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    • 4. 发明授权
    • One-step purification process for organophosphorus hydrolase enzyme
    • 有机磷水解酶的一步纯化方法
    • US06469145B1
    • 2002-10-22
    • US09603450
    • 2000-06-26
    • Vipin K. RastogiTu-Chen ChengJoseph J. DeFrank
    • Vipin K. RastogiTu-Chen ChengJoseph J. DeFrank
    • C08H100
    • C12N9/16
    • Novel and improved processes for isolating organophosphorus hydrolase enzyme from an aqueous solution and obtaining substantially purified enzyme at high yield are provided, as well as compositions, including storage stable lyophilyzed organophosphorus hydrolase enzyme compositions, that are prepared by the provided methods. The organophosphorus hydrolase enzyme is purified by contacting an aqueous solution of cell free bacterial proteins with a strong cation exchange resin, the aqueous solution comprising soluble organophosphorus hydrolase enzyme, washing the strong cation exchange resin with a washing buffer to remove unbound proteins from the strong cation exchange resin, eluting proteins that remain bound to the strong cation exchange resin by washing the resin with an eluting buffer comprising salt in a concentration that starts at about zero and is raised during the eluting process to about 0.5M, and detecting and collecting eluate comprising a protein having organophosphorus hydrolase enzyme activity. All processing steps are conducted at a pH that is less than the isoelectric point of the organophosphorus hydrolase enzyme.
    • 提供了从水溶液中分离有机磷水解酶并以高产率获得基本上纯化的酶的新颖和改进的方法,以及通过所提供的方法制备的组合物,包括储存稳定的冻干有机磷水解酶组合物。 通过使无细胞细菌蛋白质的水溶液与强阳离子交换树脂接触来纯化有机磷酸水解酶,该水溶液包含可溶性有机磷水解酶,用洗涤缓冲液洗涤强阳离子交换树脂以从强阳离子去除未结合的蛋白质 通过用洗脱缓冲液洗涤树脂,所述洗脱缓冲液含有浓度从约零开始并在洗脱过程中升高至约0.5M的盐,并检测和收集洗脱液,所述洗脱缓冲液包含 具有有机磷水解酶活性的蛋白质。 所有处理步骤在小于有机磷水解酶的等电点的pH下进行。