Identification of amino acid residues essential for the catalytic reaction of Bacillus kaustophilus leucine aminopeptidase |
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Authors: | Chi Meng-Chun Chou Wei-Mou Hsu Wen-Hwei Lin Long-Liu |
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Institution: | Graduate Institute of Biotechnology, National Chiayi University, 60083 Chiayi, Taiwan. |
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Abstract: | The functional significance of amino acid residues Lys-265, Asp-270, Lys-277, Asp-288, Asp-347, Glu-349, and Arg-351 of Bacillus kaustophilus leucine aminopeptidase was explored by site-directed mutagenesis. Variants with an apparent molecular mass of approximately 54 kDa were overexpressed in Escherichia coli and purified to homogeneity by nickel-chelate chromatography. The purified mutant enzymes had no LAP activity, implying that these residues are important for the catalytic reaction of the enzyme. |
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