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毛果杨中链酰基辅酶A合成酶的克隆及酶学分析

曹山 蒋璐瑶 李丽红 要笑云 张强 撖静宜 王莹 李慧 陆海

曹山, 蒋璐瑶, 李丽红, 要笑云, 张强, 撖静宜, 王莹, 李慧, 陆海. 毛果杨中链酰基辅酶A合成酶的克隆及酶学分析[J]. 北京林业大学学报, 2016, 38(7): 9-15. doi: 10.13332/j.1000-1522.20160121
引用本文: 曹山, 蒋璐瑶, 李丽红, 要笑云, 张强, 撖静宜, 王莹, 李慧, 陆海. 毛果杨中链酰基辅酶A合成酶的克隆及酶学分析[J]. 北京林业大学学报, 2016, 38(7): 9-15. doi: 10.13332/j.1000-1522.20160121
CAO Shan, JIANG Lu-yao, LI Li-hong, YAO Xiao-yun, ZHANG Qiang, HAN Jing-yi, WANG Ying, LI Hui, LU Hai.. Cloning and enzymatic analysis of medium-chain acyl coenzyme A synthetase in Populus trichocarpa.[J]. Journal of Beijing Forestry University, 2016, 38(7): 9-15. doi: 10.13332/j.1000-1522.20160121
Citation: CAO Shan, JIANG Lu-yao, LI Li-hong, YAO Xiao-yun, ZHANG Qiang, HAN Jing-yi, WANG Ying, LI Hui, LU Hai.. Cloning and enzymatic analysis of medium-chain acyl coenzyme A synthetase in Populus trichocarpa.[J]. Journal of Beijing Forestry University, 2016, 38(7): 9-15. doi: 10.13332/j.1000-1522.20160121

毛果杨中链酰基辅酶A合成酶的克隆及酶学分析

doi: 10.13332/j.1000-1522.20160121
基金项目: 

国家自然科学基金项目(31570582、30671697、J1103516)。

详细信息
    作者简介:

    曹山,博士生。主要研究方向:树木分子生物学。Email:llcs0909@163.com地址:100083北京市海淀区清华东路35号北京林业大学生物科学与技术学院。责任作者:陆海,教授,博士生导师。主要研究方向:树木发育与木材形成。Email:luhai1974@bjfu.edu.cn地址:同上。

    曹山,博士生。主要研究方向:树木分子生物学。Email:llcs0909@163.com地址:100083北京市海淀区清华东路35号北京林业大学生物科学与技术学院。责任作者:陆海,教授,博士生导师。主要研究方向:树木发育与木材形成。Email:luhai1974@bjfu.edu.cn地址:同上。

Cloning and enzymatic analysis of medium-chain acyl coenzyme A synthetase in Populus trichocarpa.

  • 摘要: 中链酰基辅酶A合成酶(MACS)属于腺苷酸合成酶超基因家族,催化中链脂肪酸与辅酶A结合形成相应的中链酰基辅酶A。本研究通过同源检索毛果杨基因组数据库中的4CL基因,克隆得到毛果杨PtMACS1的基因序列(基因编号:estExt_fgenesh4_pg.c_640066)。通过序列分析可知,Box I和Box II两个在4CL中保守的结构域在该蛋白中并不保守。将PtMACS1与原核表达载体 pET-30a(+) 构建融合表达载体PtMACS1- pET-30a(+),转化大肠杆菌BL21(DE3)后诱导重组蛋白进行表达。镍柱纯化重组蛋白后进行酶学活性分析,研究结果表明该蛋白对中链脂肪酸己酸、壬酸、癸酸显示出明显活性:Kcat分别为(1302.65)、(1933.46)、(2015.51) mol/(minmg),以该蛋白的最适底物癸酸测得该蛋白的最适反应温度为37 ℃,最适反应pH为7.0。该结果表明,PtMACS1属于毛果杨中链酰基辅酶A合成酶家族一员,为后续毛果杨腺苷酸合成酶超基因家族的鉴定及分类分析提供资料。

     

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出版历程
  • 收稿日期:  2016-04-08
  • 刊出日期:  2016-07-30

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