【摘要】 通过对毛白杨中的4-香豆酸:辅酶A连接酶1(Pt4CL1)蛋白第338位缬氨酸进行缺失突变,从而获得了具有芥子酸催化活性的Pt4CL1蛋白突变体338dVal。与野生型4CL1蛋白活性相比较,该突变体获得了催化芥子酸的活性,比活力是(1.62±0.34)nkat/mg。同时,对4-香豆酸催化活性有轻微的降低(约降低12%),对咖啡酸的催化活性有明显增加(约增加126%),对阿魏酸的催化活性有部分的降低(约降低29%),对肉桂酸的催化活性没有显著变化。该结论证实了第338位缬氨酸对底物芥子酸5位甲氧基具有空间位阻效应。该项研究为通过基因工程技术调控木质素的合成提供了新的方法。更多还原

【Abstract】 4-coumarate:CoA ligase 1(4CL1) is a key rate-limiting enzyme in the pathway of lignin biosynthesis.Studying the catalytic properties of 4CL1 enzyme is useful for understanding the pathway of lignin biosynthesis.This paper reports that a Pt4CL1 variant 338dVal,which is a deleted Val-338 in amino acid sequence of 4-coumarate:CoA ligase from Populus tomentosa,gains the function of using sinapic acid.The specific activity to sinapic acid was(1.62±0.34) nkat/mg.Meanwhile,the catalytic activity of 338dVal to 4-coumarate acid decreased by 12% approximately;the catalytic activity of 338dVal to caffeic acid increased by 126% and the catalytic activity of 338dVal to ferulic acid decreased by 29% roughly.The catalytic acitivity of 338dVal to cinnamic acid did not change.The results show that the Val-338 in Pt4CL1 has size exclusion effect on 5-methoxy of sinapic acid.The paper provides a new method that can be used to regulate the lignin biosynthesis using genetic engineering.更多还原