CN100358916C - Method for extracting high purity protein from cow milk or soybean waste water - Google Patents
Method for extracting high purity protein from cow milk or soybean waste water Download PDFInfo
- Publication number
- CN100358916C CN100358916C CNB2005100952443A CN200510095244A CN100358916C CN 100358916 C CN100358916 C CN 100358916C CN B2005100952443 A CNB2005100952443 A CN B2005100952443A CN 200510095244 A CN200510095244 A CN 200510095244A CN 100358916 C CN100358916 C CN 100358916C
- Authority
- CN
- China
- Prior art keywords
- resin
- milk
- lactoferrin
- desorbs
- bovinelactoperoxidase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 40
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 20
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 20
- 235000010469 Glycine max Nutrition 0.000 title 1
- 244000068988 Glycine max Species 0.000 title 1
- 235000020247 cow milk Nutrition 0.000 title 1
- 239000002351 wastewater Substances 0.000 title 1
- 239000011347 resin Substances 0.000 claims abstract description 64
- 229920005989 resin Polymers 0.000 claims abstract description 64
- 235000013336 milk Nutrition 0.000 claims abstract description 42
- 239000008267 milk Substances 0.000 claims abstract description 42
- 210000004080 milk Anatomy 0.000 claims abstract description 42
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 31
- 239000007788 liquid Substances 0.000 claims abstract description 29
- 239000012528 membrane Substances 0.000 claims abstract description 26
- 239000005862 Whey Substances 0.000 claims abstract description 16
- 102000007544 Whey Proteins Human genes 0.000 claims abstract description 16
- 108010046377 Whey Proteins Proteins 0.000 claims abstract description 16
- 239000000203 mixture Substances 0.000 claims abstract description 12
- 238000000926 separation method Methods 0.000 claims abstract description 7
- 238000010521 absorption reaction Methods 0.000 claims description 32
- 102000010445 Lactoferrin Human genes 0.000 claims description 23
- 108010063045 Lactoferrin Proteins 0.000 claims description 23
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 claims description 23
- 229940078795 lactoferrin Drugs 0.000 claims description 23
- 235000021242 lactoferrin Nutrition 0.000 claims description 23
- 101001099470 Bos taurus Lactoperoxidase Proteins 0.000 claims description 22
- 235000018102 proteins Nutrition 0.000 claims description 18
- 150000003839 salts Chemical class 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 17
- 239000000843 powder Substances 0.000 claims description 15
- 238000000108 ultra-filtration Methods 0.000 claims description 15
- 238000010612 desalination reaction Methods 0.000 claims description 14
- 238000001471 micro-filtration Methods 0.000 claims description 14
- 238000000967 suction filtration Methods 0.000 claims description 10
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 238000001179 sorption measurement Methods 0.000 claims description 6
- 238000005516 engineering process Methods 0.000 claims description 5
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 4
- 230000008929 regeneration Effects 0.000 claims description 4
- 238000011069 regeneration method Methods 0.000 claims description 4
- 238000001223 reverse osmosis Methods 0.000 claims description 4
- 159000000007 calcium salts Chemical class 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 239000001103 potassium chloride Substances 0.000 claims description 3
- 235000011164 potassium chloride Nutrition 0.000 claims description 3
- 159000000000 sodium salts Chemical group 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 2
- 238000002592 echocardiography Methods 0.000 claims description 2
- 238000000605 extraction Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 210000000481 breast Anatomy 0.000 claims 1
- 238000009776 industrial production Methods 0.000 abstract description 3
- 102000004190 Enzymes Human genes 0.000 abstract 3
- 108090000790 Enzymes Proteins 0.000 abstract 3
- 230000001590 oxidative effect Effects 0.000 abstract 3
- 239000003729 cation exchange resin Substances 0.000 abstract 1
- 229940023913 cation exchange resins Drugs 0.000 abstract 1
- 230000002708 enhancing effect Effects 0.000 abstract 1
- 238000001914 filtration Methods 0.000 abstract 1
- 101000798100 Bos taurus Lactotransferrin Proteins 0.000 description 43
- 229940072440 bovine lactoferrin Drugs 0.000 description 43
- 238000001035 drying Methods 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- VIKNJXKGJWUCNN-XGXHKTLJSA-N norethisterone Chemical compound O=C1CC[C@@H]2[C@H]3CC[C@](C)([C@](CC4)(O)C#C)[C@@H]4[C@@H]3CCC2=C1 VIKNJXKGJWUCNN-XGXHKTLJSA-N 0.000 description 6
- 239000012266 salt solution Substances 0.000 description 6
- 239000000706 filtrate Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 230000003115 biocidal effect Effects 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 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 3
- 241000894006 Bacteria Species 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 3
- 239000012267 brine Substances 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 239000003456 ion exchange resin Substances 0.000 description 3
- 229920003303 ion-exchange polymer Polymers 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 235000020183 skimmed milk Nutrition 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 3
- 241001635574 Sabatia angularis Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- -1 iron ion Chemical class 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 230000000452 restraining effect Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 235000008939 whole milk Nutrition 0.000 description 2
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- CWHJIJJSDGEHNS-MYLFLSLOSA-N Senegenin Chemical compound C1[C@H](O)[C@H](O)[C@@](C)(C(O)=O)[C@@H]2CC[C@@]3(C)C(CC[C@]4(CCC(C[C@H]44)(C)C)C(O)=O)=C4[C@@H](CCl)C[C@@H]3[C@]21C CWHJIJJSDGEHNS-MYLFLSLOSA-N 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000931197 Themeda Species 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 210000003022 colostrum Anatomy 0.000 description 1
- 235000021277 colostrum Nutrition 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000011169 microbiological contamination Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000037452 priming Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 210000003079 salivary gland Anatomy 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 235000020161 semi-skimmed milk Nutrition 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000009871 tenuigenin Substances 0.000 description 1
- 241000712461 unidentified influenza virus Species 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Landscapes
- Dairy Products (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention discloses a method for extracting high purity protein from milk or whey water. An agitating tank where cation exchange resins are placed is used for adsorbing, desorbing, separating and extracting lactoferritin, milk oxidative enzyme protein and mixtures of the lactoferritin and the milk oxidative enzyme protein in milk or whey liquid, and a membrane separation technique is used for separating, concentrating and highly purifying the lactoferritin, the milk oxidative enzyme protein and the mixtures. The rotary speed of agitating pulp in the agitating tank can be adjusted to be from 20 to 100 rpm; the side wall in the tank is provided with a swirl enhancing baffle plate, and the tank bottom is provided with a resin bed with a filter plate; a high-speed flow-guiding sucking and filtering pipe is arranged in a position of 8 to 12cm higher above a resin layer on the resin bed. The problem that industrial production is carried out by using low cost is solved.
Description
Technical field
The present invention relates to a kind of method of from milk or whey water, extracting high purity protein
Background technology
The present invention relates to a kind ofly from milk or whey water, extract highly purified lactoferrin (bovine lactoferrin simultaneously with stirred pot, be abbreviated as BLF), the method for Bovinelactoperoxidase (bovine lactoperoxidase is abbreviated as BLP) and lactoferrin and Bovinelactoperoxidase mixture.Bovinelactoperoxidase and lactoferrin have the antibiotic of wide spectrum, and be antiviral and improve the immunologic function of body disease-resistant.For example BLP can destroy the tenuigenin of germ, suppresses Yeast Nucleic Acid synthetic in the germ process of growth.Thereby it is to gram-positive microorganism and Gram-negative bacteria, yeast such as Candida albicans, and mould such as black-koji mould all have restraining effect.There is Bovinelactoperoxidase in the secretory product in the mammal, particularly in the milk.But its concentration is very low in first Ruzhong, and its concentration increases sharply and passes to the antibiotic of neonatal as parent after 4-5 days.Because the immune yarn system of neonatal is not physically well developed as yet, need from breast milk, be protected with germ and the virus that resists an invasion.
Bovinelactoferrin (BLF) is a ferruginous glycoprotein, and is opposite with Bovinelactoperoxidase, and it is dense colostrum, and its concentration just descends rapidly after several days.BLF also is a very antibacterial protein of wide spectrum.It can be robbed iron ion from germ, thereby germ mistake iron is suppressed or death.So BLF is to gram-positive microorganism such as streptococcus aureus, suis etc.; Gram-negative bacteria such as intestinal bacteria, dysentery bar mattress, and shiga bacillus; Hp, influenza virus, eczema virus and salivary gland virus all have restraining effect.
Because simple BLP and BLF have the proteic function of biological antibiotic, their two kinds of mixtures together also should have antibiotic, the function of antiviral and enhance immunity power.
The laboratory scale of existing report is isolated BLF, as uses DEAE, SephadexC50, CMSephaseFF, CMToyopear l, BioradMacro-preCM, Dowx-mac-3 and PuriliteC115 ion exchange resin.But these methods are resin to be adorned post use and adopt damping fluid.Such method is inapplicable large-scale industrial practice.Because BLF and the concentration of BLP in milk are very low, produce that commercial economy is worth, highly purified BLP and BLF must adopt the milk of huge amount so that enough BLF and BLP to be provided, such as will produce 20 kilograms of purity daily is 96% above BLF, suppose that its total recovery is 80%, the BLF in the raw material milk is 180mg/L.This just needs 160 tons of milk.And the treatment time of these 160 tons of milk must in 10 hours, finish, not only can increase the chance of milk microbiological contamination because consider long process period, and will set apart to ion exchange resin column regeneration and disinfecting to prepare second day work.Moreover, cause the absorption dynamics of BLF and BLP very slow because milk intermediate ion intensity is about 6-7mS (milli west door), needed 50 minutes just can reach more than 60% of total amount approximately.Therefore must give resin and milk has enough duration of contact.In order to make BLP as much as possible in the milk, BLF contacts the production permutoid reaction with resins exchange functional group simultaneously, must play the linear velocity that guarantees certain milk flowing, and in other words, linear velocity is big more in the post, and its resin absorption dynamics is just fast more.Such as the linear velocity in the post at least will 100cm per hour more than (1.6cm per minute), concerning the garden post that a diameter is 50cm, reach the linear velocity of 1.6cm per minute, its volumetric flow rate just at least will be at 3.2 Liter Per Minutes.If be no less than 50 minutes duration of contact, the length of post will be more than 83cm.But its column length surpasses 50cm when the diameter of post surpasses 30cm, just is difficult to guarantee the quality of its flow and dress post.Thereby above-mentioned these ion exchange resin are for laboratory scale designs, and do not possess the performance that still keeps high flow capacity under the technical scale condition, therefore can't be applied to the production process of scale like this.If moreover in separating the process of telling, adopt damping fluid, also increase production cost greatly, product can't be competed, this is worthless on industrial production especially.
Also have report to adopt ultra-filtration membrane to come from whey, to separate BLF, but the purity of the BLF that this method is produced is very low, only has an appointment about 30%.One of them possible factor is exactly that the molecular weight of BLP and BLF is very close, and the ultrafiltration membrane technique of present stage is impossible with these two kinds of albumen separately.
Have report to extract BLF, but because the iso-electric point of BLF and BLP is very close, this method can't be produced highly purified BLP and BLF with the isoelectric precipitation method.
Have report to use heparin agar affinity chromatography to extract BLF, but the production cost of this method is too high, can't realize technical scale.
Summary of the invention
Can not low cost carry out big industrial problem for solving the existing high purity protein technology of from milk or whey water, extracting.The invention provides a kind of method of from milk or whey water, extracting highly purified lactoferrin, Bovinelactoperoxidase and lactoferrin and Bovinelactoperoxidase mixture with stirred pot simultaneously.
Technical scheme of the present invention is as follows:
A kind of method of from milk or whey water, extracting high purity protein, the stirred pot that its adopt to place Zeo-karb adsorbs, desorbs, lactoferrin, newborn Bovinelactoperoxidase and both mixtures thereof in separation and Extraction milk or the whey liquid, and is aided with membrane separation technique and it is separated concentrates and highly purified.
Stirred pot is adjustable, and the stirring arm rotating speed is 20-100rpm; The jar inner side-wall has the eddy current of increasing baffle plate, and jar end is provided with the resin bed of band screen plate, and 8-12cm highly locates to be provided with quick water conservancy diversion suction filtration pipe above the resin layer on the resin bed.The various transmitter pH meters of controlling for automatically are set in jar, the weight meter, electric conductivity is taken into account thermometer, and the pot liquid holding temperature is 3-160 ℃.Resin is the SPC70 resin, and number of times that resin absorption desorbs and condition can be set according to the requirement of variant production and product purity; The adsorption time of resin is 20-180 minute, and adsorption temp is at 3-30 ℃.The adsorption time of resin is 35-60 minute.Desorbing of resin adopts the two-stage salt concn to desorb lactoferrin, newborn Bovinelactoperoxidase.First step salt concn is 1-2.5%, desorbs newborn Bovinelactoperoxidase, and the time of desorbing is 30-100 minute; Second stage salt concn is 3.5-7%, desorbs lactoferrin, and the time of desorbing is 30-100 minute; The salt concn that desorbs newborn Bovinelactoperoxidase and lactoferrin mixture is to be 30-100 minute the 3-7% time of desorbing; The stir speed (S.S.) that desorbs is 20-100rpm.It is sodium salt, calcium salt that resin desorbs used salt, sylvite; Resin need not regeneration activating after absorption and desorbing technological process, can be for the usefulness of next batch absorption as long as be lower than 1.6ms with reverse osmosis water (RO water) rinsing resin to its ionic strength after desorbing the technology end.Membrane separation technique uses the microfiltration membrane of 0.05-0.1 micron with lactoferrin, newborn Bovinelactoperoxidase separate the attached liquid degerming, remove remaining fat and other small-particle; Ultra-filtration membrane with 1K-30K molecular interception amount carries out desalination and concentrated.Highly purified lactoferrin is that the lactoferrin dope that will desorb desalination and concentration through the absorption of above-mentioned technology is separated the concentrated acquisition that echos ultra-filtration membrane through the absorption of weak-type sun resin such as CM resin again.With adsorb, desorb, the lactoferrin dope of desalination and concentration, newborn Bovinelactoperoxidase dope and lactoferrin thereof, newborn Bovinelactoperoxidase mixing dope carries out vacuum dehydrating at lower temperature or low temperature spray drying becomes powder mixture.
Adopt the form of stirred pot rather than dress post to carry out absorption and separate the process of telling.Thereby need not adorn post and worry the resin column problem of short-circuit.Can increase stirring velocity and separate the kinetics speed of telling to increase absorption.In stirred pot, increase baffle plate, thereby make proteic transmission speed faster.In addition-the more important thing is that installation-suction filtration tube is accelerated drain age velocity on the resin bed surface in jar, in just big industrial production.
Description of drawings
Fig. 1 is a process flow diagram of the present invention
Embodiment
1, adopt the form of stirred pot rather than dress post to carry out absorption and separate the process of telling.Thereby need not adorn post and worry the resin column problem of short-circuit.Can increase stirring velocity and separate the kinetics speed of telling to increase absorption.In stirred pot, increase baffle plate, thereby make proteic transmission speed faster.In addition-the more important thing is that (dotted line among Fig. 1 in the T2 jar) installation-suction filtration tube (FT among Fig. 1 in the T2 jar) is accelerated drain age velocity on the resin bed surface in jar.This is because when absorption and separate when telling to finish, and stirring arm stops stirring.Resin (the thick black line of the level among Fig. 1 in the T2 jar) on the screen plate at the bottom of the action of gravity deposit jar forms resin bed.If at this moment too thin the or too soft liquid that will cause of resin particle is difficult to by resin bed, thereby the flow of discharge opeing diminishes at the bottom of causing jar, and the activity duration is long, and productive rate descends.In order to accelerate drain age velocity, installation-suction filtration pipe on the resin bed surface, make most liquid without resin bed directly through suction filtration pipe and valve V25, V5, VM1 and V6 discharge.When the liquid level in the jar dropped to the resin bed surface, suction filtration pipe FT stopped drawing liquid, remaining liq V5 at the bottom of jar in jar, and VM1 and V6 extract out.Such process is just improved the speed of discharge opeing greatly.
2, this law does not need raw material milk or whey water are regulated pH and ionic strength.And utilize the natural pH and the ionic strength of milk or whey water.Its normal ph should be 6.2-6.9
3, the milk of this paper middle finger is meant heating or the not new fresh milk of thermal sterilization or stale milk and the milk that is mixed with milk powder.
4, this method can directly adopt whole milk and skimmed milk.Or the full-cream or skimmed milk of reduction that is mixed with milk powder.
5, this method can directly be used whey water.
6, Zeo-karb all can be used as the BLF of this method and the absorption resin of BLP, but SPC70 is the first-selected absorption resin of this law.
7, the 20-120 minute absorption time of this paper employing, the absorption time is too short, and yield is low excessively, and the absorption time is long, can improve yield, considers long process period but play.Absorption and separate tell temperature can be at 3-16 degree C.
8, adopt two-stage salt (sodium-chlor) concentration to separate and tell BLP and BLF.First step salt concn is that 1-2.5% separates and tells BLP, and separating the time of telling is 30-100 minute.Second stage salt concn is that 3.5-7% separates and tells BLF, and separating the time of telling is 30-100 minute.
9, this law used salt in separating and telling can be sodium salt such as sodium-chlor etc., can be calcium salt such as calcium chloride etc., also can be sylvite such as Repone K etc.But the first-selected sodium-chlor of this paper.
But 10, this law continuous production, promptly resin does not need other regeneration, and a dust is washed resin to its ionic strength with reverse osmosis water (RO water) and is lower than the absorption that 1.6mS can begin next batch after separating and telling to finish.
11, the stirring velocity in the stirred pot is at 30-70rpm.
12, adopt the microfiltration membrane degerming and the remaining fat of 0.05-0.1 micron, molecule etc.
13, the elutriant of BLP of Shou Jiing and BLP (separate and tell liquid) carries out desalination and concentrated with the ultra-filtration membrane of 1K-30K molecular retention amount.
14, as will producing highly purified BLF, desalination and concentrate good BLF again through the absorption (the T7 jar among Fig. 1) of the positive resin such as the CM resin of a weak-type to remove wherein a small amount of other foreign protein.
15, BLP after concentrating and the mixed solution of BLF, BLP solution and BLF solution can be used the also available spraying drying of freeze-drying drying (FD among Fig. 1) (SD among Fig. 1).Final protein powder packing (the square case among Fig. 1).
16, the leaching salt solution and can use reverse osmosis unit (RO) or electric osmose device for folding recovered brine of ultra-filtration membrane, the RO filtrate can be made water of productive use simultaneously.
T1 among Fig. 1 is milk container or whey water pot.After raw material is transported to stirred pot T2 from the T1 jar, stirred 60-120 minute.After absorption finishes, stop to stir and allowed in 3 minutes after the resin free setting, again by suction filtration villous themeda FT, VM1, V5, V6 is transported to the T3 jar with the milk liquid of absorption.This milk liquid still contains certain density BLP and BLF, so still can be considered as milk or other milk preparation usefulness.
Through the V3 valve with RO water wash resin (full-cream in this way or half fat milk, with the RO water wash of 34 degree C) more than 3 times, to remove milk liquid and the fat of remnants.Leacheate is discharged through valve V4.
(1) produces BLP and BLF mixed protein;
After adding 5-6% salt solution from the V2 valve, stir after 60-100 minute, stop to stir and allowed the resin natural subsidence in 3 minutes, again through FT, V25, V5 and VM5 will separate and tell liquid and be transported to the TMF jar.
To separate the particulate and the fat of telling in the liquid through 0.1 micron microfiltration membrane removes.Mocromembrane filtrate is transported to the T4 jar through VM2 and V8 again.Again from the T4 jar through V18, V15 is by the ultra-filtration membrane desalination and be concentrated to protein concentration more than 6%.Wherein V13 and V8 are the return lines of ultra-filtration membrane, and V7 is the priming valve door.
The concentrated solution of BLPF is transported to the spray-dryer drying or gets the dry powder finished product after V20 is transported to the lyophilizer drying through V21.Powder is light brown green.
Add RO water and be lower than 1.6 milli west doors at the resin that stirred pot T2 has separated after telling until ionic strength through the drip washing of V3 valve.Ring waste is discharged from the V4 valve.Stirred pot T2 now waits upon life and prepares the next round absorption.
(2) produce BLP monomer and BLF monomer;
First step salt hydrolysis is told BLP; Add 1.6-2.5% salt solution (sodium chloride brine) from the V2 valve, stirred 40-80 minute, stop to stir and began suction filtration in 3 minutes and separate and tell and through V25, V5, VM5 are transported to the TMF jar, and then through V2 1.6-2.5% salt water wash resin 3 times, leacheate is through V25, and V5, VM5 are discharged to the TMF jar.
Second stage salt hydrolysis is told BLF; Separate the resin adding 4-6% salt solution (sodium chloride brine) of telling from the V2 valve to finishing the first step, stirred 60-120 minute.
The second stage separate tell in, the microfiltration membrane unit is also told liquid to separating of the first step simultaneously and is filtered.Filtrate is through VM2, and V10 is transported to T5.After filter finishing, clean the microfiltration membrane unit and separate and tell liquid to prepare to accept the second stage.
Separate after the time of telling finishes the second stage, BLF is separated tells liquid through V25, after V5, VM5 are drained into the TMF jar, by the V3 valve add RO water wash resin more than 3 times to ionic strength be below the 1.6 milli west doors.Leacheate merges to the TMF jar.Stirred pot T2 recovered to wait upon life state Huaihe River get a collection of production ready the sixth of the twelve Earthly Branches this moment.
Starting the micro-filtration unit filters BLF in the TMF jar and separates and tell liquid.Filtered liquid is through VM2, and V12 is transported to the T6 jar.
Separate at the BLP of T5 and T6 and to tell liquid and BLF and separate and tell liquid successively by ultra-filtration membrane unit desalination and concentration.BLP is transported to drying unit through V17 then.Dried powder is a brown-green.
Then play the resin cation (R.C.) of process in the T7 jar to remove remaining impure milk proem to producing highly purified BLF.The desalting soln of BLF is through V16, and V23 is transported to the T7 jar, stirs absorption after 20-50 minute, and suction filtration liquid is transported to drying unit through V24 and carries out drying.Dry back powder is rose pink.
Embodiment one: 993 liters of semi-skimmed milks, and temperature 8.2 degree C, pH6.6, total protein are every liter of 32.5 gram, 366 milligrams every liter of BLP concentration, BLF concentration are 138 milliliters every liter.Adopt the operational path of Fig. 1, and produce the egg mix white powder of BLP and BLF by the described step of explanation.Dress SPEC70 resin in the stirred pot T2.115 minutes absorption time.Separate and add 90 liters of 5.5% sodium chloride brines when telling, separate the 65 minutes time of telling.Separate and tell and leacheate merges to micro-filtration unit (0.1 micron microfiltration membrane) and carries out the micro-filtration degerming and remove fat.Filtrate is transported to ultra-filtration membrane unit (the 30K molecule is by amount) for 136 liters and carries out desalination and concentration.Receive 0.52 liter of concentrated solution.Adopt freeze-drying method to get light brown green dry powder 351 grams, moisture 4.3%, total protein is 9 3.2%, ash content 2.6%.
Embodiment two: 996 liters of skimmed milks, and pH6.7, temperature 6.6 degree C total proteins are every liter of 32.9 gram, and BLP concentration is 378 milligrams every liter, and BLF concentration is 129 milligrams every liter.Adopt the operational path of Fig. 1, and produce monomeric BLP albumen and monomeric BLF albumen by the step described in the explanation.SPEC70 resin stirred pot absorption 115 minutes is housed in the adding, and stirring velocity is 60rpm.After collecting the milk of absorption, with RO water wash resin.Adding first step salt solution is separated for (2.4%) 90 liter and is told 36 minutes, stirring velocity 30rpm.Collection first step BLP separates and tells liquid and add 2.3% salt water wash resin 3 times.The merging leacheate is separated to BLP and is told back microfiltration membrane degerming and fat in the liquid.
At the BLP micro-filtration simultaneously, adding 86 liter of 4.9% salt solution carries out BLF and separates and tell stirring velocity 36rpm in stirred pot.Separate the 60 minutes time of telling.Collection is separated and is told to use RO water wash resin behind the liquid.Merge leacheate to separate tell in the liquid altogether 151 liters.Being transported to the micro-filtration unit carries out degerming and removes fat.Filtrate is transported to ultra filtration unit (30K molecule by amount) and adds water carries out desalination (wash ionic strength and be lower than 1.6 milli west doors) and is concentrated into 1% protein concentration molecule by amount) and add water and carry out desalination (wash ionic strength and be lower than 1.6 milli west doors) and when being concentrated into 1% protein concentration (7.6 liters), being transported to the CM resin container and removing foreigh protein removing.Deliver to after continuing then to be concentrated to 0.39 liter with ultra-filtration membrane the lyophilize unit dry rose pink powder 80.5 grams.Powder is moisture 3.1%, and ash 1.1%, total protein are 96.5%.
BLP behind the micro-filtration continues to concentrate after carrying ultra filtration unit to add water desalination (wash ionic strength and be lower than 1.6 milli west doors).Collect 0.73 liter of BLP concentrated solution.With freeze-drying dry brown-green powder 351 gram, powder is moisture 4.2%, ash 2.1%, total protein are 94.1%.
Claims (10)
1, a kind of method of from milk or whey water, extracting high purity protein, the stirred pot that its adopt to place Zeo-karb adsorbs, desorbs, lactoferrin, newborn Bovinelactoperoxidase and both mixtures thereof in separation and Extraction milk or the whey liquid, and is aided with membrane separation technique and it is separated concentrates and highly purified.
2, method according to claim 1 is characterized in that the adjustable stirring arm rotating speed of stirred pot is 20-100rpm; The jar inner side-wall has the eddy current of increasing baffle plate, and jar end is provided with the resin bed of band screen plate, and 8-12cm highly locates to be provided with quick water conservancy diversion suction filtration pipe above the resin layer on the resin bed.
3, method according to claim 1 is characterized in that being provided with in the jar the various transmitter pH meters of controlling for automatically, the weight meter, and electric conductivity is taken into account thermometer, and the pot liquid holding temperature is 3-160 ℃.
4, method according to claim 1 is characterized in that resin is the SPC70 resin, and number of times that resin absorption desorbs and condition can be set according to the requirement of variant production and product purity; The adsorption time of resin is 20-180 minute, and adsorption temp is at 3-30 ℃.
5, method according to claim 1, the adsorption time that it is characterized in that resin is 35-60 minute.
6, method according to claim 1 is characterized in that the employing two-stage salt concn that desorbs of resin desorbs lactoferrin, newborn Bovinelactoperoxidase.First step salt concn is 1-2.5%, desorbs newborn Bovinelactoperoxidase, and the time of desorbing is 30-100 minute; Second stage salt concn is 3.5-7%, desorbs lactoferrin, and the time of desorbing is 30-100 minute; The salt concn that desorbs newborn Bovinelactoperoxidase and lactoferrin mixture is to be 30-100 minute the 3-7% time of desorbing; The stir speed (S.S.) that desorbs is 20-100rpm.
7, method according to claim 4 is characterized in that it is sodium salt, calcium salt that resin desorbs used salt, sylvite; Resin need not regeneration activating after absorption and desorbing technological process, can be for the usefulness of next batch absorption as long as be lower than 1.6ms with reverse osmosis water rinsing resin to its ionic strength after desorbing the technology end.
8, method according to claim 1 is characterized in that microfiltration membrane that membrane separation technique uses the 0.05-0.1 micron with lactoferrin, newborn Bovinelactoperoxidase separate the attached liquid degerming, remove remaining fat and other small-particle; Ultra-filtration membrane with 1K-30K molecular interception amount carries out desalination and concentrated.
9, method according to claim 4 is characterized in that highly purified lactoferrin is that the lactoferrin dope that will desorb desalination and concentration through the absorption of the described method of claim 8 is separated the concentrated acquisition that echos ultra-filtration membrane through the absorption of a weak-type sun resin again.
10, method according to claim 1, it is characterized in that with adsorb, desorb, the lactoferrin dope of desalination and concentration, breast Bovinelactoperoxidase dope and lactoferrin thereof, newborn Bovinelactoperoxidase mixing dope carries out vacuum dehydrating at lower temperature or low temperature spray drying becomes powder mixture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100952443A CN100358916C (en) | 2006-03-06 | 2006-03-06 | Method for extracting high purity protein from cow milk or soybean waste water |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100952443A CN100358916C (en) | 2006-03-06 | 2006-03-06 | Method for extracting high purity protein from cow milk or soybean waste water |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1800200A CN1800200A (en) | 2006-07-12 |
| CN100358916C true CN100358916C (en) | 2008-01-02 |
Family
ID=36810444
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100952443A Active CN100358916C (en) | 2006-03-06 | 2006-03-06 | Method for extracting high purity protein from cow milk or soybean waste water |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100358916C (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101362792B (en) * | 2008-09-25 | 2011-07-20 | 上海交通大学 | Affinity separation polymer of lactoferrin and affinity purification method of lactoferrin |
| CN102898516A (en) * | 2012-10-26 | 2013-01-30 | 浙江大学 | Method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology |
| CN106008703B (en) * | 2016-07-06 | 2019-11-22 | 方雅悯 | A method of extracting iron saturation degree lactoferrin low with preparation high-purity from cow's milk |
| CN106377762A (en) * | 2016-08-24 | 2017-02-08 | 方雅悯 | Application of bovine lactoferrin and zymolytes thereof in products for protecting stomach and liver |
| CN109011821A (en) * | 2018-09-18 | 2018-12-18 | 安徽源森生物科技有限公司 | It is a kind of for manufacturing the filter of rice protein |
| CN112979785B (en) * | 2021-04-25 | 2021-12-28 | 黑龙江默赛东科技有限公司 | Method for preparing high-purity lactoferrin |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0717875A (en) * | 1993-02-09 | 1995-01-20 | Kyodo Nyugyo Kk | Production of iron-lactoferrin powder |
| CN1486989A (en) * | 2003-08-21 | 2004-04-07 | 安徽农业大学 | Technological process of separating and purifying lactoferritin from cow colostrum |
| EP1466923A1 (en) * | 2003-04-01 | 2004-10-13 | Snow Brand Milk Products Co., Ltd. | Method for producing lactoferrin |
| CN1557494A (en) * | 2004-02-04 | 2004-12-29 | 高春平 | Preparation of microorganism resisting biological active component and its application |
| CN1557950A (en) * | 2004-02-04 | 2004-12-29 | 高春平 | Method of preparing lactoferritin and lactoperoxidase |
-
2006
- 2006-03-06 CN CNB2005100952443A patent/CN100358916C/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0717875A (en) * | 1993-02-09 | 1995-01-20 | Kyodo Nyugyo Kk | Production of iron-lactoferrin powder |
| EP1466923A1 (en) * | 2003-04-01 | 2004-10-13 | Snow Brand Milk Products Co., Ltd. | Method for producing lactoferrin |
| CN1486989A (en) * | 2003-08-21 | 2004-04-07 | 安徽农业大学 | Technological process of separating and purifying lactoferritin from cow colostrum |
| CN1557494A (en) * | 2004-02-04 | 2004-12-29 | 高春平 | Preparation of microorganism resisting biological active component and its application |
| CN1557950A (en) * | 2004-02-04 | 2004-12-29 | 高春平 | Method of preparing lactoferritin and lactoperoxidase |
Non-Patent Citations (2)
| Title |
|---|
| 乳过氧化物酶的分离纯化和酶学性质研究. 芦蓉蓉等.食品科学，第27卷第2期. 2006 * |
| 牛初乳中乳铁蛋白的分离及纯化. 凌雪萍等.工艺技术，第23卷第12期. 2002 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1800200A (en) | 2006-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100358916C (en) | Method for extracting high purity protein from cow milk or soybean waste water | |
| CN101619308B (en) | Preparation method for extracting lysozyme from egg white | |
| CN101603038B (en) | Preparation method of lysozyme | |
| JPH03502921A (en) | Method for extracting pure fractions of lactoperoxidase and lactoferrin from whey | |
| CN101491287A (en) | Method for extracting lactose and lactoalbumin from whey and producing formulation milk powder | |
| KR20190135489A (en) | Cell Culture Purification | |
| CN104672328A (en) | Production method of human antithrombin III | |
| CHEN et al. | Microfiltration affinity purification of lactoferrin and immunoglobulin G from cheese whey | |
| CN104450639B (en) | A kind of method using ultrafiltration auxiliary aqueous two phase extraction technique extraction lactoperoxidase | |
| CN100558242C (en) | A kind of from colostrum the method for inductrialized separation of purified lactoferrins | |
| CN103013951B (en) | Method for extracting and purifying wheat germ lipase | |
| CN102898516A (en) | Method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology | |
| CN100480378C (en) | Process for producing lactoperoxidase | |
| CN102676475A (en) | Method for extracting muramidase from egg white | |
| CN103275943B (en) | Method for extracting superoxide dismutase from pig spleen | |
| CN103044569A (en) | Wolfberry extract containing wolfberry acid and preparation method thereof | |
| CN104327175B (en) | A kind of method for separating antibacterial peptide | |
| CN104109204A (en) | Method for separating and purifying recombinant human lactoferrins from paddy rice seeds | |
| CN107446905B (en) | Method for purifying recombinant human lysozyme | |
| CN102268418B (en) | Method for purifying lysozyme from milk | |
| CN1663961A (en) | Technology for separating and purifying lactoferritin from cow colostrum | |
| CN109134622A (en) | The multistep continuous Integration purification process of foot-and-mouth disease virus antigen | |
| CN1321126C (en) | Process for preparing high purity acarbose | |
| CN109705208A (en) | A kind of technique of single step chromatography preparation high-purity vWF ELISA | |
| CN104818310A (en) | Method for rapidly enriching active peptides in aquatic protein hydrolysate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee | ||
| CP02 | Change in the address of a patent holder |
Address after: 310007, room 8, No. 402, Hexi, Xihu District pine field, Hangzhou, Zhejiang Patentee after: Fang Yamin Address before: 225300 science and technology center, Gulou South Road, Jiangsu, Taizhou 348, China Patentee before: Fang Yamin |
|
| ASS | Succession or assignment of patent right |
Owner name: HANGZHOU LINGFAN BIOTECHNOLOGY CO., LTD. Free format text: FORMER OWNER: FANG YAMIN Effective date: 20140116 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 310007 HANGZHOU, ZHEJIANG PROVINCE TO: 311100 HANGZHOU, ZHEJIANG PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20140116 Address after: Green Ting Road Yuhang District Cang Qian street of Hangzhou city Zhejiang province 311100 No. 1 Building 1 Room 215 Patentee after: Hangzhou sailing Biotechnology Co., Ltd. Address before: 310007, room 8, No. 402, Hexi, Xihu District pine field, Hangzhou, Zhejiang Patentee before: Fang Yamin |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160708 Address after: 257347 Lu Yidong, king of Dongying agricultural hi tech industry demonstration area, Shandong Patentee after: Dongying Ruida Biological Technology Co., Ltd. Address before: Green Ting Road Yuhang District Cang Qian street of Hangzhou city Zhejiang province 311100 No. 1 Building 1 Room 215 Patentee before: Hangzhou sailing Biotechnology Co., Ltd. |