CN1849903A - Method for inductrialized separation of purified lactoferrins from cattle colostrum - Google Patents
Method for inductrialized separation of purified lactoferrins from cattle colostrum Download PDFInfo
- Publication number
- CN1849903A CN1849903A CN 200610040528 CN200610040528A CN1849903A CN 1849903 A CN1849903 A CN 1849903A CN 200610040528 CN200610040528 CN 200610040528 CN 200610040528 A CN200610040528 A CN 200610040528A CN 1849903 A CN1849903 A CN 1849903A
- Authority
- CN
- China
- Prior art keywords
- colostrum
- lactoferrin
- ultrafiltration
- enzyme
- separation
- 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.)
- Granted
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention relates to a method for industrial separation and purification of lactoferrin from beestings. It is characterized by that it uses beestings defatted serum as raw material, adopts enzyme method to remove casein and utilizes combination of membrane separation technique and industrial chromatographic technique to separate out lactoferrin whose purity is above 90% from beestings.
Description
Technical field
A kind of from colostrum the method for inductrialized separation of purified lactoferrins, relate to the application of modern biological integrated technology in deep processing of farm products such as membrane separation technique and industry chromatography technique, belong to industrial separation and purification of lactoferrin in the colostrum.
Background technology
Dairy industry circle it is generally acknowledged that cow divides the milk in 7 days puerperiums to be called colostrum.Colostrum is a kind of very special breast, and its chemical composition and processing characteristics and common milk have very big difference.The colostrum color and luster is yellow and dense thick, even has blood redly on a small quantity, has special newborn fishy smell.The milk solids content of colostrum is higher than normal breast, and wherein globulin, albumin and inorganic salt content are high especially.The acidity of colostrum is very high, poor heat stability.Just a large amount of antibiotic, agricultural chemicals and synthetic auxin also may be contained in the Ruzhong.Because these special natures, colostrum belongs to " abnormal milk ", U.S. food and FAD (FDA) clearly regulation colostrum must not mix normal breast processing, various countries' quality of dairy products standard general provision forbids to use colostrum as raw milk, most pastures all with colostrum as calf feed or discarded.
Yet colostrum contains the material of multiple biologically active function, as immunoglobulin (Ig), lactoferrin, lysozyme, active peptide, lactoperoxidase, growth factor etc., all has special bioactive functions.Though these content of active substance are a little less than people's colostrum, far above other food in the colostrum.
In recent years, the lactoferrin in first Ruzhong is subjected to the extensive concern of Chinese scholars.It is a kind of glycoprotein of biologically active function, has multiple bioactive functions, transmission and absorption, immunoregulation effect, adjusting inflammatory response, antivirus action and the effect of inhibition fat oxidation as broad spectrum antibiotic activity, enhancing iron can be widely used in food, medicine and cosmetic field.
The method of separating lactoferrin mainly contains ion-exchange chromatography, metal chelate chromatography, hydrophobic chromatography, affinity chromatography, immunochromatography and ultrafiltration.The lactoferrin purity that ion-exchange chromatography, metal chelate chromatography, hydrophobic chromatography separation obtain is higher; But adsorption capacity is little, efficient is low.Affinity chromatography can be realized step separation, and electrophoretic analysis shows its based on very high purity, good separating effect, and biologically active is loss not; But cost is higher, and the industrialization difficulty is bigger.It is reusable after immunochromatography good separating effect, purity height, antibody are fixed; But the complicated process of preparation of post, antibody cost costliness is difficult to realize industrialization.The Treatment with Ultrafiltration amount is big, but the purity of product is lower.Generally believe that both at home and abroad affinity chromatography is one of effective method of production pharmaceutical grade lactoferrin, ultrafiltration is used to the with the largest potentiality of production food-grade lactoferrin.
This shows that though some isolation technics can make highly purified lactoferrin in laboratory scale, treating capacity is little, and is with high costs, be difficult to realize industrialization; And ultrafiltration output is big, but purity is low, thereby has hindered applying of lactoferrin.Seeking moderate, the method fast and effectively of a kind of cost at industrially scalable, isolate the higher lactoferrin of purity from colostrum, is dairy husbandry problem demanding prompt solution both at home and abroad at present.
Summary of the invention
The purpose of this invention is to provide a kind of fast effectively, moderate, the high-purity of cost, high-recovery, the method for inductrialized separation of purified lactoferrins from colostrum.
Technical scheme of the present invention: with the colostrum skimmed whey is raw material, adopts enzyme process to remove casein, and membrane separation technique and industry chromatography technique combine, and industrial separation purity is the lactoferrin more than 90% from colostrum.Process route as shown in Figure 1.
1. enzyme process is removed casein: with the endocrine colostrum of 72h is raw material, centrifugal degreasing, adopt rennin enzyme process precipitation casein, the clarity of whey is good, alleviated the coprecipitation phenomena of protein, improved the retention rate of lactoferrin in the whey effectively, this method is better than traditional salt acid precipitation method, can not cause environmental pollution.
In the colostrum skimmed whey, add rennin liquid, the vigor of rennin is 2000/mL, addition is to add this enzyme liquid 50mL in the 100kg colostrum skimmed whey, enzyme concentration is controlled at that colostrum 10-15min is solidified as is suitable, and the pH of reaction system is adjusted to 6.0, and enzyme reaction temperature is 40 ℃, colostrum solidifies back insulation 30min, and the gained supernatant is the enzyme process whey behind the centrifugal removal casein.
2. pre-filtering: gained enzyme process whey by decline filter clarification of capsule, is removed residual junket egg from micelle.Also residual a large amount of casein micelle in the enzyme process whey that process link-suspended basket centrifuge removal casein post precipitation obtains, therefore necessary the enzyme process whey is carried out pre-filtering, can alleviate the terminal ultrafiltration like this stops up, enhance productivity, prolong the service life of milipore filter in the later separation (terminal separation).The capsule that adopts Suzhou branch company of Pore company the to produce filter that declines, the aperture is 0.2 μ m.These seed capsules filter that declines is the filter of the model of instant use once to turn it on that is integrated of a kind of novel filter core and its plastic casing.Not only hold dirty ability and hold back that efficient is strong, percent opening is high, and can be at any time renew cartridge more, very convenient, dead volume is little, therefore in long-term use, can keep high flow rate and high flux.Hold back relation between efficient and the production capacity for balance, reduce the pre-filtering cost to greatest extent, it is optimum adopting the capsule formula to filter.
3. substep ultrafiltration: the ultrafiltration system that adopts plate type membrane component and cellulose acetate membrane material, operates in the cross-flow mode, at first selecting for use and holding back relative molecular mass is that the milipore filter of 150kDa carries out first step ultrafiltration, collect and see through liquid, selecting for use and holding back relative molecular mass is that the milipore filter of 10kDa carries out the second step ultrafiltration, the ultrafiltration technology parameter is: tangential flow velocity 3-4L/min, critical operation pressure 0.10-0.15MPa, operating temperature 45-55 ℃, hold back concentrate and be the lactoferrin semifinished product, the purity of lactoferrin is about 30%.
4. process-scale chromatography purifying: select SP Sepharose Fast Flow strong cation exchanger for use, the dress post, column dimension is φ 7.0 * 20cm, with the pH value is 7.7～8.0, the phosphate of 10mmol/L is initial level pad, hold back sample on the concentrate, elution speed 2L/h, adopt other component of NaCl eluant solution of 0.5mol/L earlier, return back to baseline values to the wash-out absorption value, adopt the NaCl eluant solution lactoferrin component of 1mol/L again, return back to baseline values once more to the wash-out absorption value and finish, collect the eluent of this lactoferrin component as wash-out, through the ultrafiltration desalination, concentrate and freeze drying, obtain the lactoferrin highly finished product.Measure through sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE), gained lactoferrin component is shown as single district band, and relative molecular mass is 80,400Da.The purity and the rate of recovery of lactoferrin are respectively 94.20% and 75.45% in the lactoferrin highly finished product of producing.
Beneficial effect of the present invention: along with the raising of China's living standards of the people and the enhancing of health perception, particularly to the understanding of cow's milk nutritive value, dairy processing industry has obtained swift and violent development, has also caused the increase of first milk production simultaneously.The annual discarded colostrum resource of China is above 180,000 tons.Both at home and abroad breastfeeding benefit is reached common understanding.But breast milk is limited after all, and human nature will be sought other colostrum resource.Take all factors into consideration factors such as ethics, religion, consumption habit and industrial foundation, the functional components that people transfer in the exploitation colostrum is an inexorable trend, because it is the most real up to now colostrum resource.
Lactoferrin is a kind of glycoprotein that is present in the biologically active function in the exocrine secretion (as breast).The content of lactoferrin is the abundantest in first Ruzhong.It has multiple bioactive functions, transmission and absorption, immunoregulation effect, adjusting inflammatory response, antivirus action and the effect of inhibition fat oxidation as broad spectrum antibiotic activity, enhancing iron are acknowledged as a kind of important function composition or the trophic function factor.Lactoferrin mainly can be applicable to following three aspects: 1. field of food is developed as natural anticorrisive agent and antiseptic; Add in the varieties of food items as functional components, for example dairy products, sport nutrition product, instant beverage, immune product and chewing gum etc.2. field of medicaments is made the natural antibacterial medicine or is efficiently mended the iron medicament.3. the physical function of nascent animal is improved as feed addictive and positive growth factor in other field, is used for cosmetics and strengthens antidotal effect.
The large-scale production lactoferrin is the problem that presses for solution in the lactoferrin research.At present, though some isolation technics can make highly purified lactoferrin in laboratory scale, treating capacity is little, and is with high costs, is difficult to realize industrialization, thereby hindered applying of lactoferrin.The present invention is just at above technological difficulties, invented moderate, the method fast and effectively of a kind of cost at industrially scalable, isolates the method for the higher lactoferrin of purity from colostrum.
The present invention improves particularly infant's the general level of the health of China people for the deep processing that promotes dairy products and the comprehensive utilization of waste resource, and the research and development functional food ingredient all has great importance.
Description of drawings
The process flow diagram of Fig. 1 inductrialized separation of purified lactoferrins from colostrum.
The specific embodiment
Embodiment 1
The above-mentioned process separation and purification of by specification lactoferrin.Skimmed colostrum amount: 100L, lactoferrin content 1.50g/L, enzyme process obtains enzyme process whey 90L after removing casein, two step films obtain concentrate 18L after separating, lactoferrin concentration 6.91g/L, the lactoferrin rate of recovery (in first Ruzhong lactoferrin content) 82.92%, lactoferrin is 31.40% with respect to the purity of total protein in the concentrate in the concentrate.Concentrate obtains lactoferrin highly finished product 113.18g after chromatographic isolation, desalination and concentration and freeze drying, purity is 94.20%, and the lactoferrin rate of recovery (in first Ruzhong lactoferrin content) is 71.08%.
Embodiment 2
Operation is with embodiment 1, skimmed colostrum amount: 300L, lactoferrin content 1.48g/L, enzyme process obtains enzyme process whey 272L after removing casein, two step films obtain concentrate 53L after separating, lactoferrin concentration 7.05g/L, the lactoferrin rate of recovery (in first Ruzhong lactoferrin content) 84.16%, lactoferrin is 32.91% with respect to the purity of total protein in the concentrate in the concentrate.Concentrate obtains lactoferrin highly finished product 350.35g after chromatographic isolation, desalination and concentration and freeze drying, purity is 95.92%, and the lactoferrin rate of recovery (in first Ruzhong lactoferrin content) is 75.72%.
Claims (1)
1. the method for an inductrialized separation of purified lactoferrins from colostrum, it is characterized in that with the colostrum skimmed whey be raw material, adopt enzyme process to remove casein, membrane separation technique and industry chromatography technique combine, and industrial separation purity is the lactoferrin more than 90% from colostrum;
(1) enzyme process is removed casein: in the colostrum skimmed whey, add rennin liquid, the vigor of rennin is 2000U/mL, addition is to add this enzyme liquid 50mL in the 100kg colostrum skimmed whey, enzyme concentration is controlled at that colostrum 10-15min is solidified as is suitable, and the pH of reaction system is adjusted to 6.0, and enzyme reaction temperature is 40 ° of C, colostrum solidifies back insulation 30min, and the gained supernatant is the enzyme process whey behind the centrifugal removal casein;
(2) pre-filtering: gained enzyme process whey by decline filter clarification of capsule, is removed residual casein micelle;
(3) substep ultrafiltration: the ultrafiltration system that adopts plate type membrane component and cellulose acetate membrane material, operates in the cross-flow mode, at first selecting for use and holding back relative molecular mass is that the milipore filter of 150kDa carries out first step ultrafiltration, collect and see through liquid, selecting for use and holding back relative molecular mass is that the milipore filter of 10kDa carries out the second step ultrafiltration, the ultrafiltration technology parameter is: tangential flow velocity 3-4L/min, critical operation pressure 0.10-0.15MPa, holds back concentrate and is the lactoferrin semifinished product by operating temperature 45-55 ℃;
(4) process-scale chromatography purifying: select SP Sepharose Fast Flow strong cation exchanger for use, the dress post, column dimension is φ 7.0 * 20cm, with the pH value is 7.7～8.0, the phosphate of 10mmol/L is initial level pad, hold back sample on the concentrate, elution speed 2L/h, adopt other component of NaCl eluant solution of 0.5mol/L earlier, return back to baseline values to the wash-out absorption value, adopt the NaCl eluant solution lactoferrin component of 1mol/L again, return back to baseline values once more to the wash-out absorption value and finish, collect the eluent of this lactoferrin component as wash-out, through the ultrafiltration desalination, concentrate and freeze drying, obtain the lactoferrin highly finished product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100405287A CN100558242C (en) | 2006-05-22 | 2006-05-22 | A kind of from colostrum the method for inductrialized separation of purified lactoferrins |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100405287A CN100558242C (en) | 2006-05-22 | 2006-05-22 | A kind of from colostrum the method for inductrialized separation of purified lactoferrins |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1849903A true CN1849903A (en) | 2006-10-25 |
| CN100558242C CN100558242C (en) | 2009-11-11 |
Family
ID=37131422
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006100405287A Expired - Fee Related CN100558242C (en) | 2006-05-22 | 2006-05-22 | A kind of from colostrum the method for inductrialized separation of purified lactoferrins |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100558242C (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104151423A (en) * | 2014-07-30 | 2014-11-19 | 哈尔滨工业大学 | Lactoferrin preparation method on basis of casein/lactoferrin dynamic adsorption/desorption mechanism |
| CN104151423B (en) * | 2014-07-30 | 2016-11-30 | 哈尔滨工业大学 | A kind of lactoferrin preparation method based on casein Yu lactoferrin dynamic adsorption mechanism for resolving |
| CN106589103A (en) * | 2016-12-21 | 2017-04-26 | 武汉华科大生命科技有限公司 | Preparation method of iron proteinsuccinylate |
| CN107022593A (en) * | 2017-05-12 | 2017-08-08 | 江苏天美健大自然生物工程有限公司 | A kind of Colostrinin of Pro-rich polypeptide and preparation method thereof |
| CN109156592A (en) * | 2018-08-28 | 2019-01-08 | 吉林大学 | A kind of pressed candy product and preparation method thereof rich in lactoferrin |
| CN109287747A (en) * | 2018-08-28 | 2019-02-01 | 珠海经济特区天然药物研究所有限公司 | A kind of preparation method of colostrum element |
| CN110981951A (en) * | 2019-12-26 | 2020-04-10 | 吉林大学 | Preparation method for enriching immunoglobulin and lactoferrin in colostrum |
| CN115598278A (en) * | 2022-12-14 | 2023-01-13 | 黑龙江飞鹤乳业有限公司（Cn） | Quantitative detection method for lactoferrin |
-
2006
- 2006-05-22 CN CNB2006100405287A patent/CN100558242C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104151423A (en) * | 2014-07-30 | 2014-11-19 | 哈尔滨工业大学 | Lactoferrin preparation method on basis of casein/lactoferrin dynamic adsorption/desorption mechanism |
| CN104151423B (en) * | 2014-07-30 | 2016-11-30 | 哈尔滨工业大学 | A kind of lactoferrin preparation method based on casein Yu lactoferrin dynamic adsorption mechanism for resolving |
| CN106589103A (en) * | 2016-12-21 | 2017-04-26 | 武汉华科大生命科技有限公司 | Preparation method of iron proteinsuccinylate |
| CN107022593A (en) * | 2017-05-12 | 2017-08-08 | 江苏天美健大自然生物工程有限公司 | A kind of Colostrinin of Pro-rich polypeptide and preparation method thereof |
| CN109156592A (en) * | 2018-08-28 | 2019-01-08 | 吉林大学 | A kind of pressed candy product and preparation method thereof rich in lactoferrin |
| CN109287747A (en) * | 2018-08-28 | 2019-02-01 | 珠海经济特区天然药物研究所有限公司 | A kind of preparation method of colostrum element |
| CN110981951A (en) * | 2019-12-26 | 2020-04-10 | 吉林大学 | Preparation method for enriching immunoglobulin and lactoferrin in colostrum |
| CN110981951B (en) * | 2019-12-26 | 2023-03-28 | 吉林大学 | Preparation method for enriching immunoglobulin and lactoferrin in colostrum |
| CN115598278A (en) * | 2022-12-14 | 2023-01-13 | 黑龙江飞鹤乳业有限公司（Cn） | Quantitative detection method for lactoferrin |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100558242C (en) | 2009-11-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100558242C (en) | A kind of from colostrum the method for inductrialized separation of purified lactoferrins | |
| EP0253395B2 (en) | A process for producing bovine lactoferrin in high purity | |
| CA2724444C (en) | Process for producing milk fractions rich in secretory immunoglobulins | |
| CN104186916B (en) | The method that lactalbumin powder is prepared in separation from milk surum | |
| NZ535784A (en) | A process of isolating lactoferrin | |
| CN101603038B (en) | Preparation method of lysozyme | |
| CN103709246A (en) | Purification method for lactoferrin and lactoperoxidase | |
| CN109761833B (en) | A kind of isolation and purification method of L-Leu | |
| CN102870952B (en) | Method for preparing whey protein powder (WPC) and lactose powder simultaneously by whey | |
| CN104313099B (en) | A kind of preparation method of the active egg white powder peptide of high anti-oxidation | |
| CN105648008A (en) | Feeding silk antibacterial peptide preparation and preparation method thereof | |
| CN104450639B (en) | A kind of method using ultrafiltration auxiliary aqueous two phase extraction technique extraction lactoperoxidase | |
| CN100358916C (en) | Method for extracting high purity protein from cow milk or soybean waste water | |
| CA2886892A1 (en) | Improved process for purifying lactoferrin from milk and products thereof | |
| CN1129609C (en) | Flocculation charification-ultrafiltration concentration process of producing composite immunoreactive protein | |
| CN1663961A (en) | Technology for separating and purifying lactoferritin from cow colostrum | |
| CN105399816B (en) | Method for separating human lactoferrin from transgenic goat milk | |
| CN100429231C (en) | Method for removing beta-lactoglobulin in milk and extracting high purity immune globulin | |
| CN104304642B (en) | A kind of concentrated type lactalbumin WPC80 and preparation method thereof | |
| CN107721909A (en) | DNJ, flavones, the method and system of polysaccharide are continuously extracted from moraceae plants | |
| EP0595993A1 (en) | Isolation of charged particles from fluids | |
| Stanic et al. | Application of ion exchanger in the separation of whey proteins and lactin from milk whey | |
| CN111073939A (en) | Production process of high-activity small peptide chelated copper | |
| Mulvihill et al. | Production of whey-protein-enriched products | |
| CN1274716C (en) | Method for extracting albumin transferrin with iron supplemented and antibacterial function |
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 | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20091111 Termination date: 20140522 |