嗜热菌Geobacillus Kaustophilus HTA426磷酸三酯酶在毕赤酵母GS115中的表达  

Overexpression of Phosphotriesterase( PTE) Gene from thermophilic Geobacillus Kaustophilus HTA426 in Pichia pastoris GS115

作　　者：詹冬玲[1] 林禹欣[1] 任玉雪[1] 刘洋[2] 

机构地区：[1]吉林农业大学食品科学与工程学院,吉林长春130118 [2]吉林化工学院化学与制药工程学院,吉林吉林132022

出　　处：《吉林化工学院学报》2013年第11期17-21,共5页Journal of Jilin Institute of Chemical Technology

基　　金：吉林农业大学博士启动基金项目(201223)

摘　　要：将嗜热菌Geobacillus Kaustophilus HTA426中的磷酸三酯酶基因转入毕赤酵母GS115中,整合到染色体DNA基因组上,并进行诱导表达.SDS-PAGE和Western-blot试验结果显示,重组蛋白是一个分子量约为50KD的磷酸三酯酶二聚体.酶活检测证明重组蛋白具有较高的磷酸三酯酶活性,其最适温度为70℃,最适pH为10.0.The ground-state geometries of 8-Hydroxyquinoline Lithium and its derivatives were optimized using DNF/B3LYP method with 6-31g ＊ basis set. On the basis of the ground states, the absorption spectra were obtained by time-dependent DFY （TD-DFT）. The lowest lying singlet excited state of 8-Hydroxyquinoline Lithium was fully optimized at the C[S/6-3 lg ＊ levels. The results indicate that introducing one amino group on the 2 point of the quinoline group and introducing amino and hydroxy groups on the 2 and 5 point of the quinoline group make the dihedral angle increase, it suggest that the electron donors can decrease the conjugated degree. The introduction of the electron-withdrawing group lows down the HOMO and LUMO energy levels and their energy gaps of the complexes, so their absorption spectra are red shift compared with 8- Hydroxyquinoline Lithium. The large stocks shift of 108. 43 nm of 8-Hydroxyquinoline Lithium may be explained by its large change of the structures between the ground and the excited state.

关 键 词：嗜热菌 磷酸三酯酶 毕赤酵母GS115 表达 

分 类 号：Q786[生物学—分子生物学]