摘要
脂氧合酶(LOX)是植物十八碳酸途径中一个很重要的酶,该酶作用的产物在植物的生长发育过程中以及在植物对环境胁迫反应中起着重要的作用.通过PCR和酶切连接的方法,构建了乳突果脂氧合酶基因的原核表达载体,将其导入大肠杆菌BL21(DE3)中,在低温(15℃)条件下,表达出了有活性的酶蛋白,且该蛋白分子量与预期分子量相符.通过LC-ESI-MS检测,该酶可催化亚油酸生成9,10,11-三羟基-12-十八碳烯酸.推测在乳突果中,几丁质诱导的十八碳酸代谢途径是沿着9-LOX方向进行的.
Lipoxygenases are important enzymes in plant octadecanoic acid pathway.Products of this enzyme have important functions in plant growth and development,as well as in response to environmental stress.A prokaryotic expression vector of lipoxygenase gene from Adelostemma gracillimum was constructed through PCR and enzyme digestion connection method and a full length lipoxygenase cDNA was successfully expressed in E.coli BL21（DE3）under low temperature（15 ℃）.Molecular weight of the recombinant protein was the same as the expected.Results of enzyme assay showed that the protein could catalyze linoleic acid to 9,10,11-trihydroxy-12-octadecenoic acid detected by LC-ESI-MS.It was speculated that octadecanoic acid pathway induced by chitosan differentiated to 9-LOX orientation.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2012年第5期156-160,共5页
Journal of Gansu Agricultural University
基金
云南省自然科学基金项目(2009ZC085M)
关键词
乳突果
脂氧合酶
十八碳酸途径
原核表达
酶活测定
Adelostemma gracillimum
lipoxygenase
octadecanoic acid pathway
prokaryotic expression
enzyme activity assay
