重组有机磷水解酶MPH的发酵条件的优化研究

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。

Optimization of fermentation condition for remobilization organophosphate hydrolase MPH

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 fermentation 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.