   2.4.1.162 | recombinant C-terminally Strep-tagged enzyme from Escherichia coli strain BL21(DE3) Rosetta inclusion bodies, method optimization and evaluation of a cost effective renaturation, detailed overview. The enzyme is solubilized from batch sample by detergent buffer containing 1 M urea, 0.1 M Tris, 25 mM deoxycholate, and 1% v/v IGEPAL CA-630. Refolding is done by batchwise or continuous exchange of dialysis buffer with cellulose membrane, over a period of 48 h and a buffer to sample volume ratio of 100:1 at 4°C, batch dialysis. Mild solubilization at low urea concentration (2 M) and high pH (pH 12) does not improve the recovery of protein from inclusion bodies. Phosphate buffers exhibit above-average activity yield of 1022 U/ml |
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