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鸡胸肉中钙激活酶的纯化研究

DOI: 10.7685/j.issn.1000-2030.2013.06.017, PP. 105-110

Keywords: 鸡胸肉,钙激活酶,阴离子交换,疏水性层析,纯化

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Abstract:

对宰后10min的鸡胸肉经匀浆后利用超高速冷冻离心、硫酸铵分级沉淀、离子交换层析和疏水层析等技术,从中分离纯化出μ-钙激活酶(μ-calpain)和μ/m-钙激活酶(μ/m-calpain)。对分离纯化过程各主要步骤中的酶总活性、比活性等参数进行测定,同时采用活性电泳对纯化过程的样品进行分析。结果表明样品通过HitrapCaptoDEAE层析柱,可以将μ-calpain和μ/m-calpain分别洗脱下来,μ-calpain比活性为1.02U?mg-1,μ/m-calpain的比活性为3.04U?mg-1;所得μ-calpain及μ/m-calpain样品分别依次经过PhenylSepharose6FF层析柱和MonoQ层析柱,最后得到的μ-calpain和μ/m-calpain比活性分别为26.89和34.96U?mg-1。结果表明,随着纯化步骤的增加,鸡胸肉中的钙激活酶纯度逐步增加,并得到了较有效的纯化。

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