Method for extracting osteopontin in cow's milk

A technology of osteopontin and extraction method, applied in the direction of animal protein processing, etc., can solve the problem of low purity of osteopontin, and achieve the effects of short extraction time, simple operation and low cost

Inactive Publication Date: 2009-07-22
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for extracting osteopontin ...

Method used

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specific Embodiment approach 1

[0008] Specific Embodiment 1: The extraction method of osteopontin in milk according to the present embodiment is realized by the following steps: 1. Ion-exchange chromatographic separation: 1000-2000mL milk is centrifuged for 15-30min, and after removing the sediment, mix with 200-400mL DEAE-Sephacel resin , stirred at 3-6°C for 12-24 hours, then stood still, poured into the chromatographic column after removing the precipitate, and then used the concentration of 0.01-0.015mol / L and pH value under the condition of flow rate of 3-5mL / min Equilibrate the chromatographic column with a phosphate buffer solution of 7.2-7.6, and then carry out gradient elution under the conditions of a flow rate of 3-5mL / min and a detection wavelength of 280nm. The gradient elution is completed in three stages, and the samples of each stage are collected separately. Gradient eluent; 2. Use hydrophobic chromatography to separate the gradient elution obtained in step 1, respectively, to obtain eluents...

specific Embodiment approach 2

[0009] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the eluent in the first stage of gradient elution in step 1 is that the concentration of NaCl is 0.2 mol / L, the pH value is 7.0-7.2, and the concentration is 0.01-0.015 mol / L phosphate buffer solution; gradient elution The second stage eluent is a phosphate buffer solution with a NaCl concentration of 0.25mol / L, a pH value of 7.0-7.2, and a concentration of 0.01-0.015mol / L; gradient elution The eluent in the third stage is a phosphate buffer solution with a NaCl concentration of 0.3 mol / L, a pH value of 7.0-7.2, and a concentration of 0.01-0.015 mol / L. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0010] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is that the hydrophobic chromatographic separation in step two is carried out according to the following steps: add NaCl to each gradient eluent in the gradient eluent obtained in step one respectively The concentration of NaCl is 4mol / L, and then equilibrate the chromatographic column with a phosphate buffer solution with a concentration of 0.01-0.015mol / L and a pH value of 7.2-7.6 under the condition of a flow rate of 1-3mL / min. Under the conditions of ~3mL / min and detection wavelength of 280nm, phosphate buffer solution with NaCl concentration of 2mol / L, pH value of 7.0~7.2, and concentration of 0.01~0.015mol / L was used as eluent for elution respectively. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a method for extracting osteopontin in cow milk, relates to a method for extracting osteopontin, and solves the problem that the extracted osteopontin in the cow milk has low purity in the prior art. The extracting method comprises the following steps of ion-exchange chromatographic separation, primary hydrophobic chromatographic separation, secondary hydrophobic chromatographic separation, dialysis, and freeze-drying to prepare the osteopontin in the cow milk. The extracted osteopontin in the cow milk has high purity. The method does not adopt a step of protein denaturation, and has the advantages of simple and convenient operation, low cost, and short extraction time.

Description

technical field [0001] The invention relates to a method for extracting osteopontin. Background technique [0002] Milk contains almost all the nutrients needed by the human body and is the only complete nutritional food. Milk is easy to digest and absorb, high in quality and low in price, convenient to eat, and contains many biologically active substances. It is an excellent source of high-quality protein in human food. Osteopontin (OPN) is a highly phosphorylated glycoprotein rich in sialic acid, mainly distributed in mineral tissues (such as bone and cementum) as a non-collagenous extracellular matrix protein, and promotes osteoclastosis Adhesion of cells and promotion of osteoid biomineralization. Several studies have shown that osteopontin is present in bone cells, adheres to the extracellular matrix, and can act as a chemical inducer in the early stages of bone formation. At present, the research on osteopontin and bone health has attracted more and more people's at...

Claims

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Application Information

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IPC IPC(8): A23J3/08
Inventor 刘宁孙婕
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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