HCT and free casein ratio of the concentrate all tended to increase, and the relative viscosity of the concentrate tended to decrease and then increase. When the mole ratio of trisodium citrate to citric acid is between 7.5:2.5 and 8.5:1.5, the pH of the concentrated solution is close to that of the concentrated solution without adding citrate ions, and the pH, HCT, free casein ratio and relative viscosity of the concentrated solution are less changed with the increase of the mole ratio of trisodium citrate to citric acid; when the proportion of trisodium citrate is too high, the pH of the concentrated solution is increased to be alkaline, the negative charge carried by casein is increased, dissociation of casein micelle is promoted, and the viscosity of the concentrated solution is increased; when the ratio of citric acid is too high, the pH of the concentrate is lowered to be slightly acidic, the negative charge of casein is reduced, and the binding between casein is promoted, resulting in an increase in the viscosity of the concentrate. Thus, in preparing a casein micelle concentrate by MCC, it may be preferable to have a molar ratio of trisodium citrate to citric acid of 7.5:2.5-8.5:1.5, to maintain the pH of the concentrate at neutral, and to provide the concentrate with higher thermal stability and lower viscosity.
TABLE 3 physicochemical Properties of Casein micelle concentrate in example 3
Note that: the difference between the corresponding data is significant (P < 0.05) as indicated by the difference between the lower-case letters in the same column.
Example 4 preparation of casein micelle concentrate by MCC: influence of citrate ion concentration
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding a mixture of trisodium citrate and citric acid to the MCC complex solution at 500rpm to a citrate ion concentration of 0, 2.5, 5.0, 7.5, 10.0, 12.5 or 15.0mM/L, corresponding to a citrate ion to casein ratio of 0, 4.2, 8.3, 12.5, 16.7, 20.8 or 25.0mM citrate ion per 100g casein; the molar ratio of trisodium citrate to citric acid in the mixture was 8.0:2.0.
The physicochemical properties of the casein micelle concentrate prepared above are shown in table 4. As can be seen from the table, the HCT of the concentrate gradually increased with increasing citrate ion concentration, indicating a gradual increase in its thermal stability. When the concentration of citrate ions is increased from 0mM/L to 12.5mM/L, the ratio of free casein is slowly increased, so that the relative viscosity of the concentrated solution is slowly increased; when the concentration of citrate ions was further increased to 15.0mM/L, the ratio of free casein was greatly increased, resulting in a great increase in the relative viscosity of the concentrate. To analyze the storage stability of the concentrate, it was subjected to sterilization treatment (120 ℃ C., 20 s), then left at 4 ℃ C. For 14 days, and then centrifuged at 1000g for 5min. As can be seen from FIG. 3, when the concentration of citrate ions was 5mM or more, no significant precipitate was formed after sterilization, storage and centrifugation. Citrate ions can chelate free calcium ions in whey, inhibit aggregation among casein micelles induced by the calcium ions, and therefore improve the thermal stability of casein micelle concentrate; the citrate ions also promote dissociation of calcium ions bound in the casein micelles into whey, which dissociates the casein micelle portions, and the dissociated casein molecules result in an increase in the viscosity of the concentrate. Thus, in preparing a casein micelle concentrate by MCC, it may be preferable that the citrate ion concentration be 5-12.5mM/L, i.e., 8.3-20.8mM citrate ion/100 g casein, to provide a concentrate with higher thermal stability and lower viscosity.
TABLE 4 physicochemical Properties of Casein micelle concentrate in example 4
Note that: the difference between the corresponding data is significant (P < 0.05) as indicated by the difference between the lower-case letters in the same column.
Example 5 preparation of casein micelle concentrate by MCC: influence of decalcification treatment
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding ion exchange resin Amberlite SR1L Na into the MCC complex solution, maintaining for 3 hours at a rotating speed of 300rpm, and filtering by adopting a 200-mesh filter bag to remove the resin; wherein the amount of ion exchange resin added is 0, 0.28, 0.56, 0.84, 1.12, 1.39 or 1.67g/100g of complex solution, corresponding to a ratio of ion exchange resin to casein of 0, 4.7, 9.3, 14.0, 18.7, 23.2 or 27.8g resin/100 g casein.
The physicochemical properties of the casein micelle concentrate prepared above are shown in table 5. As can be seen from the table, the HCT of the concentrate gradually increased with increasing decalcification, indicating a gradual increase in its thermal stability. When the decalcification rate is increased from 0% to 23.8%, the ratio of free casein is slowly increased, so that the relative viscosity of the concentrated solution is slowly increased; when the decalcification rate was further increased to 28.6%, the ratio of free casein was greatly increased, resulting in a great increase in the relative viscosity of the concentrate. The concentrate was subjected to sterilization treatment (120 ℃ C., 20 s) and then left at 4 ℃ C. For 14 days, followed by centrifugation at 1000g for 5 minutes, and it was found that no significant precipitate was formed in the concentrate when the decalcification rate was 9.5% or more. The removal of calcium ions in the concentrated solution can inhibit aggregation among casein micelles induced by the calcium ions, so that the heat stability of the casein micelle concentrated solution can be improved; the removal of calcium ions also causes dissociation of the casein micelle portion, and the dissociated casein molecules result in an increase in the viscosity of the concentrate. Therefore, when preparing casein micelle concentrates by MCC, it may be preferable to have a decalcification ratio of 9.5-23.8, resulting in a concentrate with higher thermal stability and lower viscosity.
TABLE 5 physicochemical Properties of Casein micelle concentrate in example 5
Note that: the difference between the corresponding data is significant (P < 0.05) as indicated by the difference between the lower-case letters in the same column.
Example 6 preparation of casein micelle concentrate by MCC: influence of sodium ion concentration
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
sodium chloride was added to the MCC multiplex solution at 500rpm to a sodium ion concentration of 0, 50, 100, 200, 300 or 400mM/L, corresponding to a sodium ion to casein ratio of 0, 83, 167, 333, 500 or 667mM sodium ion per 100g casein.
The physicochemical properties of the casein micelle concentrate prepared above are shown in table 6. As can be seen from the table, the HCT of the concentrate gradually increased as the concentration of sodium ions increased from 0mM/L to 300mM/L, indicating a gradual increase in thermal stability; when the concentration of sodium ions was further increased to 400mM/L, HCT of the concentrate was greatly reduced, indicating that its thermal stability was greatly reduced. As the concentration of sodium ions increases, the hydration rate of casein micelles slowly increases, so that the relative viscosity of the concentrate slowly increases. The concentrate was subjected to sterilization treatment (120 ℃ C., 20 s) and then left at 4 ℃ C. For 14 days, followed by centrifugation at 1000g for 5min, and it was found that no significant precipitate was formed in the concentrate when the concentration of sodium ions was between 100 and 400 mM. The sodium ions can increase the ionic strength of the concentrated solution, further increase the electrostatic shielding effect of negative charge groups in casein molecules, inhibit aggregation among casein micelles induced by calcium ions, and improve the thermal stability of the concentrated solution; when the electrostatic shielding effect of negative charge groups in casein molecules is overlarge, electrostatic repulsive force between protein molecules is too low, aggregation between casein micelles is promoted, and the thermal stability of the concentrate is reduced; sodium ions also increase the hydration rate of casein micelles, thereby increasing the viscosity of the concentrate. Thus, in preparing a casein micelle concentrate by MCC, it may be preferable that the concentration of sodium ions be 100-300mM/L, i.e., 167-500mM sodium ions per 100g casein, to provide a concentrate with higher thermal stability and lower viscosity.
TABLE 6 physicochemical Properties of Casein micelle concentrate in example 6
Note that: the difference between the corresponding data is significant (P < 0.05) as indicated by the difference between the lower-case letters in the same column.
Example 7 preparation of casein micelle concentrate by MCC: combination of Tg enzyme crosslinking treatment and addition of citrate ions
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding Tg enzyme into MCC complex solution at a rotating speed of 300rpm until the enzyme activity concentration is 180U/L, keeping the ratio of the corresponding enzyme activity to casein at 3U/g casein at 40 ℃ for 2 hours, and then keeping at 70 ℃ for 2 minutes to inactivate enzyme;
adding a mixture of trisodium citrate and citric acid to the MCC complex solution at 500rpm until the concentration of citrate ions is 5.0mM/L, corresponding to a ratio of citrate ions to casein of 8.3mM citrate ions per 100g casein; the molar ratio of trisodium citrate to citric acid in the mixture was 8.0:2.0.
Physicochemical properties of the casein micelle concentrates prepared in examples 7, 2 and 4 are shown in table 7. As can be seen from the table, the Tg enzyme cross-linking treatment alone increased the HCT of the concentrate by 15.4s compared to the concentrate without Tg enzyme cross-linking treatment and without citrate ion added; the citrate ions are added independently, so that the HCT of the concentrated solution is increased by 18.3s; the combination of the Tg enzyme crosslinking treatment and the addition of citrate ions increases the HCT of the concentrate by 45.4s, which is far greater than the effect of the Tg enzyme crosslinking treatment alone and the addition of citrate ions alone, namely 33.7s, which shows that the Tg enzyme crosslinking treatment and the addition of citrate ions have a synergistic effect in improving the heat stability of the casein micelle concentrate. Compared with the method that citrate ions are added independently, the combination of Tg enzyme crosslinking treatment and the addition of citrate ions reduces the ratio of free casein, and further reduces the relative viscosity of the concentrated solution. Therefore, when the casein micelle concentrated solution is prepared through MCC, tg enzyme crosslinking treatment and citrate ion addition are combined, and the Tg enzyme crosslinking treatment and the citrate ion addition are synergistic, so that the casein micelle concentrated solution has higher heat stability and lower viscosity.
TABLE 7 physicochemical Properties of Casein micelle concentrates in examples 7, 2 and 4
Note that: the difference between the corresponding data is marked by the lower-case letters in the same column (P < 0.05)
Example 8 preparation of casein micelle concentrate by MCC: adding citrate ion and Tg enzyme to combine with crosslinking treatment
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding a mixture of trisodium citrate and citric acid to the MCC complex solution at 500rpm until the concentration of citrate ions is 5.0mM/L, corresponding to a ratio of citrate ions to casein of 8.3mM citrate ions per 100g casein; the molar ratio of trisodium citrate to citric acid in the mixture is 8.0:2.0;
at 300rpm, tg enzyme was added to the MCC complex solution to an enzyme activity concentration of 180U/L, corresponding to a ratio of enzyme activity to casein of 3U/g casein, maintained at 40℃for 2 hours, and thereafter maintained at 70℃for 2 minutes to inactivate the enzyme.
Physicochemical properties of the casein micelle concentrates prepared in examples 7 and 8 are shown in table 8. As can be seen from the table, the Tg enzyme cross-linking treatment followed by the citrate ion added resulted in a longer HCT of the concentrate and a lower free casein ratio and thus a lower relative viscosity of the concentrate than the Tg enzyme cross-linking treatment followed by the citrate ion added. Therefore, when preparing the casein micelle concentrate through MCC, tg enzyme crosslinking treatment and adding citrate ions can be combined, and the Tg enzyme crosslinking treatment is preferable, and then the citrate ions are added, so that the casein micelle concentrate has higher thermal stability and lower viscosity.
Table 8 physicochemical Properties of casein micelle concentrates in examples 7 and 8
Note that: the difference between the corresponding data is marked by the lower-case letters in the same column (P < 0.05)
Example 9 preparation of casein micelle concentrate by MCC: combination of Tg enzyme crosslinking treatment and decalcification treatment
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding Tg enzyme into MCC complex solution at a rotating speed of 300rpm until the enzyme activity concentration is 180U/L, keeping the ratio of the corresponding enzyme activity to casein at 3U/g casein at 40 ℃ for 2 hours, and then keeping at 70 ℃ for 2 minutes to inactivate enzyme;
adding ion exchange resin Amberlite SR1L Na, maintaining at 300rpm for 3h, and filtering with 200 mesh filter bag to remove resin; wherein the amount of the ion exchange resin added was 0.56g/100g of the complex solution, and the ratio of the ion exchange resin to casein was 9.3g of the resin/100 g of casein.
Physicochemical properties of the casein micelle concentrates prepared in examples 9, 2 and 5 are shown in table 9. As can be seen from the table, the Tg enzyme cross-linking treatment alone increased the HCT of the concentrate by 15.4s compared to the concentrate without Tg enzyme cross-linking treatment and without decalcification treatment; the HCT of the concentrated solution is increased by 19.3s by single decalcification treatment; the combination of the Tg enzyme cross-linking treatment and the decalcification treatment increases the HCT of the concentrate by 44.8s, which is much greater than the effect of the Tg enzyme cross-linking treatment and decalcification treatment alone, i.e., 34.7s, indicating that the Tg enzyme cross-linking treatment and decalcification treatment have a synergistic effect in improving the thermal stability of the casein micelle concentrate. The combination of Tg enzyme cross-linking and decalcification reduces the free casein ratio compared to decalcification alone, which in turn reduces the relative viscosity of the concentrate. Therefore, when the casein micelle concentrate is prepared through MCC, tg enzyme crosslinking treatment and decalcification treatment can be combined, and the Tg enzyme crosslinking treatment and decalcification treatment are synergistic, so that the casein micelle concentrate has higher thermal stability and lower viscosity.
Table 9 physicochemical Properties of casein micelle concentrates in examples 9, 2 and 5
Note that: the difference between the corresponding data is marked by the lower-case letters in the same column (P < 0.05)
Example 10 preparation of casein micelle concentrate by MCC: decalcification treatment and Tg enzyme crosslinking treatment are combined
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding ion exchange resin Amberlite SR1L Na, maintaining at 300rpm for 3h, and filtering with 200 mesh filter bag to remove resin; wherein the addition amount of the ion exchange resin is 0.56g/100g of complex solution, and the ratio of the corresponding ion exchange resin to casein is 9.3g of resin/100 g of casein;
at 300rpm, tg enzyme was added to the MCC complex solution to an enzyme activity concentration of 180U/L, corresponding to a ratio of enzyme activity to casein of 3U/g casein, maintained at 40℃for 2 hours, and thereafter maintained at 70℃for 2 minutes to inactivate the enzyme.
Physicochemical properties of the casein micelle concentrates prepared in examples 9 and 10 are shown in table 10. As can be seen from the table, the Tg enzyme cross-linking treatment followed by the decalcification treatment resulted in a longer HCT of the concentrate and a lower free casein ratio and thus a lower relative viscosity of the concentrate than the decalcification treatment followed by the Tg enzyme cross-linking treatment. Thus, when preparing the casein micelle concentrate by MCC, the combination of Tg enzyme crosslinking treatment and decalcification treatment is adopted, and the Tg enzyme crosslinking treatment and decalcification treatment are preferable, so that the casein micelle concentrate has higher thermal stability and lower viscosity.
Table 10 physicochemical Properties of casein micelle concentrates in examples 9 and 10
Note that: the difference between the corresponding data is marked by the lower-case letters in the same column (P < 0.05)
Example 11 preparation of casein micelle concentrate by MCC: combination of Tg enzyme crosslinking treatment and sodium ion addition
The preparation of the casein micelle concentrated solution comprises the following steps:
dissolving MCC powder prepared in example 1 in deionized water to a constant volume until the casein content is 6.0% (w/v), dissolving for 30min at 300rpm, and then circularly homogenizing for 3 times at 25mPa to obtain MCC compound solution;
adding Tg enzyme into MCC complex solution at a rotating speed of 300rpm until the enzyme activity concentration is 180U/L, keeping the ratio of the corresponding enzyme activity to casein at 3U/g casein at 40 ℃ for 2 hours, and then keeping at 70 ℃ for 2 minutes to inactivate enzyme;
sodium chloride was added to the MCC multiplex solution at 500rpm to a sodium ion concentration of 100mM/L, corresponding to a sodium ion to casein ratio of 167mM sodium ion per 100g casein.
Physicochemical properties of the casein micelle concentrates prepared in examples 11, 2 and 6 are shown in table 11. As can be seen from the table, the Tg enzyme cross-linking treatment alone increased the HCT of the concentrate by 15.4s compared to the concentrate without Tg enzyme cross-linking treatment and without sodium ions added; sodium ions are added independently, so that HCT of the concentrated solution is increased by 15.2s; the combination of the Tg enzyme crosslinking treatment and the addition of sodium ions increases the HCT of the concentrate by 36.9s, which is greater than the effect of the Tg enzyme crosslinking treatment alone and the sodium ions alone, namely 30.6s, which indicates that the Tg enzyme crosslinking treatment and the sodium ions addition have a synergistic effect in improving the heat stability of the casein micelle concentrate. Compared with the method that sodium ions are added singly, the combination of Tg enzyme crosslinking treatment and sodium ions is used for reducing the hydration rate of casein micelles, and further reducing the relative viscosity of the concentrated solution. Therefore, when preparing the casein micelle concentrated solution through MCC, the combination of Tg enzyme crosslinking treatment and adding sodium ions can be adopted, and the Tg enzyme crosslinking treatment and the sodium ions are synergistic, so that the casein micelle concentrated solution has higher thermal stability and lower viscosity.
Table 11 physicochemical Properties of casein micelle concentrates in examples 11, 2 and 6
Note that: the difference between the corresponding data is marked by the lower-case letters in the same column (P < 0.05)
Example 12 preparation of casein micelle concentrate by MPC: influence of citrate ion concentration
The preparation of the casein micelle concentrated solution comprises the following steps:
taking MPC powder prepared in the example 1, redissolving the MPC powder in deionized water, fixing the volume until the total protein content is 7.7% (w/v), namely the casein content is 6.0% (w/v), dissolving the MPC powder for 30min at a rotating speed of 300rpm, and then circularly homogenizing the MPC powder for 3 times at 25mPa to obtain MPC complex solution;
adding a mixture of trisodium citrate and citric acid to the MPC complex solution at 500rpm to a citrate ion concentration of 0, 2.5, 5.0, 7.5, 10.0, 12.5 or 15.0mM/L, corresponding to a citrate ion to casein ratio of 0, 4.2, 8.3, 12.5, 16.7, 20.8 or 25.0mM citrate ion/100 g casein; the molar ratio of trisodium citrate to citric acid in the mixture was 8.0:2.0.
The physicochemical properties of the casein micelle concentrate prepared above are shown in table 12. As can be seen from the table, the HCT of the concentrate gradually increased with increasing citrate ion concentration, indicating a gradual increase in its thermal stability. When the concentration of citrate ions is increased from 0mM/L to 12.5mM/L, the ratio of free casein is slowly increased, so that the relative viscosity of the concentrated solution is slowly increased; when the concentration of citrate ions was further increased to 15.0mM/L, the ratio of free casein was greatly increased, resulting in a great increase in the relative viscosity of the concentrate. To analyze the storage stability of the concentrate, it was subjected to sterilization treatment (120 ℃ C., 20 s), then left at 4 ℃ C., for 14 days, and then centrifuged at 1000g for 5min, and it was found that no significant precipitate was formed in the concentrate when the concentration of citrate ions was 5mM or more. Thus, in preparing casein micelle concentrates by MPC, it may be preferable that the citrate ion concentration be in the range of 5 to 12.5mM/L, i.e., 8.3 to 20.8mM citrate ion per 100g casein, to provide the concentrate with higher thermal stability and lower viscosity.
TABLE 12 physicochemical Properties of Casein micelle concentrate in example 12
Note that: the difference between the corresponding data is significant (P < 0.05) as indicated by the difference between the lower-case letters in the same column.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (4)
1. A preparation method of casein micelle concentrated solution with high thermal stability and low viscosity is characterized in that,
preparing concentrated micelle casein or concentrated milk protein complex solution, performing Tg enzyme crosslinking treatment, and adding citrate ions to obtain casein micelle concentrated solution;
the preparation method of the complex solution comprises the following steps: redissolving the concentrated micelle casein or the concentrated milk protein in water, fixing the volume until the mass volume ratio content of the casein is 5-15%, dissolving for 20-40min at the rotating speed of 200-400rpm, and then circularly homogenizing for 2-4 times under the condition of 20-30 mPa; the protein dry basis content of the concentrated micelle-state casein or the concentrated milk protein is 85-90% and is derived from goat milk;
the Tg enzyme crosslinking treatment leads the kappa-casein crosslinking degree of the casein micelle concentrated solution to be 30% -90%; the ratio of Tg enzyme activity to casein is 2-4U/g casein;
the concentration of the added citrate ions is 8-22mM citrate ions/100 g casein;
the citrate ions are carried in by a mixture of trisodium citrate and citric acid; wherein, the mole ratio of trisodium citrate to citric acid in the mixture is 7.5:2.5-8.5:1.5;
the pH of the casein micelle concentrated solution is neutral.
2. The method of claim 1, wherein Tg enzyme cross-linking treatment is performed first.
3. A casein micelle concentrate prepared by the method of claim 1 or 2.
4. Use of the casein micelle concentrate of claim 3 in the field of dairy processing.
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| CN1832687A (en) * | 2003-08-07 | 2006-09-13 | 方塔拉合作集团有限公司 | Production of milk protein ingredient with high whey protein content |
| CN106455640A (en) * | 2014-04-11 | 2017-02-22 | N·V·努特里奇亚 | Stabilized micellar casein and compositions |
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| CN1832687A (en) * | 2003-08-07 | 2006-09-13 | 方塔拉合作集团有限公司 | Production of milk protein ingredient with high whey protein content |
| CN106455640A (en) * | 2014-04-11 | 2017-02-22 | N·V·努特里奇亚 | Stabilized micellar casein and compositions |
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