猪肌肉葡萄糖6-磷酸异构酶的分离纯化及性质研究

CORC > 集美大学

	猪肌肉葡萄糖6-磷酸异构酶的分离纯化及性质研究
	周剑 ; 孙乐常 ; 许慧中 ; 汪宁 ; 张凌晶 ; 曹敏杰
刊名	http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=spkj200910042&dbcode=CJFQ&dbname=CJFQ2009
	2012-06-05 ; 2012-06-05
关键词	猪葡萄糖6-磷酸异构酶分离纯化性质 pig glucose-6-phosphate isomerase purification characterization Q814
其他题名	Purification and characterization of glucose-6-phosphate isomerase from pig skeletal muscle
中文摘要	葡萄糖6-磷酸异构酶(Glucose-6-phosphate isom erase,GPI;EC5.3.1.9)能催化果糖6-磷酸(fructose-6-phosphate,F-6-P)异构为葡萄糖6-磷酸(glucose-6-phosphate,G-6-P),是糖原酵解过程中的重要酶之一。本研究通过硫酸铵盐析、DEAE-Sepharose、SP-Sepharose以及Hydroxyapatite层析等方法从猪肌肉中分离纯化得到GPI,纯化倍数为105.6,比活力为159.4U/mg。SDS-PAGE和凝胶过滤层析表明猪GPI为单体蛋白,分子量约为55kDa。免疫印迹检测结果显示,猪GPI能够和抗白姑鱼GPI多克隆抗体发生免疫交叉反应。酶学性质显示猪GPI的最适温度为50℃,最适pH为8.5。在pH8.5、25℃条件下测得其Km值为0.39mmol/L,最大反应速率Vmax为0.61mmol/L·min。; Glucose-6-phosphate isomerase(GPI;EC 5.3.1.9) is a key enzyme of glycogen metabolism. It catalyses the second step of glycolysis,the interconversion of glucose-6-phosphate(G-6-P) and fructose-6-phosphate(F-6-P). In the present study,GPI was purified to homogeneity from pig skeletal muscle by ammonium sulfate fractionation,column chromatographies on DEAE-Sepharose,SP-Sepharose and Hydroxyapatite. GPI was purified 105.6-fold and its specific activity was 159.4U/mg. The purified enzyme was a monomer with a molecular mass of 55kDa as determined by SDS-PAGE and gel filtration. Western blot analysis revealed that pig GPI cross reacted with anti-white croaker GPI polyclonal antibody. The optimum pH and temperature of pig GPI was 8.5 and 50℃,respectively for the isomerization of F-6-P to G-6-P. The Michaelis-Menten constant(Km) was 0.39mmol/L and the maximum velocity(Vmax) was 0.61mmol/L·min at pH 8.5 and 25℃.; 【作者单位】福建省高校水产科学技术与食品安全重点实验室,集美大学生物工程学院；上海海洋大学食品学院；【作者英文名】ZHOU Jian1,2,SUN Le-chang1,XU Hui-zhong1,WANG Ning1,2,ZHANG Lin-jing1,CAO Min-jie1,(1. The Key Laboratory of Science and Technology for Aquaculture and Food Safety,School of Biotechnology,Jimei University,Xiamen 361021,China;2. College of Food Scienc
语种	中文
内容类型	期刊论文
源URL	[http://ir.calis.edu.cn/hdl/235041/18303]
专题	集美大学
推荐引用方式 GB/T 7714	周剑,孙乐常,许慧中,等. 猪肌肉葡萄糖6-磷酸异构酶的分离纯化及性质研究[J]. http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=spkj200910042&dbcode=CJFQ&dbname=CJFQ2009,2012, 2012.
APA	周剑,孙乐常,许慧中,汪宁,张凌晶,&曹敏杰.(2012).猪肌肉葡萄糖6-磷酸异构酶的分离纯化及性质研究.http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=spkj200910042&dbcode=CJFQ&dbname=CJFQ2009.
MLA	周剑,et al."猪肌肉葡萄糖6-磷酸异构酶的分离纯化及性质研究".http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=spkj200910042&dbcode=CJFQ&dbname=CJFQ2009 (2012).