| 猪肺血管紧张素转化酶的提取与纯化 |
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| 引用本文： | 周利琴,廖丹葵,孙建华,伍善广,兰雄雕,周敬豪,徐存华.猪肺血管紧张素转化酶的提取与纯化[J].广西农业科学,2014(4):639-642. |
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| 作者姓名： | 周利琴 廖丹葵 孙建华 伍善广 兰雄雕 周敬豪 徐存华 |
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| 作者单位： | 广西大学化学化工学院,南宁530004 |
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| 基金项目： | 广西自然科学基金项目（2012GXNSFDA053004）;广西生物炼制重点实验室开放基金项目（GXBF11-06）;广西研究生创新计划项目（YCBZ2012010） |
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| 摘 要： | 目的]研究猪肺血管紧张素转化酶（ACE）的提纯方法,为其生产利用提供技术支撑.方法]以新鲜猪肺组织为试验材料,匀浆离心后经硫酸铵分级沉淀透析,然后采用离子交换-超滤离心联合法对猪肺ACE粗提物进行纯化,测定ACE的分子量、酶活力、纯化倍数及酶活力回收率.结果]通过离子交换-超滤离心联合法分离纯化猪肺ACE,可得到相对分子量182 kDa、酶比活力1.2476 U/mg的电泳级纯品ACE,其纯化倍数达357.31倍,酶活力回收率达12.28％.ACE催化反应动力学米氏常数（Km）为0.849 mmol/L,最大反应速率（Vmax）为9.775 nmol/min.结论]采用离子交换-超滤离心联合法纯化猪肺ACE能够极大缩短分离与纯化时间,且获得较高的纯化倍数和酶活力回收率,是猪肺ACE的高效纯化方法.
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| 关 键 词： | 血管紧张素转化酶 猪肺 阴离子交换 超滤离心 纯化 |
Extraction and purification of angiotensin converting enzyme from fresh hog lung |
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| ZHOU Li-qin,LIAO Dan-kui,SUN Jian-hua,WU Shan-guang,LAN Xiong-diao,ZHOU Jing-hao,XU Cun-hua.Extraction and purification of angiotensin converting enzyme from fresh hog lung[J].Guangxi Agricultural Sciences,2014(4):639-642. |
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| Authors: | ZHOU Li-qin LIAO Dan-kui SUN Jian-hua WU Shan-guang LAN Xiong-diao ZHOU Jing-hao XU Cun-hua |
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| Institution: | (School of Chemistry and Chemical Engineering, Guangxi University, Nanning 530004, China) |
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| Abstract: | Objective]Extraction and purification of angiotensin converting enzyme （ACE） was studied to provide techni- cal support for its production and utilization. Method]ACE crude extracts were purified by DEAE-Sepharose FF anion ex- change and ultrafihration after the fresh lung homogenate was centrifuged, precipitated and dialyzed using ammonium sulfate. ACE molecular weight, specific activity, purification fold and activity recovery were determined. Result]Lung ACE obtained by SDS-PAGE showed ACE-pure and molecular mass was 182kDa. The ACE was purified to 357.31-fold with specific activity of 1.2476 U/mg and activity recovery of 12.28%. The Michaelis-constant of the ACE was 0.849 mmol]L and Vmax was 9.775 nmol/min. Conclusion]The ion exchange-uhrafiltration is an efficient purification method for lung ACE, as it can greatly shorten the separation and purification time, and get a higher purification fold and activity recovery. |
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| Keywords: | angiotensin converting enzyme hog lung anion exchange ultrafiltration centrifugation purification |
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