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Author:
Date:
2014-11-05
[Abstract] The purification of a target protein from a complex mixture of proteins is a challenging undertaking. If the target protein has been previously characterised, then information such as subcellular location, function, molecular weight and pI can be used for the design of a purification strategy. However, if the target protein is uncharacterised or little information regarding its characteristics is available, a generic purification protocol can be employed that is optimised as additional characteristics of the target protein are determined during subsequent purification steps. Herein, we describe the protocol for the purification and detection of a poly-γ-D-glutamic acid (PDGA) depolymerase from a consortium culture of two Gram-negative bacteria using a combination of chromatography, ...
[摘要] 从蛋白质的复杂混合物中纯化靶蛋白是一项具有挑战性的任务。 如果靶蛋白以前已被表征，则诸如亚细胞定位，功能，分子量和pI的信息可用于设计纯化策略。 然而，如果目标蛋白质是未表征的或者关于其特征的很少的信息可用，则可以使用通用的纯化方案，其随着在随后的纯化步骤期间测定靶标蛋白质的附加特征而被优化。 在这里，我们描述了从两个革兰氏阴性细菌的联合培养物使用色谱，2D电泳和酶谱的组合纯化和检测聚-γ-D-谷氨酸（PDGA）解聚酶的协议。
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