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Updated: Mar 26, 2026

Overexpressing and Purifying a Toxic Nuclease from Escherichia coli
Published on: August 29, 2025
Selection of suitable detergents for obtaining an active dengue protease in its natural form from E. coli
Lynette Sin Yee Liew1, Michelle Yueqi Lee1, Ying Lei Wong1
1Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31Biopolis Way, Nanos, #03-01, 138669, Singapore.
Abstract:
Dengue protease is a two-component enzyme and is an important drug target against dengue virus. The protease activity and protein stability of dengue nonstructural protein 3 (NS3) require a co-factor region from a four-span membrane protein NS2B. A natural form of dengue protease containing full-length NS2B and NS3 protease domain NS2BFL-NS3pro will be useful for dengue drug discovery. In current study, detergents that can be used for protease purification were tested. Using a water soluble protease construct, 39 detergents were selected for both NS2B and NS2BFL-NS3pro purification. The results showed that 18 detergents were able to sustain the activity of the natural dengue protease and 11 detergents could be used for NS2B purification. The results obtained in this study will be useful for biochemical and biophysical studies on dengue protease.
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