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Hydrophobic Substitution of Surface Residues Affects Lipase Stability in Organic Solvents

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Abstract

A novel lipase has been recently isolated from a local Pseudomonas sp. (GQ243724). In the present study, we have tried to increase the organic solvent stability of this lipase using site-directed mutagenesis. Eight variants N219L, N219I, N219P, N219A, N219R, N219D, S251L, and S251K were designed to change the surface hydrophobicity of this enzyme with respect to the wild-type. Among these variants, the stability of N219L and N219I significantly increased in the presence of all tested organic solvents, whereas two mutants (N219R and N219D) significantly exhibited decreased stabilities in all the organic solvent studied, suggesting that improvement of hydrophobic patches on the enzyme surface enhances the stability in organic media. Furthermore, replacing Ser251 with hydrophobic residues on the enzyme surface dramatically diminished its stability in the tested condition. In spite of the distance of the mutated sites from the active site, the values of k cat and K m were affected. Finally, structural analysis of the wild-type and mutated variants was carried out in the presence and absence of some organic solvents using circular dichroism and fluorescence spectroscopy.

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Acknowledgments

The authors would like to express their gratitude to the Research Council of the University of Tehran and Tarbiat Modares University for their financial support.

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Authors and Affiliations

  1. Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran

    Maryam Monsef Shokri & Shahin Ahmadian

  2. Department of Microbiology, Faculty of Science, Islamic Azad University, Arak branch, Arak, Iran

    Neda Akbari

  3. Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

    Khosro Khajeh

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  1. Maryam Monsef Shokri
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  2. Shahin Ahmadian
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Correspondence to Shahin Ahmadian or Khosro Khajeh.

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Monsef Shokri, M., Ahmadian, S., Akbari, N. et al. Hydrophobic Substitution of Surface Residues Affects Lipase Stability in Organic Solvents. Mol Biotechnol 56, 360–368 (2014). https://doi.org/10.1007/s12033-013-9716-y

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