THE IMPACT OF THE AMOUNT OF HISTIDINE IN C-TERMINAL FUSION-TAG ON THE SECRETION OF ENDOGLUCANASE B IN BACILLUS SUBTILIS
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Abstract
In this research, the host cell Bacillus subtilis and the reporter gene celB were used for evaluating the impact of the amount of histidine in C-terminal fusion tag on the secretion of Endogucanase B. The results show that B. subtilis 1012 is more appropriate for secreting CelB than B. subtilis WB800N. Among three kinds of His tag (4His, 5His, 6His), 6His tag has the most impacts on decreasing the secretion of CelB. In contrast, the surveying of binding capacity of these proteins demonstrates that CelB-6His has the best binding capacity and the fewest leakage (4.78%) while CelB-4His has the most leakage (16.1%) in purifying process by Ni-NTA.
Keywords
Bacillus subtilis, Histidine, Endoglucanse B, Pgrac212, fusion tag