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Chemical modification for improving catalytic performance of lipase B from Candida antarctica with hydrophobic proline ionic liquid

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Abstract

In this study, a series of proline ionic liquids with different lengths of hydrophobic alkyl on the side chain were used to modify the Candida Antarctic lipase B (CALB). The catalytic activity, thermal stability and tolerance to methanol and DMSO of the modified enzyme were all improved simultaneously. The optimum temperature changed from 55 to 60 ℃. The hydrophobicity and anion type of the modifier have important influence on the catalytic performance of CALB. CALB modified by [ProC12][H2PO4] has a better effect. Under the optimal conditions, its hydrolysis activity was 3.0 times than that of the native enzyme, the catalytic efficiency Kcat/Km improved 2.8 times in aqueous phase, and the tolerance to organic solvent with strong polarity (50% methanol 2 h) was increased by 6.8 times. Fluorescence spectra and circular dichroism (CD) spectroscopy showed that the introduction of ionic liquids changed the microenvironment near the fluorophores of the enzyme protein, the α-helix decreased and β-sheet increased in the secondary structure of the modified enzymes. The root mean square deviation (RMSD), residue root mean square fluctuation (RMSF), radius of gyration (Rg), and solution accessible surface area (SASA) of [ProC2][Br]-CALB, [ProC12][Br]-CALB and native CALB were obtained for comparison by molecular dynamics simulation. The results of dynamics simulation were in good agreement with enzymology experiment. The introduction of ionic liquids can keep CALB in a better active conformation, and proline ionic liquids with long hydrophobic chains can significantly improve the surface hydrophobicity and overall rigidity of CALB. This research offers a new idea for rapid screening of efficient modifiers and provision of enzymes with high stability and activity for industrial application.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China. (22178174, 21676143).

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  1. State Key Laboratory of Materials-Oriented Chemical Engineering, School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing, 210009, China

    Xiao-guang Zhang, Yu Xue, Ze-ping Lu, Hua-jin Xu & Yi Hu

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  1. Xiao-guang Zhang
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Zhang, Xg., Xue, Y., Lu, Zp. et al. Chemical modification for improving catalytic performance of lipase B from Candida antarctica with hydrophobic proline ionic liquid. Bioprocess Biosyst Eng 45, 749–759 (2022). https://doi.org/10.1007/s00449-022-02696-x

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