Abstract
Subtilisin is an alkaline serine protease produced by a wide variety of Bacillus species. In view of the industrial application for detergent and food processing, subtilisin has been the subjest of extensive protein engineering efforts.1 We have previously cloned and expressed the gene of subtilisin E from Bacillus subtilis into Escherichia coli.2,3 In addition, using this system, we isolated a mutant of subtilisin E with higher activity by optimizing the amino acid adjacent to the catalytic triad.4 In the present study, on the basis of structural comparison with other serine proteases belonging to the subtilisin family, we attempted to enhance the thermostability5 and to alter the substrate specificity of subtilisin E.6
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