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Extraction and purification method of phosvitin in eggs

A phosvitin and purification method technology, which is applied in the field of food engineering, can solve problems such as limited application, inaccurate detection results, and cumbersome separation methods, and achieve the effects of increased yield, high purity, and simplified production process

Inactive Publication Date: 2013-02-20
TIANJIN UNIV OF COMMERCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although high purity was obtained, its separation method was cumbersome, and the yield was only 0.96% (accounting for dry weight of egg yolk)
In 1994, Losso and Nakai established a simpler salt extraction method based on this, but the process is time-consuming and laborious, and a large amount of organic solvent (n-hexane) is used, which affects the biological safety of the final product
In 2003, Castellani et al. used the property of phosvitin to form precipitates with magnesium ions to extract the protein, and the purity of the obtained protein was only 85%. The subsequent purification used anion high-performance liquid chromatography to obtain high-purity phosvitin, but the Technology is not suitable for scale-up production
Although the literature mentions that its protein content is greater than 85%, its detection method makes the detection result inaccurate: the conversion factor of total nitrogen content is 6.25 in the literature, because phosvitin contains a large amount of phosphorus, therefore, it cannot definite indication of its purity
However, the phosvitin prepared through many experiments according to Ko’s method adopts a more scientific N / P expression method, and its N / P ratio is only 4.44, which still needs further purification, which is not enough to be used in food and medicine to check the purity. demanding industry
In 2012, Canadian scholar Bo et al. directly used anion exchange chromatography to purify phosvitin, and obtained a very high purity (N / P ratio of 2.5) but the recovery rate was only 35.4%. In addition, the expensive and efficient Liquid chromatography limits its application in the food industry

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Separation of phosvitin:

[0027] Add 8 fresh egg yolks (112g in total) with 2 times the volume of deionized water at 4°C, stir at a low speed for 1 hour at 4°C, then centrifuge at 4000×g and 4°C for 30 minutes, discard the supernatant and take the precipitate to obtain crude Production of vitellin.

[0028] Add 4 times the volume (w / v) of 80% ethanol to the obtained precipitate, homogenize it at 3000rpm for 2min, then centrifuge at 4000×g, 4°C for 10min, take the precipitate and repeat the above degreasing process for 3 times, and the obtained precipitate is degreasing of egg yolk lipoprotein.

[0029] The obtained precipitate was leached with 9 times the volume of 10% NaCl solution, and the pH value was adjusted to 4.0 with HCl solution, stirred at 4 °C for 30 min, then centrifuged at 4000 × g, 4 °C for 30 min, discarded the precipitate and took the supernatant. The supernatant was then dialyzed and freeze-dried to obtain the crude phosvitin (PV i ) 2.335g, util...

Embodiment 2

[0034] 1. Isolation of phosvitin.

[0035] 5 fresh egg yolks (total 80g), add 3 times the volume of deionized water at 10°C, stir at 10°C at low speed for 30min, then centrifuge at 4500×g, 10°C for 20min, discard the supernatant and take the precipitate to obtain Crude vitellin.

[0036] Add 5 times the volume (w / v) of 85% ethanol to the obtained precipitate, homogenize it at 4000rpm for 1min, then centrifuge at 4500×g, 10°C for 10min, take the precipitate and repeat the above degreasing process twice, and the obtained precipitate is after degreasing of egg yolk lipoprotein.

[0037] The resulting degreased precipitate was washed with 10 times the volume of 9% NH 4 SO 4 The solution was leached, and the pH value was adjusted to 5.0 with HCl solution, stirred at 10°C for 1 hour, and then centrifuged at 4500×g and 10°C for 20 minutes, discarding the precipitate and taking the supernatant. The supernatant was dialyzed and freeze-dried to obtain 1.998g of phosvitin crude produ...

Embodiment 3

[0042] 1. Isolation of phosvitin.

[0043] 7 fresh egg yolks (103g in total), add 1 times the volume of deionized water at 4°C, stir at 4°C at low speed for 1.5h, then centrifuge at 5000×g, 8°C for 10min, discard the supernatant to get the precipitate Crude vitellin is obtained.

[0044] Add 3 times the volume (w / v) of 75% ethanol to the obtained precipitate, homogenize it at 5000rpm for 1min, then centrifuge at 5000×g, 8°C for 10min, take the precipitate and repeat the above degreasing process once, and the obtained precipitate is after degreasing of egg yolk lipoprotein.

[0045] The resulting defatted precipitate was leached with 11 times the volume of 12% NaCl solution, and the pH value was adjusted to 6.0 with HCl solution, stirred at 8 °C for 10 min, and then centrifuged at 5000 × g at 8 °C for 10 min, discarding the precipitate and taking the supernatant. The supernatant was then dialyzed and freeze-dried to obtain the crude phosvitin (PV i ) 2.459g, utilize the Kjel...

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Abstract

The invention discloses an extraction and purification method of phosvitin in eggs, which can be directly applied to foods, drugs and other industries with higher requirement on purity, produces products with high purity, is easy to operate and produce industrially. The method comprises the following steps of: removing water-soluble substances from yolk in an egg; degreasing with ethanol; extracting with high-concentration salt liquid to obtain a phosvitin coarse product; purifying the phosvitin coarse product by utilizing an anion exchange chromatography, and removing salt to obtain high-purity phosvitin. According to the method, the phosvitin is purified by adopting the anion exchange chromatography, so that the purity of the phosvitin can be greatly improved, and the reclamation of the phosvitin is high. Each step in the process is easy to operate, the equipment is low in cost, industrial production can be realized easily, and the phosvitin can be directly applied to the industry of foods and drugs.

Description

technical field [0001] The invention relates to the technical field of food engineering, in particular to a method for extracting and purifying phosvitin from poultry eggs. Background technique [0002] PVS, or phosvitin, is the main phosphoprotein in egg yolk, accounting for about 7% of egg yolk protein. There are two types of α-PVS (37, 42 and 45Ku) and β-PVS (45Ku). - Many PVS, phosvitin has unique physical and chemical characteristics, the phosphorus content in its molecule is about 10%, which is the highest phosphorylation degree among known proteins, the main reason is that about 50% of the amino acid residues in its molecule are serine residues, and 90% of serine residues are phosphorylated, it is because of its highly phosphorylated properties that it has strong emulsifying properties, antioxidant properties, bactericidal properties and metal ion chelating power (about 95% of the serine residues contained in egg yolk % of iron ions combined with it), can be used in ...

Claims

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Application Information

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IPC IPC(8): C07K14/465C07K1/18
Inventor 吴子健王亮吕瑜峰李建颖孙佩佩
Owner TIANJIN UNIV OF COMMERCE
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