CN100417329C - Complete separating process for fresh liquid milk - Google Patents
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- CN100417329C CN100417329C CNB2005101289170A CN200510128917A CN100417329C CN 100417329 C CN100417329 C CN 100417329C CN B2005101289170 A CNB2005101289170 A CN B2005101289170A CN 200510128917 A CN200510128917 A CN 200510128917A CN 100417329 C CN100417329 C CN 100417329C
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Abstract
The present invention relates to an integrated separation process for fresh liquid milk. Centrifugation, ultra-filtration, ion exchange, ion exclusion separation and other bioseparation techniques can be used for continuously and completely separating fresh liquid milk so as to obtain cream, cheese, lactoferrin, macromolecular glycopeptide, the mixture of beta-lactoglobulin and alpha-lactalbumin, immunoglobulin G, gelatin whey protein, lactulose syrup, crystalline lactose and milk salt powder, which are used as important added components to be widely used in the production fields of milk foods, milk health products, medicines, fodder, etc. The separation process can be applied to cow milk, yak milk, buffalo milk, goat milk, etc., and a method is provided for processing cow milk and sheep milk in remote regions. Crystallized mother liquor is treated through the ion exclusion separation finally in the separation process; consequently, not only the milk salt powder can be produced, but also the discharge of a large amount of high BOD waste water is basically eliminated; environmental pollution sources are eliminated, and a contribution is made to environmental protection.
Description
Technical field
The present invention relates to the process technology of liquid milk, especially about complete separating process for fresh liquid milk.
Background technology
Popular dairy products-full-cream liquid milk, liquid yogurt and milk powder are Chinese in recent years dairy industry kinds with fastest developing speed.Through after the rapid growth of several years, speedup obviously slows down.In fact, the main outlet of milk consumption figure is the deep processing that is used for dairy products, for example cream, cheese, lactose, whey product, milk deli and other industrial use.Domestic seldom dairy products enterprise can produce solid and semisolid dairy products deep processed product, i.e. cream, cheese and whey product.
The challenge that 21 century food industry is faced is how to satisfy the most ripe, the most rich and the most fastidious consumer of a group on the human history.U.S.'s " food technology " magazine has been concluded the 21 centurys ten big propensity to consume (FoodTechnology, Vol.55, No.4,38-58,2001), and promptly 1) " convenient culinary art " food; 2) super delicious food and exquisiteness; 3) nutrient balance food; 4) " flatly " food; 5) novel family meals; 6) food heavily liked of child; 7) fresh and light; 8) vary the diet; 9) health food; 10) totally, pure natural and safety.High-quality and novel food products additive, food component and health food component are one of the most important industry that can satisfy 21 century consumer new demand.And fresh milk is these additives and adds one of most important material of component.In in the past 5 years, the international market is to the speed increment with 5-10% in demand every year of milk food and health products component.
Lactulose claims that again milk ketose (Lactulose) is a kind of cellobiose, is transformed through chemical isomerization by lactose.Research has in recent years found that lactulose has many physiology and pharmacological function, for example the promotion factor of Bifidobacterium, reduce intestinal gas generation, increase physical efficiency, increase sclerotin intensity, alleviate and treat chronic constipation, alleviate hyperammonemia and liver cerebral lesion, anti-endotoxin effect and prevention operation back jaundice complication, prevention kidney failure, suppress for the second time cholic acid, kill salmonella, hypoglycemic, reduce urinary system and respiratory system communicate illness, prevent (Mizota such as colonic adenoma recurrence, T. (1996) Bulletin of the IDF, 313,43-48).
Milk contains the casein about 2.6%, accounts for about 80% of gross protein.Casein is the Main Ingredients and Appearance of the main food-cheese in west/cheese (Cheese).Except cheese, casein can also be made casein (casoid flour), casein sodium and calcium caseinate.The casein product is as food additives and other industrial uses.With the casein is raw material, can also make nutritional preparations such as caseinhydrolysate and calcium polypeptide.
Whey protein (Whey Protein) is meant the slurries protein after (ox) milk is removed casein.Whey protein is not single albumen, but by the protein mixture of many different molecular weights and function.Following table 1 is listed main whey protein component (Kinsella, JE ﹠amp; Whitehead, DM, Adv.Food Nutr.Res.33:343-438,1989).Lactotransferrin is in the Chinese invention patent (patent No.: detailed description is arranged ZL02107373.2).
Table 1
| The whey protein component | Account for the ratio of whey total protein, % | Molecular weight (dalton) |
| Lactoglobulin | 54 | 14,200 |
| Lactoalbumin | 23 | 18,600 |
| Bovine serum albumin(BSA) | 6 | 66,000 |
| Immunoglobulin (Ig) | 6 | 150,000-1,000,000 |
| |
1 | 7,000 |
| |
2 | 76,000 |
| |
2 | 32,000-1,000,000 |
| Growth factor polypeptide and other polypeptide | 6 | ＞100,000 |
The value performance of whey protein is aspect three.The firstth, nutritive value, the essential amino acids content of whey protein is the highest in all edible proteins.It is 104 that its protein biology is worth (BV), is higher than 100 of egg albumen, and 80 of beef protein is caseic 77,74 of soybean protein, 54 of 59 and wheat gluten of rice gluten.The secondth, the physicochemical properties of its high-quality can be used as the food excipient, for example foaming, emulsification, gel etc.The 3rd, also be most important be exactly that whey protein contains many compositions with health care, for example immunoglobulin (Ig), lactotransferrin, organized enzyme and Porcine HGF.Many researchs reported whey protein can reduce the incidence of disease of colon cancer, can increase the human body glutathione content, increase muscle anabolism, reduce body weight.
Topmost protein composition is lactoglobulin (Lactoglobulin) in the whey protein, secondly is lactoalbumin (Lactoalbumin).The lactoglobulin body contains many binding sites as a kind of transport protein can carry inorganic salts, liposoluble vitamin and fat (Hambling, SC et al, In Advanced DairyChemistry Vol.1.Proteins. (Fox, P.F., ed.), pp.141-190.ElsevierApplied Science.London and New York, 1992).The amino acid evaluation of estimate of lactoalbumin is high, is rich in lysine, leucine, threonine, tryptophan and cystine.The bovine serum albumin(BSA) (Bovine serum albumin) that contains flow control three also has the binding ability of good essential amino acid evaluation of estimate and fat and aliphatic acid.The sex change bovine serum albumin(BSA) can reduce the incidence of disease (Strand, FT, US patent 5473050,1995) of insulin type diabetes and autoimmune disease.So the component of lactoglobulin, lactoalbumin and bovine serum albumin(BSA) has the application different with whey protein, can more effectively be applied to the baby milk goods such as this component.
Immunoglobulin (Ig) accounts for about 6% in the whey gross protein.The immunoglobulin (Ig) in Ruzhong has three kinds of hypotypes, immunoglobulin G (G
1And G
2), immunoglobulin A and immunoglobulin M, and in the cow's milk based on immunoglobulin G.Exist the purpose of immunoglobulin (Ig) to be to start the passive immunity system in the breast milk, the protection neonate avoids the infection of pathogenic bacteria.Milk immune globulin G has proved can be resisted effectively by Escherichia coli (Tacket, COet al, N.Engl.J.Med.318,1240-43,1988) and rotavirus (Davidson, GD etal, The Lancet, Sept 23,709-712,1989) the acute infection enterogastritis that causes.In addition, immunoglobulin (Ig) also has other physiologic function.
Big molecule glycopeptide (Glycomacropeptide) is the glycosyl part on the casein peptide chain.Renin is caseic at k-
105Phenylalanine
106The single-minded hydrolysis of methionine peptide bond produces big molecule glycopeptide and remains in the whey.Found that big molecule glycopeptide has many physiology and pharmacological function, comprised and reduce stomachial secretion, inhibition dental plaque and dental caries tooth, the promotion factor of bifidobacterium growth, control PKU, suppress blood platelet and solidify (El Salam, MH, etal, Int.Dairy is (4) J.6: 327-341,1996).Big molecule glycopeptide can also constrain appetite and change pigment production in the melanocyte by the release that stimulates pancreatic hormone cholecystokinin (CCK).The potential commercialization purposes of big molecule glycopeptide is extremely extensive.
The disaccharide that lactose is made up of a part galactolipin and a part glucose 1-4 glycosidic bond.Lactose is the main carbohydrate in nearly all mammiferous breast milk.Lactose has good physical chemistry, for example low sugariness, good flowability, agglomeration resistance, can adsorpting pigment and volatile flavor compounds, degree of wetting, decentralization, solubility, good compressing tablet.These character make lactose become irreplaceable additive of food industry, pharmaceutical industries and chemical industry and filler.Lactose can also promote the growth of chicken.Fat (Colbert, L.Decker, EA, J.Food Sci.56,1248-50,1991) can be put on weight and reduce to the lactose that adds 0.1-3.0% in the chicken feed.
Milk contains the necessary inorganic salts of life.The inorganic salts composition of milk comprises citrate, potassium, calcium, chlorine, phosphorus, sodium, sulphur, magnesium, and some trace elements.Except quite a few calcium and P elements are stayed in the cheese, most of milk inorganic salts are transferred in the mother liquor behind the lactose crystn.Crystalline mother solution is a kind of environomental pollution source, promptly can solve environmental problem from crystalline mother solution production milk salt, can also provide biological organic salt product for market.The inorganic salts of needed by human body are divided into two groups, i.e. volume inorganic salts for example copper, zinc, vanadium, cobalt, molybdenum etc. of sodium, potassium, calcium, phosphorus etc. and micro-inorganic salts for example.Milk salt almost contains required volume of all life and micro-inorganic salts.Moreover, the content of these inorganic salts has balance preferably, sodium-potassium balance for example, calcium phosphorus balance.So milk salt can also be inorganic salts and additive agent electrolyte as the salt that saline taste is arranged not only.
Summary of the invention
For this reason, the invention provides a kind of complete separating process for fresh liquid milk.Purpose is the large-scale commercial applications production technology that provides a kind of integral body to utilize liquid milk for dairy industry.This integrated technology not only can produce continuously a series of in milk food, high added value breast system health products and medicinal precursor in the interpolation component of demand, very environmental protection does not simultaneously have a large amount of high BOD discharge of wastewater.The present invention also is specially adapted to the milk source of some non-general marketplaces, for example yak milk, buffalo milk and Goat Milk.These milk sources all produce in the backward areas of China and are not accepted by most consumers.If but breast system component is made in deep processing, its nutrition, health care and functional value never are lower than milk.
The fresh liquid milk that uses in the separating technology of the present invention can be milk, yak milk, buffalo milk or Goat Milk.Complete separating process for fresh liquid milk of the present invention can separate processing and make 10 kinds of components such as cream, cheese/casein, lactotransferrin, big molecule glycopeptide, lactoglobulin and lactoalbumin mixture, immunoglobulin G, gel lactalbumin, Cephulac, crystallization lactose and milk salt powder.
Complete separating process for fresh liquid milk flow chart of the present invention is referring to Fig. 1.Its technology comprises the following steps:
(1) separation makes cream, cheese and lactotransferrin
1a) because the height and the seasonal variations of dairy bread difference, breeding technology, the component content of full-cream fresh milk fluctuates in certain scope: total solid 10.5-14.5%, butterfat 2.5-6.0%, protein 2.9-5.0%, lactose 3.6-5.5% and 0.6-0.9% inorganic salts.Prepare cream and cheese is mature technology from full-cream fresh milk.
1b) basic principle of cream preparation is by centrifugal separator butterfat and other materials to be separated.Basic technological operation obtains rare cream and defatted milk with cream separator after comprising full-cream fresh milk pasteurization.Rare cream again through beat, with salt and moulding, promptly make fatty 80%, the standard butter cream of salt 2%.
1c) basic principle of cheese preparation is by liquid milk acidifying (pH4.5) or the cohesion of interpolation renin are precipitated casein then.The cheese product has many different kinds.What the present invention described is to add the caseic method of renin coagulative precipitation in defatted milk, because have only with the whey liquid after the renin processing just to contain big molecular saccharides polypeptide.Defatted milk is warming up to 35 ℃, adds 0.005-0.02%CaCl then
2, 0.03% nitre, 1: 10,000-1: the liquid rennet of 15,000 intensity, stir and stop after 2 minutes, it is condensed.Cut the curdled milk face with cutter after 30 minutes, stir and be heated to 60 ℃ of enzymes that go out.Open the tap of cheese vat then, emit whey liquid, wet cheese dehydration, be dried to moisture and be lower than 10%, obtain casein after milling; Or the wet cheese of dehydration dissolves with 2.5 moles NaOH solution, and the pH of solution is about 6.7, and drying obtains water miscible casein-sodium pulvis; Maybe will wet after the cheese moulding, make cheese (Cheese) food through fermentation or other processing.
1d) whey liquid of preparation renin cheese generation is used to prepare lactotransferrin, and Chinese patent (patent No.: ZL02107373.2) describe by existing detail.This technology is separated lactotransferrin and whey liquid by step precipitation method.This processing step produces residual whey liquid for the first time.
(2) separation makes big molecule glycopeptide (Glycomacropeptide)
2a) residual whey liquid approximately contains protein 0.7% for the first time, lactose 4.8% and inorganic salts 0.8%, and big molecule glycopeptide 0.007% (70 mg/litre).
2b) big molecule glycopeptide or other protein, as amphoteric compound, can be positively charged under different pH values, negative electrical charge and not charged (isoelectric point, pI).When pH when isoelectric point is above, the anion exchange resin of positively charged can adsorb electronegative protein or polypeptide.When pH when isoelectric point is following, electronegative cationic ion-exchange resin can adsorb positively charged protein or polypeptide.Big molecule glycopeptide is a kind of very acid (the pI value is very low) and hydrophilic polypeptide, when pH4.7 is following, the main equal positively charged of whey protein component or in isoelectric point, and in this pH scope, big molecule glycopeptide is electronegative, thereby can suppose to be adsorbed by anion exchange resin single-mindedly.
2c) to be used to adsorb the ion exchange resin of big molecule glycopeptide be Rohm in the present invention; The anion macroporous ion exchange resin of Haas company, Amberlite IRA93 type for example, the matrix of resin is polystyrene-divinylbenzene, the ion exchanging function group is " N-(R)
2".Amberlite IRA93 type resin with 0.5 mole of HCl and the regeneration of 0.5 moles of NaOH, spends deionised water to pH neutrality before use again, with the acetate buffer solution balance of 0.05 mole of pH3.5-4.7, and dehydration then.The pH value of residual whey liquid is for the first time transferred to 3.5-4.7 with the HCl of 6 equivalents.Use gap stirring-type ion exchange resin absorption method, whey liquid and resin in stirred reactor suction-operated 30-60 minute, the for the first time residual whey liquid of every 100ml uses 10-50 gram resin, and the filtrate of filtration is next step and separates the residual whey liquid second time that uses.After polymeric adsorbent spends deionised water, big molecule glycopeptide with the absorption of 0.5-1.0 mole NaCl wash-out, eluent is with 5, the polyether sulfone of 000 dalton's molecular cut off (Polyethersulfone) milipore filter is ultrafiltration or use electrodialysis desalination on the cross-current ultrafiltration apparatus, vacuum concentrates then, obtains big molecule glycopeptide powder after the freeze drying.The separation yield is 65-75%, and purity is 50-60%.This processing step produces residual whey liquid for the second time.
2d) identify that with SDS-PAGE the composition of Amberlite IRA93 absorption is exactly big molecule glycopeptide.Electrophoresis pattern (Fig. 2) shows that IRA93 anion exchange resin eluent has clear and several fuzzy protein spectras.The theoretical molecular of big molecule glycopeptide is 7000 dalton.The performance molecule of other research (Kawasaki, Y, et al.Milchwissenschaft, 48,191-6,1993) demonstration glycopeptide is measured certainly pH value.At pH7, the molecular weight of glycopeptide is 2-5 ten thousand dalton, and at pH3.5, then is 1-3 ten thousand dalton.Chief reason might be because the self-polymerizing power of big molecule glycopeptide is very strong.
2e) the big molecule glycopeptide of the most direct proof Amberlite IRA93 selective absorption evidence be the amino acid content of protein and the result of sequence analysis.Known 8-10% trichloroacetic acid can precipitate all whey albumen but can dissolve big molecule glycopeptide (Milchwissenschaft 47 (11), 688-693,1992 for Kawakami H, et al).The adsorption sample of Amberlite IRA93 resin is handled precipitation with 10% trichloroacetic acid remove foreigh protein removing, supernatant is removed the trichloroacetic acid postlyophilization with molecular weight cutoff value 3500 daltonian dialysis membranes and is used for amino acid content and sequence analysis.The resin adsorption sample records the terminal sequence that plays 38 residues of N-: Met Ala Ile Pro Pro Lys Lys Asn Gln Asp Lys Thr Gln Ile Pro Thr IleAsn Thr Ile Ala Ser Gly Gln Pro Thr Ser Thr Pro Thr Ile Gln Ala ValGln Ser Thr Val
Compare with the sugared polypeptide amino acid order 1-38 residue of existing document (Sleigh R.In PhD thesis, University of NSW, Australia, P47,1982) report, fit like a glove.
What above-mentioned evaluation clearly illustrated that the absorption of IRA93 anion exchange resin is big molecule glycopeptide component.
(3) separation makes beta lactoglobulin and ALA mixture
3a) being used to separate the residual whey liquid continuation separation second time that obtains behind the big molecule glycopeptide has two kinds to select technologies, and one of them is that obtain whey protein matter component contains beta lactoglobulin and ALA mixture and immunoglobulin G component.The isoelectric point of ALA, beta lactoglobulin and bovine serum albumin (pI) be respectively 4.8,5.3 and 5.1, and the isoelectric point of immunoglobulin G family is in the scope of 7.7-8.3.When for the second time the pH of residual whey liquid and ion exchange resin transferred to 7.5, the anion exchange resin of positively charged can adsorb electronegative ALA, beta lactoglobulin and bovine serum albumin, and lets slip positively charged immune globulin Bai nationality.
3b) the present invention adopts for example Rohm ﹠amp of anion macroporous ion exchange resin; The Amberlite IRA93 type resin of Haas company is used to separate beta lactoglobulin and ALA mixture and immunoglobulin (Ig).The Amberlite IRA93 type resin of having regenerated, with the phosphate buffer balance of 0.05 mole of pH7.5, dehydration then.The pH value of residual whey liquid is for the second time transferred to 7.5 with the NaOH of 6 equivalents.The consumption of anion exchange resin (gram resin/100 milliliter whey liquid) is 20,50,100.Conclusion (of pressure testing) sees Table 2 and table 3, and table 2 shows that Amberlite IRA93 type weak anionic macroporous ion exchange resin whey liquid mainly optionally adsorbs ALA, beta lactoglobulin when pH7.5, but adsorbs immunoglobulin G less.Table 2 has also shown the influence of 4 processing (20 gram resin/100 milliliter whey liquid) to adsorption efficiency simultaneously, although resin has adsorbed in the whey liquid about 90% ALA and beta lactoglobulin when promptly handling for the third time, the loss of immunoglobulin G also reaches 40%.Table 3 item has shown the influence of different resins consumption to adsorption efficiency.
Table 2.IRA93 resin is repeatedly handled the influence to protein adsorption efficient
* the consumption of IRA93 resin: 20 gram resin/100 milliliter (residual whey liquid for the second time).
Table 3.IRA93 resin different amounts is to the influence of protein adsorption efficient
The result of table 2 and table 3 shows, uses higher resin demand, for example 100 restrains resin/100 milliliter whey liquids, and adsorption treatment efficient once for example 20 restrains resin/100 milliliter whey liquids than using lower resin demand for good, and absorption will be handled repeatedly.Per 100 milliliters of whey liquids used 100-200 gram resin when therefore the present invention operated.
3c) separating technology of the present invention be with the second time residual whey liquid pH be adjusted to 7.5 times, use AmberliteIRA93 anion exchange resin 100-200 gram resin/100 milliliter whey liquid, by gap stirring-type absorption method, be that whey liquid and resin after suction-operated 30-60 minute, are used for next step separating immune globulin G with the elimination of whey raffinate in stirred reactor.After polymeric adsorbent spends deionised water, with the protein of 0.5-1.0 mole NaCl wash-out absorption.Eluent is with the milipore filter ultrafiltration of 10,000 dalton's molecular cut offs or electrodialysis desalination and concentrate, and if desired, vacuum concentrates again, makes beta lactoglobulin and ALA mixture after spraying or the freeze drying.The yield of separating of ALA and beta lactoglobulin mixture is 60-70%, total protein content 93-98%, and wherein beta lactoglobulin and ALA account for the 65-69% and the 25-29% of gross protein respectively.
3d) SDS-PAGE is used in the evaluation of beta lactoglobulin and ALA, and concrete grammar is identified referring to the electrophoresis of big molecule glycopeptide.The protein spectra that dyes on the gel is optical density meter (Tianjin, island dual wavelength TCL scanner CS910) scanning carrying out quantitative analysis again.
(4) separation makes immunoglobulin G
4a) for the second time residual whey liquid passes through the filtrate after Amberlite IRA93 adsorbing separation goes out beta lactoglobulin and ALA mixture, also contains immunoglobulin G.This filtrate is transferred to pH4.5 with the hydrochloric acid of 6N, milipore filter ultrafiltration on the cross-current ultrafiltration apparatus with 100,000 dalton's molecular cut offs concentrates 10 times again, twice final vacuum of saturating filter concentrates, saturating filter is defined as that ultrafiltration concentrates retains and adds behind the water process of ultrafiltration again in the liquid, obtain the immunoglobulin G component after the freeze drying, see Table 4.
Table 4. separates the immunoglobulin G component that makes
| Milk is isolated the filtrate behind beta lactoglobulin and the ALA mixture | By the immunoglobulin G that makes the separation of milk filtrate | Colostrum is isolated the filtrate behind beta lactoglobulin and the α-lactoalbumin mixture | By the immunoglobulin G that makes the separation of colostrum filtrate | |
| Immunoglobulin G component purity (%) | 6.6 | 49.5 | ||
| Immunoglobulin G content (gram/100 gram dries) | 0.6 | 43.3 | 34.7 | 93.0 |
| The immunoglobulin G cycles of concentration | 72 | 2.7 |
Table 4 shows filtrate after milipore filter separates, and the purity of immunoglobulin G component is 43.3-93%, and cycles of concentration is 2.7-72 times.The milipore filter that this processing step processing back produces filters raffinate, is used for next step continuation separation and makes lactose or Cephulac.
4b) enzyme-linked immunosorbent assay (ELISA) is used in the evaluation of immunoglobulin G and quantitative.The principle of immunoassays is the strongest antigen-antibody reactions of selectivity.At this, antigen is the immunoglobulin G in the sample, and antibody is anti-cattle immune globulin G reagent.The present invention adopts sandwich-antibody experimental method, and antibody uses anti-cattle immune globulin G reagent of rabbit and the anti-cattle immune globulin G reagent of peroxidase connection rabbit of DAKOPATTS.See for details Coligen method of operating (Current Protocol in Immunology, New York:Green, 1991:2.1.2-2.1.16).So-called sandwich-antibody act, promptly earlier the anti-cattle immune globulin G reagent of rabbit is coated on the little hole wall of immunoassay plate, add sample then, immunoglobulin G combines with its antibody in the sample, other compositions then are cleaned, adding the anti-cattle immune globulin G reagent of peroxidase connection rabbit again combines with immunoglobulin G just as a sandwich, add i.e. 30% hydrogenperoxide steam generator of peroxidase substrate at last, durol methanol solution and pH6 acetate buffer solution carry out color development reaction (yellow), under 450 nanometers, use 3550 type micro plate analyzer (Microplate Reader) colorimetrics of Bio-Rad then, can automatically test out the content of immunoglobulin G in the sample again with related software.
(5) separate preparation gel whey protein
5a) the present invention handle residual whey liquid for the second time another to select technology be to prepare gel whey protein (see figure 1) from lactalbumin.This preparation technology's the first step is that the preparation protein content surpasses 90% PURE WHEY.For the second time residual whey liquid at first on the milipore filter device ultrafiltration be concentrated into and contain protein 15-20%.Concentrate is transferred to 3.4-3.6 with the HCl of 6 equivalents with the pH value, best pH3.5, the cationic ion-exchange resin Purolite C-150S type of pH3.4-3.6 of regenerating of packing into, per 100 milliliters of whey liquids use 140-200 gram resin, the matrix of resin is polystyrene-divinylbenzene, mechanical strength is big, and rigidity is big and arrange loosely between particle, is easy to the industrialization operation.At room temperature, stirring and adsorbing 30-60 minute, leach the processing raffinate.The protein that polymeric adsorbent adsorbs with 0.5-1.0 mole NaCl wash-out.Eluent is with the milipore filter device ultrafiltration desalination of 10,000 dalton's molecular cut offs and concentrate, and concentrates as the further vacuum of needs, obtains PURE WHEY after spraying or the freeze drying then, and its protein content is 90-95%.This processing step produces resin and takes off albumen processing raffinate, is used for the separation preparation of next step lactose or lactulose.
5b) in order to prepare gel lactalbumin liquid or pulvis, with PURE WHEY or directly be mixed with the 3-10% lactalbumin aqueous solution, with citric acid pH is transferred between the 3.5-4.5, preferably pH3.8 with the fluid whey protein milk, heating-up temperature 80-100 ℃, promptly make stabilizing gel lactalbumin slurries.Further can make the gel PURE WHEY with high pressure nozzle formula spray dryer.Lactalbumin through gelation can be slurries or pulvis, can be used as protein additive and is used for acidic juice beverage, acid jam and other acid food.Through the gel lactalbumin liquid of above-mentioned technology preparation, precipitation, layering, viscosity in 12 months, do not occur and reduce phenomenon thinning or that the viscosity increase is solidified.
(6) separation makes Cephulac
6a) the 4th step was isolated the raffinate after the milipore filter that produces behind the immunoglobulin G filters raffinate or the resin adsorption lactalbumin is handled in the 5th step) in mainly contain lactose, inorganic salts and a spot of nonprotein nitrogen, it is continued separation has two kinds to select technologies, one of them is that lactose is changed into lactulose, and then makes Cephulac.Montgomery and Hudson (Science, 69,556,1929) have found that at first lactose is to the structural rearrangement of lactulose under alkali condition.Subsequently, United States Patent (USP), US 3,707, and 534 (1972), US3,816,174 (1974) and US 4,536,221 (1985) have set up lactose basically to the isomerized condition of lactulose.The isomerized catalyst of lactose is an alkaline metal cpds, mainly comprises Ca (OH)
2, KOH, Na
2CO
3, NaOH, sodium bisuflide (NaHS), Mg (OH)
2Deng.Reaction temperature is about 40 ℃, and the corresponding 24 hours reaction time is not waited.Some are than the patent documentation in later stage, and for example US 5,034, and 064 (1991) and US6,214,124 (2001) have also described similar method.Yet these inventions all are to transform with the solution that pure lactose is prepared, and a large amount of ion exchange resin of adding come cessation reaction under reaction temperature.Resin is destroyed can not to be reclaimed, so the cost height.This method is also impracticable when heavy industrialization is operated.Coming modified milk fructose with impure solution is different with pure lactose solution in the grasp of process conditions.The situation that the present invention comes to this, the existence of impurity can hinder with milipore filter filter raffinate particularly resin take off albumen and handle the lactose concn of raffinate and be concentrated into more than 70%.Therefore to change into the process modification of lactulose as follows for the lactose in the technology of the present invention:
Whey raffinate after at first milipore filter being filtered raffinate or resin adsorption lactalbumin and handling is concentrated into and contains about lactose 30-40%, at 45-50 ℃ with cationic ion-exchange resin for example the sulfonic group strong-acid type PCR 553 or PCR 883 decationized Y sieves of Purolite company; Anion exchange resin is Rohm ﹠amp for example; The quaternary amine base strong base IRA401 of the company of Hass or 402 exhaustion of yin ion and decolourings; Then further use Rohm ﹠amp as needs; The quaternary amine base strong base macroporous anion exchange resin IRA900 or 938 of the company of Hass takes off residual protein and polypeptide; And then with solution concentration to containing lactose 75-80%, be 10-20% with the concentration adjustment of reforming catalyst alkali compounds in the concentrate, alkali compounds can be selected from NaOH, KOH, Ca (OH)
2, Na
2CO
3, NaHS or Mg (OH)
2Deng, about 80-100 ℃ of temperature heating, react after 30-60 minute, be quickly cooled to 50 ℃, add the sulfonic group strong-acid type cation exchange resin PCR553 or the PCR 883 of an amount of water and Purolite company or use the electrodialysis desalination cessation reaction with chilled brine.Can reduce galactolipin with the quick cooling of the chilled brine of 5-10% concentration in conversion reaction and occur, improve conversion ratio, the amount of resin of using after also can reducing reduces cost greatly.With solution concentration to 80 ° Bx, be cooled to 25 ℃ again, remove by filter the unreacted lactose of crystallization.Lactose reaches 50-60% to the conversion ratio of lactulose.Supernatant is condensed into the Cephulac of 80 Brix Scales (° Bx) again.This syrup contains lactulose more than 50%, and promptly lactulose accounts for the 62-80% of syrup total solid, and remaining composition is galactolipin and lactose.
6b) the mensuration of lactulose, use the standard gas chromatograph method of Sweeley to measure (Sweeley, etal.JACS, 85,2497-2507,1963), use colorimetric determination (Nagendra and Venkat, Food Chemistry, 43 after perhaps using the thin-layered chromatography separating lactulose, 399-402,1992).
(7) lactose crystn
Another selection technology that continues the raffinate after processing milipore filter filtration raffinate or resin adsorption lactalbumin are handled is to prepare lactose by crystallization, and this is very ripe technology basically.The raffinate that milipore filter is filtered after raffinate or resin adsorption lactalbumin are handled is concentrated into total solid 30% earlier, can add activated carbon, heating, filter, decolouring is concentrated into total solid about 64% with multi-effect vacuum evaporator again, makes lactose concn reach hypersaturated state.With concentrating the crystallizing tank that filtered solution is packed the strap clamp cover into and stirred, be warming up to 70 ℃, add the lactose crystal seed.Cooling while stirring folds lactose crystn then, with centrifuge or flame filter press crystallization lactose cream is separated with mother liquor.After crystallization lactose cream washes with water, last boiled bed drying, abrasive dust, promptly obtain the lactose powder, lactose content is about 90%.Be recrystallized after crystallization lactose cream adds the water heating for dissolving, can make the product of lactose content more than 98%.This processing step produces crystalline mother solution and is used for next step continuation separation.
(8) separation makes milk salt powder and lactose from crystalline mother solution
8a) crystalline mother solution is the very high environomental pollution source of BOD, contains unsegregated lactose and inorganic salts.The basis that mother liquor separates is to utilize ion exclusion to separate chromatographic column.The principle of ion exclusion separation: the functional group of ion exchange resin, particularly cationic ion-exchange resin (sulfo group SO for example
3 -) by the metal cation saturated after, promptly form RSO
3-Na
+A kind of like this ion exclusion system, but non-ionic compound can not be subjected to enter resin with repelling.This ion exclusion system can be used for non-ionic compound and ionic compound are separated.
8b) the present invention uses the sulfonic group strong-acid type cation exchange resin PCR 553 or the Rohm﹠amp of Purolite company; The quaternary amine strong base IRA401 anion exchange resin of Hass company.They are gel type polystyrene-divinylbenzene resin, 4% interlinkage degree and 0.3-0.4mm pearl molded dimension.
The different salt types of table 5. resin PCR 553 and IRA 401 are to the influence of Kd and α value
| Resin | The salt type | K d ** (Lac*) | K d ** (NaPi*) | K d ** (Ca*) | α *** (Lac/NaPi) | α *** (Lac/Ca) |
| PCR 553 PCR 553 PCR 553 IRA 401 IRA 401 | NH 4 Na K Cl CO 3 | 1.35 1.12 1.12 1.47 1.82 | 0.88 0.76 1 1 1.24 | 0.88 1 1.12 1.12 | 1.25 1.2 1.06 1.24 1.26 | 1.25 1.06 1 1.17 |
* Lac: lactose, NaPi:NaH
2PO
4, Ca:CaCl
2
* distribution coefficient (K
d) be defined as specific ion and nonionic solute ratio (K in the inside and outside concentration of ion exclusion resin
d=C
r/ C
1).Each solute all has specific K for different salt types
dValue.The K of two kinds of solutes
dDifference, represent that two kinds of solutes can separate on chromatographic resin.Difference is big more, and separation property is good more.K
dValue can also be used to indicating one group of eluted order of solute.K
dThe solute solute of value minimum is at first by wash-out.
K
d＝(V
n-V
o)/V
o
V
o=external void liquid volume between resin, i.e. void volume.
V
n=on the wash-out collection of illustrative plates, be eluted to peaked volume to solute from solute is just more eluted.
* * selects coefficient-α (Skoog and Leary, In Principles of Instrumental Analysis4th edn., Saunders College Publishing, pp.579-604,1992) be used to describe ionic compound-salt and polypeptide, with non-ionic compound-lactose, the speed difference when moving on chromatographic resin defines with following formula:
α＝K
d L/K
d I
K
d LBe the lactose distribution coefficient, K
d IIt is the distribution coefficient of ionic compound (salt and polypeptide).If α equals 1, lactose and ionic compound will appear in the eluent simultaneously, promptly can't separate.If α is greater than 1, ionic compound will be at first by wash-out.The α value is big more, and the separating effect between lactose and the ionic compound is good more.
The salt type of resin is a most important factor during ion exclusion separates.The present invention uses cationic ion-exchange resin PCR553NH
4Type, anion exchange resin IRA401Cl or CO
3Type.Table 5 has been showed NH
4-type and Cl and CO
3The comparison of the Na of-type and prior art and K-type separative efficiency (United States Patent (USP) 4,820,348,4,955, the 363 cationic ion-exchange resin ion exclusion methods that disclose a kind of PCR553Na and K-type are handled whey liquid in the pH5.5-7.9 scope).The result shows new Ying's salt type that the present invention adopts high 16-19% of separative efficiency than prior art salt type.
8c) method for preparing the ion exclusion resin is at first to use 0.5M HCl or NaOH and washing to convert resin (the open market product is Na type or K type) to free acid/alkali type, with the 0.5MNH of 4 times of resin volumes
4Cl, or 0.5M NaCl, or 0.5M Na
2CO
3With become the NH4 type after the resin treatment, Cl type or CO
3Type is washed to pH neutrality.Pack into carefully preparation in the post of resin there is not the chromatographic column of tomography.
8d) the present invention uses district's belt chromatography (Zonal Chromatography), after being about to crystalline mother solution raw material that pH transfers to 5.0-5.5 and pumping into chromatographic column, waits it all to imbed and pumps into eluent water again behind the resin bed and carry out wash-out.The flow velocity of mother liquor is regulated with pump, flow velocity be the 0.06-0.24 liter/hour.Feed volume is controlled at the resin bed volume of 2-14%.Temperature remains on 60-80 ℃ in case low temperature causes sugared crystallization to block chromatographic column during separation.The present invention can separate (referring to Fig. 4) well with lactose with inorganic salts.When chromatography, use the distribution of refractometer and conductivity gauge monitoring collection of illustrative plates, guarantee that lactose and inorganic salts component can thoroughly separate basically.The lactose component eluent of collecting is used for recirculation and gets back to the lactose crystn operation, and the inorganic salts component of collecting concentrates the back and makes milk salt powder with spraying or boiled bed drying.
8e) lactose is measured and is used refractometer, and lactose concn is in Brix Scale (° Bx).Activon 301 type conductivity gauges are used to measure inorganic salts.
Complete separating process for fresh liquid milk of the present invention separates that the cream, the milk that make are cruel, lactotransferrin, big molecule glycopeptide, lactoglobulin and 10 components such as lactoalbumin mixture, immunoglobulin G, gel lactalbumin, Cephulac, crystallization lactose and milk salt powder, in milk food, breast system health products purposes is very widely arranged.
Below table 6 list each component that the complete separating process that utilizes fresh liquid milk of the present invention gets in Application for Field such as milk food, breast system health products, medicinal thing, feed, beauty treatments.
Table 6
| Composition | Major function | Food | Baby food | Healthy food | Medicine/clinical | Animal doctor/feed | Beauty treatment/other |
| Cream | Smear fat | Be suitable for | Be suitable for | ||||
| Cheese/casein | Protein food and additive, industry adhesive | Be suitable for | Be suitable for | Be suitable for | Be suitable for | Be suitable for | |
| Big molecule glycopeptide | Physiology and pharmacological function | Be suitable for | Be suitable for | Be suitable for | Be suitable for | Be suitable for | |
| The mixture of lactoglobulin and lactoalbumin | Best amino acid evaluation of estimate; Good foaming and emulsification function | Be suitable for | Be suitable for | Be suitable for | Be suitable for | Be suitable for | Be suitable for |
| Immunoglobulin G | Opposing bacterium and virus | Be suitable for | Be suitable for | Be suitable for | Be suitable for | Be suitable for | |
| The gel lactalbumin | The homoamino acid evaluation of estimate; At pH 3.5-4.5 stabilising whey protein liquids | Be suitable for | Be suitable for | Be suitable for | Be suitable for | ||
| Cephulac | The promotion factor of fork bacillus | Be suitable for | Be suitable for | Be suitable for | Be suitable for | ||
| The crystallization lactose | Low sweetness sugars, good flowability and fillibility are used for food interpolation and tablet carrier, chicken growth stimulator | Be suitable for | Be suitable for | Be suitable for | Be suitable for | Be suitable for | |
| Milk salt powder | Salt and inorganic salts | Be suitable for | Be suitable for | Be suitable for |
Fresh liquid milk separating technology of the present invention has following advantage;
1) provide fresh liquid milk to separate and the complete production technology that combines of comprehensive utilization first, except milk, the present invention also is suitable for processing yak milk, buffalo milk and Goat Milk.For the backwoodsman raw material processing of China provides outlet.
2) the present invention is " integral type " integrated technique production, can continuously fresh liquid milk be separated the interpolation component of demand in numerous food product, health products or the medicine, be butter, casein, lactotransferrin, mixture, immunoglobulin G, gelation lactalbumin, Cephulac, lactose and the milk salt powder of big molecule glycopeptide, beta lactoglobulin and ALA.
3) between the best pH scope 3.5-4.5 of acidic beverages and food, the aqueous solution of lactalbumin is extremely unstable but pass through.But can process in the pH3.5-4.5 scope no layering and not have the new type gel lactalbumin component that solidifies by technology of the present invention, it can be used as the proteins additive of acidic beverages.
4. the present invention uses ion exclusion separation to handle the lactose crystn mother liquor, not only can produce milk salt powder, has eliminated the situation of a large amount of high BOD discharge of wastewater simultaneously basically, and environmental protection is contributed, and makes more manageability of factory.
Description of drawings
Fig. 1 complete separating process for fresh liquid milk flow chart of the present invention.
The SDS-electrophoresis pattern of the eluent of the big molecule glycopeptide of Fig. 2 IRA93 resin adsorption.
The SDS-electrophoresis pattern of Fig. 3 ALA and beta lactoglobulin mixture.
Road 1: residual whey liquid for the second time, road 2: immunoglobulin G, road 3: ALA and beta lactoglobulin mixture, road 4: the standard molecular weight of each component.
La: ALA, Lg: beta lactoglobulin, BSA: bovine serum albumin, 1gG-H: immunoglobulin (Ig)
G heavy chain, IgG-L: IgL.
Fig. 4 lactose separates the wash-out collection of illustrative plates with inorganic salts.
Chromatography condition: resin: PCR 553 NH4-types; Resin bed height=110cm; Resin volume=150ml; Flow velocity=1ml/min; Temperature=60 °, sample pH value 5.5; Curve:----NaH
2PO
4---lactose.FV: feed volume; RBV: resin bed volume.Ce: the concentration of material in eluent; Cf: the concentration of material in mother liquor, Ce/Cf represents the degree of material wash-out.
The specific embodiment
Embodiment 1: the separation preparation of butter
1000 liter whole milks contain 3.8% butterfat, 3.5% protein, 4.8% lactose and 0.9% inorganic salts.Milk is cooled to 4 ℃ through 72 ℃ of pasteurizations 20 seconds, obtains 900 liter skim milks (creaminess 0.05%) with the standard cream-milk separator, 92 kilograms of rare cream (creaminess 40.6%).Next process is standby behind the skim milk pasteurization.Rare cream through 95 ℃ of pasteurizations, 62 ℃ of vacuum outgas, add operations such as 439 gram salt, cream stirring, obtain 44.4 kilograms of butter, creaminess 85%, salt 1%, and the buttermilk about 45 kilograms (buttermilk).
Embodiment 2: the separation preparation of casein
900 liter pasteurised skimmed milk behind 1 fen butter of leaving away of embodiment are inserted cheese vat, temperature is remained on 35 ℃, add 180 gram CaCl then
2, 279 gram nitre, 270 milliliters of liquid renins (1: 10,000-1: 15,000 intensity), stir and stop after 2 minutes, it is condensed.Cut the curdled milk face with cutter after 30 minutes, stir and be heated to 60 ℃ of enzymes that go out.Open the tap of cheese vat then, emit the whey liquid of 700 liters.Wet cheese continues the whey liquid that dehydration obtains 80 liters, obtains the whey liquid of 800 liters at last, and is standby behind the pasteurization.The wet cheese of dehydration is dried to moisture and is lower than 10%, obtains 26.6 kilograms of casein after milling.
Embodiment 3: the separation preparation of big molecule glycopeptide
Then embodiment 2800 liter pasteurized whey liquid are through the Chinese invention patent (patent No.: after method ZL02107373.2) is handled and separated lactotransferrin, time residual whey liquid of winning is inserted agitator tank, with 6N hydrochloric acid pH is transferred to pH4.7 while stirring, put into Amberlite IRA93 type resin (the Rohm ﹠amp of 180 kilograms of pH4.7 that regenerated then; Haas company product), at room temperature, stirring and adsorbing 30 minutes.Elimination gets residual whey liquid for the second time.After polymeric adsorbent washs with 50 liter deionized waters, the protein of 1 mole of NaCl wash-out absorption of 200 liters.The eluent of 200 liters is with the poly (ether-sulfone) ultrafiltration membrane of 5,000 dalton's molecular cut offs ultrafiltration and saturating filter desalination on the cross-current ultrafiltration apparatus, and vacuum is concentrated into 2 liters then, obtains the big molecule glycopeptide powder of 66.4 grams after the freeze drying, purity 59%.
Identify Amberlite IRA93 adsorption sample with SDS-PAGE.Use commercialization standard electrophoresis apparatus, gel slab, buffer solution (Gradipore product and operation sequence).Fixed solution contains ethanol 50%, acetic acid 10% and water 40%.The dyeing aqueous solution contains Coomassie blue 0.5% and amido colours 10B0.5%.3% acetum is used for decolouring.Electrophoresis pattern (Fig. 2) shows that IRA93 anion exchange resin eluent has clear and several fuzzy protein spectras.The theoretical molecular of big molecule glycopeptide is 7000 dalton.The performance molecule of other research (Kawasaki, Y, et al.Milchwissenschaft, 48,191-6,1993) demonstration glycopeptide is measured certainly pH value.At pH7, the molecular weight of glycopeptide is 2-5 ten thousand dalton, and at pH3.5, then is 1-3 ten thousand dalton.Chief reason might be because the self-polymerizing power of big molecule glycopeptide is very strong.The adsorption sample of Amberlite IRA93 is handled precipitation with 10% trichloroacetic acid remove foreigh protein removing, supernatant is removed the trichloroacetic acid postlyophilization with molecular weight cutoff value 3500 daltonian dialysis membranes and is used for amino acid content and sequence analysis.Amino acid content with amino-acid analyzer AAA-Waters Picotag system testing sample.Be transferred on the poly-difluoride membranes with the single bands of a spectrum of immunoblotting (instrument of Bio-Rad and S.O.P.), put into the full-automatic protein sequence analyzer of Applied-Biosystems-470A type and measure 38 amino acid residues of this polypeptide from amino terminal with big molecule glycopeptide.Be used for amino acid content and sequence analysis after the dialyzed sample freeze drying.The amino acid content of table 7 show sample and the theoretical value of sugared polypeptide are very approaching.
The comparison of amino acid content and glycopeptide theoretical value in the IRA93 resin elution liquid after table 7. is handled with trichloroacetic acid
* theoretical value is calculated from the primary structure of glycopeptide.
Terminal 38 residues of N-are only carried out in the amino acid sequence analysis of sample, and the sequence that the eluent sample records is:
Met Ala Ile Pro Pro Lys Lys Asn Gln Asp Lys Thr Gln Ile Pro Thr IleAsn Thr Ile Ala Ser Gly Gln Pro Thr Ser Thr Pro Thr Ile Gln Ala ValGln Ser Thr Val, with existing document (Sleigh R.In PH.D.thesis, University ofNSW, Australia, p47,1982) report sugared polypeptide amino acid order 1-38 residue compare, fit like a glove.What above-mentioned evaluation clearly illustrated that the absorption of IRA93 anion exchange resin is big molecule glycopeptide component.
The quantitative employing adsorption sample of big molecule glycopeptide is handled through 10% trichloroacetic acid precipitation and 3500 daltonian dialysis membranes earlier, then with improvement Biuret Method (Sleigh, R.In Ph.D.thesis, University of NSW, Australia, p 47,1982) the mensuration total protein content.
Embodiment 4: beta lactoglobulin and ALA mixture separate preparation
Then embodiment 3 inserts agitator tank with the 720 liters residual whey liquid second time, NaOH with 6 equivalents transfers to 7.5 with the pH value while stirring, the Amberlite IRA93 type resin of pH7.5 of having regenerated of packing into 720 kilograms, at room temperature, stirring and adsorbing 30 minutes leaches the whey raffinate.The Amberlite IRA93 that adds 360 kilograms of pH7.5 that regenerated in the whey raffinate again, stirring and adsorbing is elimination whey raffinate after 30 minutes.1080 kilograms of polymeric adsorbents with the washing of 2000 liter deionized waters after, the protein that adsorbs with 1 mole of NaCl wash-out of 2000 liters again.Eluent is with 10, the polyether sulfone of 000 dalton's molecular cut off (Polyethersulfone) milipore filter is ultrafiltration and saturating filter desalination and concentration on the cross-current ultrafiltration apparatus, vacuum is concentrated into 20 liters then, obtains 4.2 kilograms of beta lactoglobulins and ALA mixture powder after spraying or the freeze drying.The protein content 96% of protein mixture powder, wherein beta lactoglobulin and ALA account for 68% and 28% of gross protein respectively.
Identify that with SDS-PAGE the composition of IRA93 resin adsorption is beta lactoglobulin and ALA mixture, method records and the results are shown in Figure 3 electrophoretograms with embodiment 2.
Embodiment 5: the separation preparation of immunoglobulin G powder
Embodiment 6: the separation preparation of the lactalbumin of gelation
Handling another selection of residual whey liquid for the second time is preparation gelation lactalbumin (see figure 1).This preparation technology's the first step is that the preparation protein content surpasses 90% lactalbumin slurry or pulvis.The 720 liters whey raffinate second time is concentrated into 24 liters with polyether sulfone (Polyethersulfone) milipore filter ultrafiltration on the cross-current ultrafiltration apparatus of 10,000 dalton's molecular cut offs, contains protein 19%.Concentrate is inserted agitator tank, and the HCl with 6 equivalents transfers to 3.4-3.6 with the pH value while stirring, preferably pH3.5, regenerated the cationic ion-exchange resin Purolite C-150S type resin of pH3.5 of 38 liters of packing into, at room temperature, stirring and adsorbing 60 minutes leaches raffinate.It also can be used for next step and continues to separate preparation Cephulac or lactose crystn.Polymeric adsorbent is with after the 70 liter deionized waters washings, again the protein that adsorbs with 1000 liter 0.5-1.0 mole NaCl wash-outs.Eluent is with the poly (ether-sulfone) ultrafiltration membrane of 10,000 dalton's molecular cut offs ultrafiltration and saturating filter desalination and concentration on the cross-current ultrafiltration apparatus, and vacuum is concentrated into 20 liter lactalbumins slurry then, obtains 2.8 kilograms of PURE WHEYs after spraying or the freeze drying.The protein content 95% of albumen powder.
Preparation technology's second step is with 84 kg of water 2.8 kilograms of PURE WHEYs dissolves, or directly 20 liter lactalbumins slurry is transferred to the concentration of 3-4%, adds 577 gram citric acids again, thoroughly dissolves, at this moment between the pH value 3.6-3.8.Agitating heating is 14 minutes in 91.5 ℃ jacketed reaction pot.Promptly make the acid lactalbumin slurries of gelation after the cooling.As further obtaining the acid PURE WHEY of about 2 kilograms of gelations with high pressure nozzle formula spray dryer drying.
Embodiment 7: Cephulac separation preparation
Follow the filtration raffinate that embodiment 5 or embodiment 6 produce, first selection is to produce Cephulac.Three filtered solutions of 1800 liters that use-case 5 produces and the washings of example 4 merge, and contain about 33.5 kilograms of lactose, 4.9 kilograms of inorganic salts, 1.6 kilograms of protein (polypeptide and nonprotein nitrogen).At first filtered solution and washings mixed liquor are concentrated into 1000 liters, wherein 50% is used for the lactose crystn operation.500 liters are concentrated filtered solution be concentrated into 50 kilograms, PCR553 cationic ion-exchange resin and 20 liter Rohm ﹠amp 45-50 ℃ of elder generation with 20 liter Purolite companies with multi-effect vacuum evaporator; The desalination of IRA401 anion exchange resin process and the decolouring of the company of Hass are concentrated into 20 kilograms again, wherein contain promptly about 16 kilograms of lactose 80%.2.4 kilogram NaOH mixes with concentrate with the low amounts of water dissolving, is heated to about 90-95 ℃, reacts 60 minutes, is cooled to 50 ℃, adds PCR 883 cationic ion-exchange resins of an amount of water and 2000 gram Purolite companies, the desalination cessation reaction.With solution concentration to 80 ° Bx, be cooled to 25 ℃ again, remove by filter the unreacted lactose of crystallization, obtain the Cephulac of 13.5 kilograms of 80 ° of Bx at last, contain 7.9 kilograms of lactuloses, 1.2 kilograms of galactolipins and 1.6 kilograms of lactose.
Embodiment 8: the fractional crystallization of lactose
Other 500 liters among the embodiment 6 are concentrated filtered solution, can add 2% activated carbon, filter decolouring after 15 minutes, be concentrated into 31.5 kilograms with multi-effect vacuum evaporator again, contain total solid about 64%, make lactose concn reach hypersaturated state 100 ℃ of heating.With concentrating the crystallizing tank that filtered solution is packed the strap clamp cover into and stirred, be warming up to 70 ℃, add 0.2% lactose crystal seed.Use 10 ℃ of water quench to 15 ℃, cooling velocity per hour to be controlled at 2 ℃ then while stirring and make it to separate out lactose crystn.After 27 hours, crystallization lactose cream is separated with mother liquor, mother liquid obtainedly be used for next step and continue separate with centrifuge or flame filter press.Crystallization lactose cream obtains 11 kilograms of lactose powder after washing with 15 ℃ of deionized waters of 25 liters behind last boiled bed drying, the abrasive dust, lactose content is about 90%.Be recrystallized after crystallization lactose cream adds the water heating for dissolving, can make the product of lactose content more than 98%.
Embodiment 9: the chromatography of mother liquor
Then embodiment 8 contains 6.4 kilograms of lactose with about 20.5 kilograms mother liquor, 2.3 kilograms of inorganic salts, 0.7 kilogram of protein.The pH of mother liquor is transferred to 5.5 with the HCl of 6N, and the ion exclusion of packing into then separates chromatography and leans on separation, and the chromatography condition is as shown in table 8 below.Water wash-out is then at first collected the eluent of 156 liters, and this component contains inorganic salts and lactose, it is concentrated back drying obtain 2.1 kilograms of milk salt powder.Collect the eluent (seeing Table 9) of 100 liters from the 157th liter, this component contains lactose, turns back to the lactose crystn operation after being concentrated into 50 ° of Bx, or is used for making the lactose syrup.
Table 8 chromatography condition
| Resin | Purolite strongly acidic cation-exchange PCR 553, NH 4Type; Or strong basic type anion-exchange resin IRA401 Cl of Rohm﹠Hass company or CO 3Type |
| The chromatographic column diameter | 0.26 rice |
| The resin height of bed | 6 meters |
| Flow velocity | 1ml/ |
| Temperature | |
| 60℃ | |
| pH | 5.5 |
| Feed volume | 6.4% resin bed volume |
Table 9 eluent component is collected
| Component | Effluent volume, liter |
| The inorganic salts component | The 0-156 liter amounts to 156 liters |
| The lactose component | The 157-256 liter amounts to 100 liters |
Claims (8)
1. a complete separating process for fresh liquid milk is characterized in that comprising the following steps:
(1) will be from the fresh liquid milk branch cream of leaving away, the residual whey liquid first time that produces behind cheese and the lactotransferrin separates, residual whey liquid pH is adjusted to 3.5-4.7 for the first time, adopt anion exchange resin IRA93 type to adsorb big molecule glycopeptide, per 100 milliliters of whey liquids use 10-50 gram resin, stirring and adsorbing 30-60 minute, filtrate is made for continues to separate the residual whey liquid second time that uses, polymeric adsorbent NaCl eluant solution, eluent is with the milipore filter ultrafiltration or the electrodialysis desalination of 5000 dalton's molecular cut offs, vacuum concentrates, freeze drying gets big molecule glycopeptide powder;
(2) separation of residual whey liquid has two kinds of technologies for the second time:
(2a) residual whey liquid pH is adjusted to 7.5 for the second time, adopt anion exchange resin IRA93 type beta-lactoglobulin adsorbed and ALA, per 100 milliliters of whey liquids use 100-200 gram resin, stirring and adsorbing 30-60 minute, polymeric adsorbent NaCl eluant solution, eluent concentrates with the milipore filter ultrafiltration or the electrodialysis desalination of 10,000 dalton's molecular cut offs, and spraying or freeze drying get beta lactoglobulin and ALA mixture; The pH of filtrate is adjusted to 4.5, and with the milipore filter ultrafiltration and the saturating filter of 100,000 dalton's molecular cut offs, vacuum concentrates, and freeze drying gets immunoglobulin G, and milipore filter filters raffinate for continuing to separate usefulness;
(2b) residual whey liquid ultrafiltration for the second time is concentrated into protein content and reaches 15-20%, regulating the pH value is 3.4-3.6, adopt cationic ion-exchange resin C-150S type adsorbing whey albumen, per 100 milliliters of whey liquids use 140-200 gram resin, stirring and adsorbing 30-60 minute, the resin that leaches takes off albumen and handles raffinate for continuing to separate use, polymeric adsorbent NaCl eluant solution, eluent is with the milipore filter desalination of 10,000 dalton's molecular cut offs, concentrate, get the lactalbumin slurry, spraying or freeze drying get PURE WHEY, again with PURE WHEY or directly be made into the 3-10% lactalbumin aqueous solution with lactalbumin slurry, its pH is adjusted to 3.5-4.5, heating-up temperature 80-100 ℃, gets the lactalbumin coagulant liquid, spray-drying gets the gel PURE WHEY;
(3) milipore filter that (2a) step is produced filter raffinate or (2b) resin that produces of step take off albumen and handle the continuation of raffinate and separate two kinds of technologies are arranged:
(3a) described raffinate is concentrated into contains lactose 30-40%, at 45-50 ℃ with cationic ion-exchange resin PCR553 or PCR883 decationized Y sieve, with anion exchange resin IRA401 or IRA402 exhaustion of yin ion and decolouring, and then with solution concentration to containing lactose 75-80%, with reforming catalyst alkali compounds concentration adjustment in this concentrate is 10-20% (w/v), heating-up temperature 80-100 ℃, reacted 30-60 minute, make the lactose isomerization change into lactulose, be quickly cooled to 50 ℃ with chilled brine, with cationic ion-exchange resin PCR553 or PCR883 or use the electrodialysis desalination cessation reaction, remove by filter unreacted lactose, concentrate Cephulac;
(3b) from described raffinate, separate lactose, be processed into the lactose powder, isolated crystalline mother solution is for continue to separate using;
(4) pH with crystalline mother solution is adjusted to 5.0-5.5, inserts ion exclusion and separates in the chromatographic column, and cationic ion-exchange resin PCR553NH is housed in the post
4Type or anion exchange resin IRA401Cl type or IRA401CO
3Type, mother liquor feed volume are the resin bed volumes of 2-14%, flow velocity be the 0.06-0.24 liter/hour, temperature 60-80 ℃, the water wash-out is at first collected the eluent that contains inorganic salts and lactose fraction, concentrates, drying, the salt powder of must suckling, and then collect the eluent that contains lactose fraction.
2. separating technology as claimed in claim 1 is characterized in that described fresh liquid milk is milk, yak milk, buffalo milk or Goat Milk.
3. separating technology as claimed in claim 1 is characterized in that NaCl eluant solution liquid is the NaCl eluant solution liquid of 0.5-1 mole in step (1) and (2).
4. separating technology as claimed in claim 1 is characterized in that being made into the 3-10% lactalbumin aqueous solution with PURE WHEY or lactalbumin slurry again in (2b) step, and its pH is adjusted to 3.8.
5. separating technology as claimed in claim 1, it is characterized in that at raffinate described in (3a) step with cationic ion-exchange resin PCR553 or PCR883 decationized Y sieve with after with anion exchange resin IRA401 or IRA402 exhaustion of yin ion, further take off residual albumen and polypeptide with anion exchange resin IRA900 or IRA938, and then with solution concentration to containing lactose 75-80%.
6. separating technology as claimed in claim 1 is characterized in that being selected from NaOH, KOH, Ca (OH) at alkali compounds described in (3a) step
2, Na
2CO
3, NaHS or Mg (OH)
2
7. separating technology as claimed in claim 1 is characterized in that the concentration of used chilled brine in (3a) step is 5-10%.
8. separating technology as claimed in claim 1, it is characterized in that in step (4), collecting the eluent that contains lactose fraction integrate with in the step (3b) together that processing makes the lactose powder.
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| CN116982656A (en) * | 2023-08-01 | 2023-11-03 | 安徽天凯生物科技有限公司 | Method and device for separating components in milk and dairy products |
| CN117121946A (en) * | 2023-08-24 | 2023-11-28 | 安徽天凯生物科技有限公司 | Method and device for preparing mother emulsified infant formula milk powder and functional milk powder |
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| US4515823A (en) * | 1982-03-31 | 1985-05-07 | Alfa-Laval Ab | Process for separation of raw milk into cream and skim milk which is pasteurized |
| CN1059640A (en) * | 1990-09-13 | 1992-03-25 | 全苏科学研究所综合利用乳制品部 | The process for subsequent treatment of milk |
| WO2003090545A1 (en) * | 2002-04-24 | 2003-11-06 | Dairy Development Specialists, Inc. | Intermediate ingredient for dairy product manufacturing and method of making the same |
| CN1606917A (en) * | 2003-10-13 | 2005-04-20 | 彭平 | Method for extracting lactalbumin and separating different active ingredient of lactalbumin |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4515823A (en) * | 1982-03-31 | 1985-05-07 | Alfa-Laval Ab | Process for separation of raw milk into cream and skim milk which is pasteurized |
| CN1059640A (en) * | 1990-09-13 | 1992-03-25 | 全苏科学研究所综合利用乳制品部 | The process for subsequent treatment of milk |
| WO2003090545A1 (en) * | 2002-04-24 | 2003-11-06 | Dairy Development Specialists, Inc. | Intermediate ingredient for dairy product manufacturing and method of making the same |
| CN1606917A (en) * | 2003-10-13 | 2005-04-20 | 彭平 | Method for extracting lactalbumin and separating different active ingredient of lactalbumin |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102219846A (en) * | 2011-04-20 | 2011-10-19 | 天津商业大学 | Method for separating CGMP (casein glycomacropeptide) having high content of sialic acid by using ion exchange resin |
| CN102746392A (en) * | 2011-04-22 | 2012-10-24 | 天津科技大学 | Method for preparing high purity Casein Glycomacropeptide by using ion exchange method |
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|---|---|
| CN1817149A (en) | 2006-08-16 |
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