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Extraction and purification of matrix protein from the nacre of pearl oyster pinetada fueata

机译:珍珠贝珍珠母珍珠质中基质蛋白的提取与纯化

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A soluble matrix protein P14 with an apparent molecular mass of 14.5 kDa was isolated from fragmented nacre of pearl oysters (Pinctada fucata) treated with 10% NaOH solution to investigate the nacre matrix proteins and their effect on the CaC03 crystal. The protein was characterized by gel exclusion chromatography and reversed-phase high performance liquid chromatography after demineralization by 10% acetic acid. The X-ray diffraction pattern of P14 crystals indicates that P14 plays an important role in nacre biomineralization. P14 can induce aragonite formation, stimulate CaC03 crystal formation, and accelerate aragonite precipitation. Heating of the acid insoluble nacre residue, which was named conchiolin, in 10% sodium dodecyl sulfate solution supplemented with 10% β-mercaptoethanol solution for 10–20 min at about 100°C gave two other soluble proteins having molecular masses of 19.4 kDa and 25.0 kDa. The present study suggests that these two proteins are linked to the insoluble organic matrix by disulfide bridges because the extraction yield increases when β-mercaptoethanol is added to the medium.
机译:从表观分子质量为14.5 kDa的可溶性基质蛋白P14中分离出10%NaOH溶液处理的珍珠牡蛎珍珠碎片,以研究珍珠母基质蛋白及其对CaC0 3 水晶。 10%乙酸脱盐后,通过凝胶排阻色谱和反相高效液相色谱对蛋白质进行表征。 P14晶体的X射线衍射图表明P14在珍珠质生物矿化中起重要作用。 P14可以诱导文石的形成,刺激CaCO3 3 晶体的形成,并加速文石的沉淀。在大约100°C的温度下,在10%十二烷基硫酸钠溶液中添加10%β-巯基乙醇溶液,加热酸不溶性珍珠母残渣(称为conchiolin),加热10-20分钟,得到另外两种分子量为19.4 kDa的可溶性蛋白质, 25.0 kDa。本研究表明,这两种蛋白质通过二硫键连接至不溶性有机基质,因为当将β-巯基乙醇添加到培养基中时,提取产率会提高。

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