CN105348379A - Method for extracting beta-lactoglobulin from milk - Google Patents
Method for extracting beta-lactoglobulin from milk Download PDFInfo
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- CN105348379A CN105348379A CN201510940451.8A CN201510940451A CN105348379A CN 105348379 A CN105348379 A CN 105348379A CN 201510940451 A CN201510940451 A CN 201510940451A CN 105348379 A CN105348379 A CN 105348379A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4717—Plasma globulins, lactoglobulin
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Abstract
The invention discloses a method for extracting beta-lactoglobulin from milk, which comprises the following steps: 1) carrying out first centrifugation on fresh milk so as to obtain skim milk; 2) carrying out second centrifugation on the skim milk so as to obtain whey, and carrying out ultrafiltration on the whey so as to obtain a first permeate; 3) adding a mixture of retinal and ethyl acetate into the first permeate, and carrying out filter pressing on the obtained mixture by using a first plate-and-frame filter press so as to obtain a first retentate; 4) dissolving the first retentate in a sodium hydroxide solution, and carrying out filter pressing on the obtained object by using a second plate-and-frame filter press so as to obtain a second retentate, adjusting the pH value of the second retentate by using hydrochloric acid, so that the pH value of the second retentate is 4-6, and passing the second retentate through a nanofiltration membrane under the action of second pressure, so that a third retentate is obtained; and 5) rinsing and drying the third retentate, so that the beta-lactoglobulin is obtained. The method disclosed by the invention solves the technical problem that the beta-lactoglobulin is not easily extracted.
Description
Technical field
The invention belongs to the production technical field of milk-product, particularly a kind of method extracting beta-lactoglobulin from milk.
Background technology
The many advantages such as whey-protein adopts advanced technologies from milk separation and Extraction precious protein out, high with its purity, specific absorption is high, amino acid composition is the most reasonable are pushed away as " king of albumen ".Whey-protein is not only readily digested, but also has high biological value, high digestibility, high protein effect ratio and high utilization rate, is the features such as the fine work in protein, is one of generally acknowledged human body high-quality protein supplement.In the composition of milk, 87% is water, and 13% is milk solids.And 27% be milk protein in milk solids, only have 20% to be whey-protein in milk protein, all the other 80% are all caseins, and therefore the content of whey-protein in milk is only 0.7%.
Isolated in China, prepare before whey isolate protein starts from more than ten years, the technology adopted at that time mainly membrane technique.And the U.S. just adopts ion exchange method sepg whey albumen from whey as far back as nineteen ninety.In addition, bioselection absorption, affinity chromatography method of purification and reversed phase micelle extraction process etc. are all the methods of the new sepg whey protein ingredient of exploitation.
Current China does not still have the whey-protein comprehensive utilization technique of independent intellectual property right, and foreign technology is by patent protection, and this seriously constrains the exploitation of China's whey-protein resource and the Sustainable development of dairy industry.It is reported, China still needs the whey products such as a large amount of desalted whey powder of import, whey protein concentrate, whey isolate protein and lactose to produce and icecream mix for formula food, candy every year, and its cost is huge.
Develop in external dairy industry and used ultra-filtration technique to make " pre-cheese " and whey enriched material.Compared with traditional evaporation concentration, membrane technique can not only reduce the protein denaturation heating and cause, and has clear superiority in product purification.Be dried to powder at the whey Direct spraying of the U.S. about 2/3rds or make enriched material, the whey that its excess-three is divided does further process, be processed into other whey products, as desalted whey powder, low-lactose whey powder, high lactose whey powder, lactose, whey protein concentrate and whey isolate protein etc.
As can be seen here, also not easily extract whey-protein from from milk at present, extraction efficiency is not high, particularly not easily isolates specific native whey protein composition, and as beta-lactoglobulin, or extraction cost is large, cannot industrial application.
Summary of the invention
For above-mentioned technical problem, propose a kind of method extracting beta-lactoglobulin from milk in the present invention, by twice sheet frame, beta-lactoglobulin is flocculated out, by adding, hot acid is heavy to be extracted beta-lactoglobulin again, solves the technical problem that beta-lactoglobulin not easily extracts.
In order to realize, according to these objects of the present invention and other advantage, providing a kind of method extracting beta-lactoglobulin from milk, comprise the following steps:
Step 1) fresh milk is carried out first time centrifugally operated, remove upper-layer fat, obtain skimmed milk;
Step 2) skimmed milk is carried out second time centrifugally operated, extract supernatant liquor, obtain whey; After whey is carried out pasteurize, at a first pressure by ultra-filtration membrane, obtain the first penetrant after by described ultra-filtration membrane;
Step 3) described first permeate solution is heated to 40 ~ 50 degrees Celsius after, add the mixture of retinene and ethyl acetate, and constantly stir, after being down to normal temperature, by the first plate-and-frame filter press press filtration, obtain the first retentate;
Step 4) the first retentate is dissolved in the solution of sodium hydroxide, and by the second plate-and-frame filter press press filtration, obtain the second penetrant, the pH value of the second penetrant is regulated with hydrochloric acid, make the pH value of the second permeate solution between 4 ~ 6, then the second penetrant is passed through nanofiltration membrane under the second pressure, obtain the 3rd penetrant;
Step 5) the 3rd penetrant rinsing is dry, obtain beta-lactoglobulin.
Preferably, described first time the rotating speed of centrifugally operated be 4000r/min ~ 5500r/min, centrifugal 15min.
Preferably, the rotating speed of described second time centrifugally operated is 5000r/min ~ 6500r/min, centrifugal 20min.
Preferably, described step 2) in, obtain described whey after the 12 ~ 24h that dialysed between 3 ~ 8 degrees Celsius by supernatant liquor.
Preferably, described first pressure is 0.18MPa ~ 0.25MPa.
Preferably, described second pressure is 0.20MPa ~ 0.25MPa.
Preferably, described step 2) in, after whey being heated to 35 ~ 45 degrees Celsius, then by described ultra-filtration membrane.
Preferably, described step 4) in, after the second penetrant is heated to 30 ~ 45 degrees Celsius, then by described nanofiltration membrane.
Preferably, described step 2) in, by supernatant liquor by activated carbon column, then carry out pasteurize.
The present invention at least comprises following beneficial effect:
1, extracting method of the present invention, by twice sheet frame, reheat acid heavy extraction beta-lactoglobulin afterwards, extraction yield is high, reduces tooling cost;
2, extracting method of the present invention can not introduce other foreign ions in the process extracting beta-lactoglobulin, further increases the purity of product.
Part is embodied by explanation below by other advantage of the present invention, target and feature, part also will by research and practice of the present invention by those skilled in the art is understood.
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet extracting beta-lactoglobulin.
Embodiment
The present invention is described in further detail below, can implement according to this with reference to specification sheets word to make those skilled in the art.
Should be appreciated that used in the present inventionly such as " to have ", other element one or more do not allotted in " comprising " and " comprising " term or the existence of its combination or interpolation.
To achieve these goals, the present invention is by the following technical solutions: a kind of method extracting beta-lactoglobulin from milk, shown in Fig. 1, comprises the following steps:
Step 1) fresh milk is carried out first time centrifugally operated on whizzer, remove upper-layer fat, obtain skimmed milk, described first time, the rotating speed of centrifugally operated was 4000r/min ~ 5500r/min, and in the present embodiment, the rotating speed of whizzer adopts 4500r/min, after centrifugal 15min, after leaving standstill for some time, upper-layer fat, in the superiors, extracts by the fat in milk;
Step 2) milk after degreasing is carried out second time centrifugally operated on whizzer, the rotating speed of described second time centrifugally operated is 5000r/min ~ 6500r/min, in the present embodiment, the rotating speed of whizzer adopts 6000r/min, centrifugal 20min and leaving standstill after the time of one end, extract supernatant liquor, obtain whey; Subsequently, after whey is carried out pasteurize, at a first pressure by ultra-filtration membrane, carry out ultrafiltration, described first pressure is 0.18MPa ~ 0.25MPa, and in the present embodiment, the first pressure is 0.19MPa, after ultrafiltration, obtains the first penetrant after by described ultra-filtration membrane;
Step 3) described first permeate solution is heated to 40 ~ 50 degrees Celsius after, add the mixture of retinene and ethyl acetate, the addition of mixture is 0.25 ~ 0.45mg/ml, in the present embodiment, the addition of the mixture of retinene and ethyl acetate is 0.25mg/ml, adds rear and constantly stirs, after during stirring 2h, and after solution to be mixed is down to normal temperature, by mixing solutions by the first plate-and-frame filter press press filtration, being trapped in, filter screen is the first retentate;
Step 4) the first retentate rinsed and is dissolved in the solution of sodium hydroxide, and by the second plate-and-frame filter press press filtration, through filter screen is the second penetrant, the pH value of the second penetrant is regulated afterwards with hydrochloric acid, make the pH value of the second permeate solution between 4 ~ 6, in the present embodiment, pH value is adjusted to 4.8, then the second permeate solution is passed through nanofiltration membrane under the second pressure, obtain the 3rd penetrant through nanofiltration membrane; Described second pressure is 0.20MPa ~ 0.25MPa, and in the present embodiment, the second pressure is 0.22MPa;
Step 5) the 3rd permeate solution rinsing is dry, just obtain beta-lactoglobulin.
In technique scheme, described step 2) in also comprise: obtain described whey, such as 18h after the supernatant liquor of extraction being placed in the environment between 3 ~ 8 degrees Celsius the 12 ~ 24h that dialyses.
In technique scheme, described step 2) in, after whey being heated to 35 ~ 45 degrees Celsius, again by described ultra-filtration membrane, in the present embodiment, whey is heated to 45 degrees Celsius, carry out other compositions in maximum filtering whey with this, reach the accounting improving beta-lactoglobulin in whey.
In technique scheme, described step 4) in, after second permeate solution is heated to 30 ~ 45 degrees Celsius, again by described nanofiltration membrane, in the present embodiment, the second permeate solution is heated to 45 degrees Celsius, other compositions in filtering whey are carried out with this, particularly lactose, to reach the accounting of beta-lactoglobulin in raising second permeate solution, and then the purity of the beta-lactoglobulin extracted.
In technique scheme, described step 2) in, by supernatant liquor by activated carbon column, then carry out pasteurize, supernatant liquor is the transmittance in order to improve supernatant liquor by activated carbon column, with other compositions in filtering supernatant liquor.
From the above mentioned, the method extracting beta-lactoglobulin from milk of the present invention, by twice sheet frame, reheat acid heavy extraction beta-lactoglobulin afterwards, extraction yield is high, reduces tooling cost; Further, extracting method of the present invention can not introduce other foreign ions in the process extracting beta-lactoglobulin, improves the purity of product.
Although embodiment of the present invention are open as above, but it is not restricted to listed in specification sheets and embodiment utilization, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other amendment, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the legend described.
Claims (9)
1. from milk, extract a method for beta-lactoglobulin, it is characterized in that, comprise the following steps:
Step 1) fresh milk is carried out first time centrifugally operated, remove upper-layer fat, obtain skimmed milk;
Step 2) skimmed milk is carried out second time centrifugally operated, extract supernatant liquor, obtain whey; After whey is carried out pasteurize, at a first pressure by ultra-filtration membrane, obtain the first penetrant after by described ultra-filtration membrane;
Step 3) described first permeate solution is heated to 40 ~ 50 degrees Celsius after, add the mixture of retinene and ethyl acetate, and constantly stir, after being down to normal temperature, by the first plate-and-frame filter press press filtration, obtain the first retentate;
Step 4) the first retentate is dissolved in the solution of sodium hydroxide, and by the second plate-and-frame filter press press filtration, obtain the second penetrant, the pH value of the second penetrant is regulated with hydrochloric acid, make the pH value of the second permeate solution between 4 ~ 6, then the second penetrant is passed through nanofiltration membrane under the second pressure, obtain the 3rd penetrant;
Step 5) the 3rd penetrant rinsing is dry, obtain beta-lactoglobulin.
2. the method extracting beta-lactoglobulin from milk according to claim 1, is characterized in that, described first time the rotating speed of centrifugally operated be 4000r/min ~ 5500r/min, centrifugal 15min.
3. the method extracting beta-lactoglobulin from milk according to claim 2, is characterized in that, the rotating speed of described second time centrifugally operated is 5000r/min ~ 6500r/min, centrifugal 20min.
4. the method extracting beta-lactoglobulin from milk according to claim 3, is characterized in that, described step 2) in, obtain described whey after the 12 ~ 24h that dialysed between 3 ~ 8 degrees Celsius by supernatant liquor.
5. from milk, extract the method for beta-lactoglobulin as claimed in claim 4, it is characterized in that, described first pressure is 0.18MPa ~ 0.25MPa.
6. from milk, extract the method for beta-lactoglobulin as claimed in claim 5, it is characterized in that, described second pressure is 0.20MPa ~ 0.25MPa.
7. from milk, extract the method for beta-lactoglobulin as claimed in claim 6, it is characterized in that, described step 2) in, after whey being heated to 35 ~ 45 degrees Celsius, then by described ultra-filtration membrane.
8. from milk, extract the method for beta-lactoglobulin as claimed in claim 7, it is characterized in that, described step 4) in, after the second penetrant is heated to 30 ~ 45 degrees Celsius, then by described nanofiltration membrane.
9. from milk, extract the method for beta-lactoglobulin as claimed in claim 8, it is characterized in that, described step 2) in, by supernatant liquor by activated carbon column, then carry out pasteurize.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510940451.8A CN105348379A (en) | 2015-12-16 | 2015-12-16 | Method for extracting beta-lactoglobulin from milk |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510940451.8A CN105348379A (en) | 2015-12-16 | 2015-12-16 | Method for extracting beta-lactoglobulin from milk |
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| CN105348379A true CN105348379A (en) | 2016-02-24 |
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| CN201510940451.8A Pending CN105348379A (en) | 2015-12-16 | 2015-12-16 | Method for extracting beta-lactoglobulin from milk |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105785047A (en) * | 2016-04-18 | 2016-07-20 | 南昌大学 | Method for quantitative detection of allergen beta-lactoglobulin and sensitization residues thereof based on IgE linear epitope polyclonal antibody |
| CN106720455A (en) * | 2016-11-11 | 2017-05-31 | 光明乳业股份有限公司 | A kind of whey, its preparation method and whey powder base-material |
| CN109805092A (en) * | 2018-12-29 | 2019-05-28 | 华南协同创新研究院 | A kind of milk beverage and preparation method thereof inhibiting LDL oxidation |
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Patent Citations (8)
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| CN1606917A (en) * | 2003-10-13 | 2005-04-20 | 彭平 | Method for extracting lactalbumin and separating different active ingredient of lactalbumin |
| CN101652073A (en) * | 2006-12-18 | 2010-02-17 | 精选牛奶场公司 | Dairy compositions and method of making |
| CN102595915A (en) * | 2009-10-28 | 2012-07-18 | 维利奥有限公司 | Whey protein product and a method for its preparation |
| CN103491790A (en) * | 2011-02-18 | 2014-01-01 | 维利奥有限公司 | Milk-based product and a method for its preparation |
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| CN103524612A (en) * | 2013-09-30 | 2014-01-22 | 暨南大学 | Method for separating milk-derived whey proteins in PEG/phosphate aqueous two-phase system |
| WO2015059248A1 (en) * | 2013-10-23 | 2015-04-30 | Arla Foods Amba | High protein denatured whey protein composition, related products, method of production and uses thereof |
| CN104256053A (en) * | 2014-10-11 | 2015-01-07 | 光明乳业股份有限公司 | Preparation method of alpha-lactalbumin powder and/or beta-lactoglobulin powder and product |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105785047A (en) * | 2016-04-18 | 2016-07-20 | 南昌大学 | Method for quantitative detection of allergen beta-lactoglobulin and sensitization residues thereof based on IgE linear epitope polyclonal antibody |
| CN106720455A (en) * | 2016-11-11 | 2017-05-31 | 光明乳业股份有限公司 | A kind of whey, its preparation method and whey powder base-material |
| CN109805092A (en) * | 2018-12-29 | 2019-05-28 | 华南协同创新研究院 | A kind of milk beverage and preparation method thereof inhibiting LDL oxidation |
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