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Phosvitin phosphopeptide enrichment method

A technology of phosvitin and phosphopeptide, which is applied in the field of extraction, purification and separation, can solve the problems of poor stability of magnetic nanoparticles, large amount of organic reagents, complicated operation, etc., and achieves short separation and purification cycle, high adsorption efficiency and simple operation Effect

Active Publication Date: 2015-11-04
江苏中农鹏源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the calcium alcohol precipitation method is simple, it uses a large amount of organic reagents and has certain pollution to the environment; the membrane separation method has serious membrane pollution and requires cleaning and maintenance; the ion exchange resin method needs to be treated before use and during regeneration, and the operation is complicated
It is also reported that the magnetic nanoparticle enrichment method is used for purification, but the magnetic nanoparticles are not only poor in stability (adsorption cannot occur after repeated use three times), but also the imidazole solvent used in the elution process does not meet the application in the food field.

Method used

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  • Phosvitin phosphopeptide enrichment method
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  • Phosvitin phosphopeptide enrichment method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1: egg yolk protein powder partial dephosphorization method

[0053] Egg yolk protein powder is placed in NaOH solutions of different concentrations of 0.05-0.2mol / L, stirred at 37°C for 3 hours to carry out alkaline dephosphorization treatment, and the treated solution is subjected to ultrafiltration to remove phosphate ions (ultrafiltration membrane is 3000K, the return port pressure is 5-10psi, and the inlet pressure is 20-30psi), the dephosphorization rate of egg yolk protein powder treated with different concentrations of NaOH solution was compared.

[0054] The result shows that 0.05mol / LNaOH, dephosphorization rate is 27%; 0.1mol / LNaOH, dephosphorization rate is 46%; when 0.15mol / L, dephosphorization rate is 53%; %.

Embodiment 2

[0055] Example 2: Enzymolysis of partially dephosphorylated egg yolk protein powder

[0056] Enzymatic hydrolysis of partially dephosphorylated egg yolk protein powder found that the dephosphorization rate was low, resulting in a low degree of hydrolysis. However, if the dephosphorylation is excessive, the calcium-storing function of the resulting PPPs is reduced. Therefore, the dephosphorized egg yolk protein powder with a dephosphorization rate of 46%-64% is selected for enzymatic hydrolysis.

[0057] The enzymolysis process is as follows: add dephosphorylated egg yolk protein powder and deionized water into the enzymolysis reactor (substrate concentration is 1%), heat in a water bath at 50°C, and adjust the pH to 8.0 after the solution is evenly stirred. The trypsin was added to the above solution (enzyme:substrate=5:100), titrated with 0.1mol / L NaOH solution during the reaction to maintain the pH at 8.0, and the reaction time was 4h. After the enzymolysis, the above reac...

Embodiment 3

[0058] Example 3: Mesoporous / Macroporous TiO 2 material preparation

[0059] Convenient and rapid synthesis of mesoporous / macroporous TiO by sol-gel method using tetra-n-butyl titanate as titanium source 2 Material.

[0060] The specific experimental method is as follows: First, weigh 0.3g of citric acid, dissolve it in 40mL of 95% absolute ethanol, add 18mL of ammonia solution after magnetic stirring for 30min, continue stirring for 10min, and then take 5mL of tetra-n-butyl titanate , under stirring, the above solution was added dropwise, and after magnetic stirring for 1 h, it was left to stand at room temperature for 12 h. Then, the precipitate was washed successively with distilled water and absolute ethanol, and the washed material was dried in a drying oven at 60° C. for 14 h. Finally, the above materials were calcined in a temperature-programmed muffle furnace at 550° C. for 4 h.

[0061] The resulting mesoporous / macroporous TiO 2 The scanning electron microscope i...

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Abstract

The invention discloses a phosvitin phosphopeptide enrichment method and belongs to the field of extraction, purification and separation. The phosvitin phosphopeptide enrichment method realizes enrichment of phosvitin phosphopeptide by mesoporous / macroporous TiO2 as an adsorption material. The prepared phosvitin phosphopeptide has N / P of 4.5. Mesoporous / macroporous TiO2 can utilize metal oxide TiO2 specific affinity effects on a phosphate group. Through high specific surface area and a good tunnel structure of the mesoporous-macroporous structure, a large amount of active binding sites are provided. Under enrichment conditions of pH of 2.5-4, an initial polypeptide concentration of 12-20mg / mL and adsorption time of 5-20min, the PPPs has purity of 4.3-4.8 and the corresponding adsorption quantity is in a range of 42-53mg / mL. The phosvitin phosphopeptide enrichment method has simple processes, is fast and effective and is conducive to industrial production.

Description

technical field [0001] The invention relates to a method for enriching phosvitin phosphopeptide, which belongs to the field of extraction, purification and separation. Background technique [0002] Egg yolk protein powder is a by-product of the industrial production of egg yolk lecithin. More and more scholars have conducted comprehensive research on it and made some progress. Phosvitin (Phosvitin, PV) is the main phosphorus-containing protein in defatted egg yolk protein powder. Its primary structure contains 217 amino acid residues, of which 124 are serine residues, accounting for about 56% of the total amino acid residue content. . Numerous studies have shown that phosvitin can interact with Ca 2+ , Mg 2+ , Mn 2+ ,Co 2+ ,Sr 2+ , Fe 2+ , Fe 3+ Wait for the combination. Since phosvitin is firmly combined with metal ions and is not easily hydrolyzed, its biological potency is low, which is not conducive to nutrient absorption. It has been confirmed that the main fu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/14C07K14/465A23L1/305
Inventor 杨严俊苏宇杰李松娜
Owner 江苏中农鹏源生物科技有限公司
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