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首页> 美国政府科技报告 >1000-L Scale-Up Fermentation of Escherichia Coli Containing PVSEOP7 for Production of Organophosphorus Hydrolase
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1000-L Scale-Up Fermentation of Escherichia Coli Containing PVSEOP7 for Production of Organophosphorus Hydrolase

机译：含有pVsEOp7的大肠杆菌1000-L放大发酵生产有机磷水解酶

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Four 20-L scale batch fermentations in LB medium were performed to investigate time-dependent yields of organophosphorus hydrolase (OPH) in E. coli XL1 containing pVSEOP7. The specific OPH activity was monitored during each 20-L batch fermentation culture by assaying several times between the IPTG induction point (OD of 0.4 to 0.5 at 600 nm) and the cell harvest point (near- stationary phase of each 20-L batch culture). From the four 20-L batch fermentations, only the 20-L batch cultures with 1 mM of cobalt chloride added 3-hr prior to cell harvest resulted in a drastic increase in the specific OPH activity (369 to 1809 U/mL). A similar strategy of 1 mM cobalt chloride addition was successfully used to obtain a 20.7 mg OPH per liter of culture at the 1000-L scale batch fermentation in LB, as compared to 5 to 10 mg/L in flask cultures. The specific OPH activity was shown to be stable through additional process time associated with downstream processing steps such as cell concentration by continuous disk centrifugation, cell paste preparation by bottle centrifugation, cell disruption by Microfluidizer homogenization, and lysed cell clarification by 0.22 m tangential flow microfiltration.

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