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Method for rapid purifying soluble protein of shell pearl layer

A protein and nacre technology, applied in the preparation methods of peptides, chemical instruments and methods, animal/human peptides, etc., can solve the problem that soluble shell proteins cannot meet the clinical application of pearls, etc., to promote clinical application, promote bone repair, The effect of speeding up the generation

Inactive Publication Date: 2005-05-11
OCEAN UNIV OF CHINA
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Problems solved by technology

However, the amount of soluble shell protein obtained is far from meeting the clinical application of bone repair in medicine and the needs of in vitro cultured pearls.

Method used

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  • Method for rapid purifying soluble protein of shell pearl layer
  • Method for rapid purifying soluble protein of shell pearl layer
  • Method for rapid purifying soluble protein of shell pearl layer

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Embodiment Construction

[0012] Scrub the surface of the wrinkled abalone shell with tap water, soak it in 5% sodium hydroxide solution for 10 hours, take it out, brush off the surface impurities and attachments, rinse it with distilled water, and dry it in the shade at room temperature for 24 hours to get a clean shell . The shells are treated with acid to remove the cuticle and prismatic layers, resulting in pure nacre. Crush nacre shells with 0.01% NaN 3 The concentration of 5% acetic acid solution was dialyzed for more than 72 hours, and then dialyzed with distilled water for 72 hours to obtain a solution containing nacre soluble shell protein, which was centrifuged at 20,000 g for 30 minutes. Take the supernatant and dry it in vacuum to obtain dry powdery soluble protein.

[0013] The shell protein was separated and purified by fast protein chromatography system (FPLC) and polyacrylamide gel electrophoresis (SDS-PAGE). The specific methods and results are as follows:

[0014] (1) Desalination....

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Abstract

Rapid purification of soluble protein of seashell nacre is carried out by: washing, acid treating, crushing, dialyzing, centrifuging, and drying. Furthermore, It is desalted with FPLC and is separated and purified by anionic exchange chromography. Within each cycle of 20min, 0.3-0.6mg purified soluble shell protein is obtained. It is used to induce human bone generation and to promote bone renovation in clinic. Meanwhile, it provides a technology of in vitro growth of valuable pearl.

Description

technical field [0001] The invention relates to a method for separating and purifying a large amount of shell protein in a short time by using a fast protein chromatography system (FPLC). The produced product can be widely used in the research and production of biological materials and in vitro cultured pearls. Background technique [0002] Discussing the formation mechanism of shells opens a new way of thinking for the development of new materials such as nanomaterials, mineral polymer materials and template crystals. Soluble shell protein (SMP) plays a decisive role in shell formation, regulating CaCO 3 Crystal deposition, including nucleation, growth, phase transition and crystal morphology of calcite and aragonite crystals. Studies in recent years have shown that shell protein can promote the differentiation of human osteoblasts, induce bone formation, and promote bone repair, so it is gradually applied to materials medicine. The extraction of soluble shell protein is ...

Claims

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Application Information

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IPC IPC(8): C07K1/18C07K14/435
Inventor 张文兵麦康森徐玮李静董瑞娜赵建民谭北平艾庆辉刘付志国马洪明
Owner OCEAN UNIV OF CHINA
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