Shuxia Shen
College of Food Science and Engineering，Jilin Agricultural University，Changchun 130118，Jilin Province，ChinaYunming Zhu
College of Food Science and Engineering，Jilin Agricultural University，Changchun 130118，Jilin Province，ChinaWeihong Min
College of Food Science and Engineering，Jilin Agricultural University，Changchun 130118，Jilin Province，ChinaLi Fang
College of Food Science and Engineering，Jilin Agricultural University，Changchun 130118，Jilin Province，ChinaJinkun Xu
College of Food Science and Engineering，Jilin Agricultural University，Changchun 130118，Jilin Province，ChinaSupported by the Natural Science Foundation of Jilin Province ( 20130101139JC) and by the National Science and Technology Support Program ( 2012BAD37B05)
Abstract:［Objective］To obtain a new homoserine dehydrogenase with better properties from Corynebacterium pekinense by the spatial structure transfromation．［Methods］Double mutants L200F/D215A，L200F/D215E，L200F/D215G and L200F/D215K were constructed by site-directed mutagenesis and expressed in E．coli BL21．L200F /D215K was characterized for its highest catalytic efficiency and compared with that of L200F．［Results］The Vmax of L200F/D215K was 36.92 U/mg，1.24 times as that of L200F．The optimum reaction temperature of L200F/D215K was 37℃，2℃ higher than that of L200F．The optimum pH of L200F/D215K was 7.5，the same as that of L200F．The half-life time of L200F/D215K under optimum temperature was 4.16 h and was 1.12 times as that of L200F．Both L200F/D215K and L200F had good resistance to organic solvents and metal ions．［Conclusion］Through the spatial structure transformation，the enzymatic activity was increased，and the enzymology properties was optimized．
Shuxia Shen, Yunming Zhu, Weihong Min, Li Fang, Jinkun Xu. Heterologous expression and characterization of L200F/D215K mutant of homoserine dehydrogenase from Corynebacterium pekinense AS1.299. [J]. Acta Microbiologica Sinica, 2014, 54(10): 1178-1184
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