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Position: Home > Articles > The purification of calpains from chicken breast meat Journal of Nanjing Agricultural University 2013,36 (6) 105-110

鸡胸肉中钙激活酶的纯化研究

作  者:
慕妮;王恺;王璐莎;黄明;巴吐尔·阿不力克木
单  位:
新疆农业大学食品科学与药学学院;南京农业大学肉品加工与质量控制教育部重点实验室
关键词:
鸡胸肉;钙激活酶;阴离子交换;疏水性层析;纯化
摘  要:
对宰后10 min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Hitrap Capto DEAE层析柱,可以将μ-calpain和μ/m-calpain分别洗脱下来,μ-calpain比活性为1.02 U·mg-1,μ/m-calpain的比活性为3.04 U·mg-1;所得μ-calpain及μ/m-calpain样品分别依次经过Phenyl Sepharose 6 FF层析柱和Mono Q层析柱,最后得到的μ-calpain和μ/m-calpain比活性分别为26.89和34.96 U·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。
译  名:
The purification of calpains from chicken breast meat
作  者:
MU Ni;WANG Kai;WANG Lusha;HUANG Ming;BATUER Abulikemu;Key Laboratory of Meat Processing and Quality Control,Ministry of Education,Nanjing Agricultural University;College of Food Science and Pharmaceutical Science, Xinjiang Agricultural University;
关键词:
chicken breast meat;;calpain;;anion-exchange chromatography;;hydrophobic chromatography;;purification
摘  要:
Chicken breast meat( within 10 min postmortem) was sampled and treated with procedures such as homogenization,ammonium sulfate fractionation,ionic exchange,hydrophobic chromatography,etc. The total activity and specific activity of calpains and other parameters related to different major purification procedures were detected. Meanwhile,the samples in purification processes were verified by way of casein zymography. It showed that μ-calpain and μ / m-calpain in the sample could be separated through Hitrap Capto DEAE anion-exchange column. In this step,the specific activity of μ-calpain was 1. 02 U·mg- 1and that of μ / m-calpain was 3. 04U·mg- 1. Then,the sample containing μ-calpain went through Phenyl Sepharose 6 FF Hydrophobic column and Mono Q anion-exchange column one by one,and so did μ / m-calpain sample. In the last step,the specific activity of resultant μ-calpain was 26. 89 U·mg- 1and that of μ / m-calpain was 34. 96 U·mg- 1. It confirmed that as the purification step increased,the purity of calpains from chicken breast meat was raised gradually and calpains were purified effectively in this purification processes.

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