Towards understanding directed evolution: more than half of all amino acid positions contribute to ionic liquid resistance of Bacillus subtilis lipase A

Chembiochem : a European Journal of Chemical Biology
Victorine Josiane Frauenkron-MachedjouUlrich Schwaneberg

Abstract

Ionic liquids (ILs) are attractive (co-)solvents for biocatalysis. However, in high concentration (>10 % IL), enzymes usually show decreased activity. No general principles have been discovered to improve IL resistance of enzymes by protein engineering. We present a systematic study to elucidate general engineering principles by site saturation mutagenesis on the complete gene bsla. Screening in presence of four [BMIM]-based ILs revealed two unexpected lessons on directed evolution: 1) resistance improvement was obtainable at 50-69 % of all amino acid positions, thus explaining the success of small sized random mutant libraries; 2) 6-13 % of substitutions led to improved resistance. Among these, 66-95 % were substitutions by chemically different amino acids (e.g., aromatic to polar/aliphatic/charged amino acids), thus indicating that mutagenesis methods introducing such changes should, at least for lipases like BSLA, be favored to improve IL resistance.

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Citations

Jan 29, 2016·Protein Engineering, Design & Selection : PEDS·Paul W WolskiHarvey W Blanch
Sep 22, 2015·Chembiochem : a European Journal of Chemical Biology·Erik M NordwaldJoel L Kaar
Apr 16, 2015·Chemical Communications : Chem Comm·Feng ChengUlrich Schwaneberg
May 18, 2016·Scientific Reports·Georgette KörferUlrich Schwaneberg
May 4, 2017·Biotechnology and Bioengineering·Jianhua YangUlrich Schwaneberg
Jan 5, 2017·Applied Microbiology and Biotechnology·Jing ZhaoUlrich Schwaneberg
Sep 29, 2018·Molecular Biotechnology·Angelica Jimenez-Rosales, Miriam V Flores-Merino
Nov 14, 2020·Chemistry : a European Journal·Haiyang CuiUlrich Schwaneberg
Dec 29, 2020·International Journal of Biological Macromolecules·Karen Rodríguez-NúñezRonny Martínez
Jan 7, 2020·Journal of Chemical Information and Modeling·Christina NutschelHolger Gohlke
Aug 28, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Hasan Tanvir ImamAndrew Craig Marr
Aug 26, 2021·Computational and Structural Biotechnology Journal·Till El HarrarHolger Gohlke

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