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Method for Catalytic Synthesis of Biomimetic Material Hydroxyapatite Using Phosvitin and Its Polypeptide

A phosvitin and hydroxyapatite technology, applied in phosphorus compounds, chemical instruments and methods, inorganic chemistry, etc., can solve the problems of synthesis time limitation, long time, etc. active effect

Active Publication Date: 2016-06-01
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, the aqueous solution reaction method is easy to operate, has mild conditions and low cost, and has been paid more and more attention. However, the calcium hydrogen phosphate precipitate obtained by the reaction needs to go through a crystal phase transformation process to obtain hydroxyapatite. This process takes a long time. For example, in In "A Method for Synthesizing Hydroxyapatite from Alkali Slag", "the stirring reaction time is 1-2 hours, and the aging time is 12-24 hours", and the longer synthesis time limits the application of this method

Method used

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  • Method for Catalytic Synthesis of Biomimetic Material Hydroxyapatite Using Phosvitin and Its Polypeptide
  • Method for Catalytic Synthesis of Biomimetic Material Hydroxyapatite Using Phosvitin and Its Polypeptide

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Embodiment 1

[0024] A method of utilizing phosvitin to catalyze the synthesis of biomimetic material hydroxyapatite:

[0025] After dissolving calcium chloride and potassium dihydrogen phosphate, react in a neutral solution under the catalysis of phosvitin, the molar ratio of calcium chloride and potassium dihydrogen phosphate is 3:1, and add 1 g of calcium ion per 3 mol of calcium ion Phosvitin. During the reaction process, 0.1 mol / L NaOH solution is continuously added dropwise to keep the pH of the system at 6.8-7.2. After 15 minutes of reaction, the precipitate is collected to obtain the hydroxyapatite mineralized material.

Embodiment 2

[0027] 1) Enzymatic hydrolysis of phosvitin: Dissolve phosvitin in 0.01mol / L NaOH solution, stir for 1 hour, adjust the pH of the solution to 2.2, add 1g of pepsin for every 50g of phosvitin, and perform enzymatic hydrolysis for 3 hours , and then adjust the pH of the solution to 7.5, then add 1 g of trypsin for every 20 g of phosvitin, and perform enzymatic hydrolysis for 5 hours. After the enzymatic solution is centrifuged, the supernatant is the phosvitin polypeptide mixture.

[0028] 2) Separation and purification of peptides: using ToyopearlDEAE-650M anion exchange chromatography, the filler equilibration buffer is Tris-HCl buffer with pH 7.2, after the supernatant is loaded, the concentration of sodium chloride is 0, 0.1, 0.2, 0.3 in sequence , 0.4, 0.5, and 0.6 mol / L Tris-HCl buffer solution for stepwise gradient elution, collect the eluate with a sodium chloride concentration of 0.2 mol / L, and perform desalination treatment according to the conventional ultrafiltration ...

Embodiment 3

[0031] (1) Enzymatic hydrolysis of phosvitin: take phosvitin and dissolve it in 0.1mol / L NaOH solution, stir for 0.5 hours, adjust the pH of the solution to 2.8, add 1g of pepsin per 100g of phosvitin, and enzymolyze 5 hour, then adjust the pH of the solution to 8.2, then add 1g of trypsin per 80g of phosvitin, and enzymatically hydrolyze for 8 hours. After the enzymatic solution is centrifuged, the supernatant is the phosvitin polypeptide mixture.

[0032] (2) Separation and purification of peptides: QSepharoseFF anion exchange chromatography is used, the filler equilibrium buffer is Tris-HCl buffer at pH 8.0, after the supernatant is loaded, use Tris-HCl buffer with a sodium chloride concentration of 0.5mol / L The phosvitin polypeptide with biological activity is obtained after the eluate is collected and desalted.

[0033] (3) Catalytic synthesis: react calcium chloride and sodium dihydrogen phosphate under the catalysis of phosvitin polypeptide under neutral conditions, the...

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Abstract

The invention discloses a method for catalytically synthesizing a biomimetic material hydroxyapatite from phosvitin and a polypeptide thereof. The method includes following steps: dissolving a calcium salt and a phosphate, performing a reaction under a neutral condition with catalysis of the phosvitin and the polypeptide thereof, and collecting generated precipitates in the reaction to obtain the biomimetic material hydroxyapatite. In the invention, by means of bioactivities of the phosvitin and athepolypeptide thereof, a crystal phase transition during synthesis of the phosvitin is greatly promoted, a crystal phase transition time is reduced from 16 hours to 5-15 min and a conversion speed is increased by 48-216 times. The phosvitin polypeptide is high in purity and is strong in catalytic activity.

Description

technical field [0001] The invention belongs to the field of bioactive material preparation, and specifically relates to the application of phosvitin and its polypeptide in catalytically synthesizing bionic material hydroxyapatite, and a bionic material hydroxyapatite catalyzed by phosvitin and its polypeptide Methods. Background technique [0002] Phosvitin is a phosphoprotein in egg yolk, containing 10% phosphorus, and is currently the protein with the highest degree of phosphorylation in nature (Anton2007). Phosvitin has many similarities with phosphorylated proteins in the human body (such as osteopontin, dentin matrix protein), and these phosphorylated proteins in the human body have the function of regulating biomineralization (George and Veis2008, Tay and Pashley2009), based on their Due to the similarity in structure, it is speculated that phosvitin and its polypeptides may also have similar functions. Therefore, there is a certain theoretical basis for using phosv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C01B25/32
Inventor 马美湖张晓维耿放黄茜
Owner HUAZHONG AGRI UNIV
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