摘要
有机磷水解酶在去除有机磷农药残留中具有重要的应用前景。在Escherichia coli BL21(DE3)中实现了有机磷水解酶(MPH)的重组诱导表达之后,为了实现工业化生产MPH,考察了以乳糖为诱导剂,碳源、氮源、金属离子、培养温度、乳糖浓度及诱导时间等对产酶的影响,得到了优化的发酵条件,L9(34)正交实验进一步确定了最佳的碳源、氮源及乳糖浓度。在此基础上利用7L自控发酵罐进行了发酵过程研究,经12h培养,得到菌体12.65g(DCW)/L,MPH表达量为14.56%,酶活18.69I U/mL。
Organophosphate hydrolase has great potential in removal of organophosphate compounds. An organophosphate hydrolase MPH was expressed in Escherichia coli BL21 （DE3） . Experiments were designed to optimize the parameters for fermemation of MPH. Inexpensive lactose was used as an inducer of expression of the MPD gene, which is under the control of a trc promoter. Composition of the culture medium, lactose concentration, induction phase and metal ions were tested, L9 orthogonal test was designed to determine the optimal carbon and nitrogen source and lactose concentration. Fed-batch culture process of E. coli BL21 （DE3）/pET29a-mpd were carried out to produce recombinant MPH in 7L Bioengineering autocontrol fermentor . The final cell density and concentration of MPH fusion were 12.65g（DCW）/L and 14.56% respectively.
出处
《工业微生物》
CAS
CSCD
北大核心
2007年第5期27-32,共6页
Industrial Microbiology
基金
国家863计划(2004AA246070)
国家自然科学基金(30300005)资助项目
关键词
甲基对硫磷水解酶
乳糖
正交实验
条件优化
methyl parathion hydrolase
lactose
orthogonal test
optimization of cultural condition
