A Single Ser 85 Ala Mutation enhances the catalytic efficiency of Subtilisin E from Bacillus subtilis 168. | [Biotechnology • 2004]

Author(s):

1. Eliel R. Romerio-Garcia: Institute of Investigation in Experimental Biology, Faculty of Chemistry, University of Guanajuato, Noria Alta, S/N.Guanajuato, 36050, Mexico

2. Joel A. Esquivel-Naranho: Institute of Investigation in Experimental Biology, Faculty of Chemistry, University of Guanajuato, Noria Alta, S/N.Guanajuato, 36050, Mexico

3. Norma Ramirez Ramirez: Institute of Investigation in Experimental Biology, Faculty of Chemistry, University of Guanajuato, Noria Alta, S/N.Guanajuato, 36050, Mexico

4. Jesus Garcia-Soto: Institute of Investigation in Experimental Biology, Faculty of Chemistry, University of Guanajuato, Noria Alta, S/N.Guanajuato, 36050, Mexico

5. Mario Pedraza¬Reyes: Institute of Investigation in Experimental Biology, Faculty of Chemistry, University of Guanajuato, Noria Alta, S/N.Guanajuato, 36050, Mexico

Abstract:

A single mutation of Serbr Ala was. introduced into the highly conserved 83-Gly-Val-Ala-85 region of AprE (subtilisin E), which resulted in the generation of a subtilisin mutant with an enhanced catalytic efficiency. The position of the mutation was placed in the conserved N-terminal end of the loop that connects a (3-sheet with the (x-helix containing the catalytic residue His64.The mutant Ser 85 Ala aprE gene was over expressed in a protease deficient B. subtilius genetic background and its product purified to homogeneity. The Ser85 Ala AprE protein exhibited a catalytic efficiency two fold higher than that of its wild type parent AprE due to a larger kc,,. Other biochemical properties exhibited by the pure wild type subtilisin. These results support the idea that mutations on the conserved stretch bend 83-Gly-Val-Ser85, which connects two elements of secondary structure in AprE cause alterations on the catalytic properties of AprE and other subtilisins.

Page(s): 49-55

DOI: DOI not available

Published: Journal: Biotechnology, Volume: 3, Issue: 1, Year: 2004

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