质粒平均分配基因parDE对甲酸脱氢酶NADH再生系统稳定性的影响

›› 2008, Vol. 8 ›› Issue (2): 345-349.

质粒平均分配基因parDE对甲酸脱氢酶NADH再生系统稳定性的影响

张刚杨光裴海生李春曹竹安

清华大学化学工程系生物化工研究所清华大学化学工程系生物化工研究所清华大学生物化工研究所北京理工大学生命科学与技术学院生物技术系清华大学化工系生物化工研究所

收稿日期:2007-10-22 修回日期:2008-01-07 出版日期:2008-04-20 发布日期:2008-04-20
通讯作者: 曹竹安

Effect of Plasmid Distribution Gene parDE on the Stability of Formate Dehydrogenase NADH Regeneration System

ZHANG Gang YANG Guang LI Chun CAO Zhu-an

Department of Biotechnology, School of Life Science and Technology, Beijing Institute of Technology Inst. Biochem. Eng., Dept. Chem. Eng., Tsinghua University

Received:2007-10-22 Revised:2008-01-07 Online:2008-04-20 Published:2008-04-20
Contact: CAO Zhu-an

摘要/Abstract

摘要： 将质粒平均分配基因parDE引入重组质粒pDK7-fdh(携带甲酸脱氢酶基因fdh)中，得到基因重组菌F6. 考察了F6与未引入parDE基因的基因重组菌FY(含重组质粒pDK6-fdh)和F-1(含重组质粒pMAL-p2X-fdh)的质粒遗传稳定性及甲酸脱氢酶活性的差异. 结果显示，F6传代160代后，仅有3%的菌质粒丢失，而基因重组菌F-1和FY(均只携带fdh基因)传代160代后，丢失质粒的菌分别占93%和78%. 传代160代后，F6的甲酸脱氢酶比酶活为2.77 U/mg，比传代前降低1.77%, FY为2.02 U/mg, F-1与野生型菌株M5al相近，为1.33 U/mg. 结果表明，引入parDE基因的基因重组菌质粒稳定性和甲酸脱氢酶活性均远高于未引入parDE基因的基因重组菌.

关键词: 质粒稳定性, parDE基因, fdh基因, 比酶活

Abstract: The recombinant bacterium F6 was constructed with the recombinant plasmid pDK7-fdh-parDE which contained the genes parDE and fdh. While the FY and F-1 were constructed with the recombinant plasmids pDK6-fdh and pMAL-p2X-fdh only containing fdh, respectively. K. oxytoca M5al is the initial bacterium of F6, FY and F-1. The effect of parDE on plasmid stability and FDH (formate dehydrogenase) enzyme activity was also studied. It was shown that 93% F-1 and 78% FY lost the plasmids while only 3% F6 lost the plasmids after 160 generations. Furthermore, the FDH specific enzyme activity of F6 decreased from 2.82 to 2.77 U/mg, only 1.77% decreased, while the enzyme activity of FY decreased significantly from 2.69 to 2.02 U/mg, and F-1 decreased from 2.63 to 1.33 U/mg, close to the enzyme activity of M5al after 160 generations. It was indicated that the recombinant bacteria with the parDE had obvious advantage on plasmid stability, and FDH enzyme activity was higher than the recombinant bacteria without the parDE.

Key words: plasmid stability, parDE gene, fdh gene, specific enzyme activity

中图分类号:

Q789
TQ923

张刚杨光裴海生李春曹竹安. 质粒平均分配基因parDE对甲酸脱氢酶NADH再生系统稳定性的影响[J]. , 2008, 8(2): 345-349.

ZHANG Gang YANG Guang LI Chun CAO Zhu-an. Effect of Plasmid Distribution Gene parDE on the Stability of Formate Dehydrogenase NADH Regeneration System[J]. , 2008, 8(2): 345-349.