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Role of the Conserved Thr399 and Thr417 Residues of Bacillus licheniformis γ-Glutamyltranspeptidase as Evaluated by Mutational Analysis

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An Erratum to this article was published on 06 October 2009

An Erratum to this article was published on 06 October 2009

Abstract

Role of the conserved Thr399 and Thr417 residues of Bacillus licheniformis γ-glutamyltranspeptidase (BlGGT) was investigated by site-directed mutagenesis. Substitutions of Thr399 and Thr417 of BlGGT with Ser resulted in a dramatic reduction in enzymatic activity. A complete loss of the GGT activity was observed in T399A, T399C, T417A, and T417K mutant enzymes. Furthermore, mutations on these two residues impaired the capability of autocatalytic processing of the enzyme. In vitro maturation experiments showed that BlGGT mutant precursors, pro-T399S, pro-T417S, and pro-T417A, could precede a time-dependent autocatalytic process to generate the 44.9- and 21.7-kDa subunits; however, the processed T417A had no enzymatic activity. Measurement of intrinsic tryptophan fluorescence revealed alteration of the microenvironment of aromatic amino acid residues, while Far-UV circular dichroism spectra were nearly identical for wild-type and mutant enzymes. These results suggest that residues Thr399 and Thr417 are important for BlGGT in the enzymatic maturation and reaction.

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Acknowledgments

The authors are grateful to the National Science Council of Taiwan for financial supports (NSC 95-2313-B-415-012-MY3 and NSC 97-2628-B-415-001-MY3.

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Authors and Affiliations

  1. Department of Applied Chemistry, National Chiayi University, 300 University Road, Chiayi, 600, Taiwan

    Rui-Cin Lyu, Hui-Ping Chang & Long-Liu Lin

  2. Department of Food and Nutrition, Hungkuang University, Shalu, Taichung, Taiwan

    Hui-Yu Hu, Lih-Ying Kuo & Huei-Fen Lo

  3. Department of Biochemical Science and Technology, National Chiayi University, 300 University Road, Chiayi, 600, Taiwan

    Ping-Lin Ong

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  1. Rui-Cin Lyu
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  2. Hui-Yu Hu
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Correspondence to Long-Liu Lin.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s00284-009-9509-5

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Lyu, RC., Hu, HY., Kuo, LY. et al. Role of the Conserved Thr399 and Thr417 Residues of Bacillus licheniformis γ-Glutamyltranspeptidase as Evaluated by Mutational Analysis. Curr Microbiol 59, 101–106 (2009). https://doi.org/10.1007/s00284-009-9403-1

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  • DOI: https://doi.org/10.1007/s00284-009-9403-1

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