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Modification and simulation of Rhizomucor miehei lipase: the influence of surficial electrostatic interaction on enantioselectivity

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Abstract

Surface residues have a significant impact on the enantioselectivity of lipases. But the molecular basis of this has never been explained. In this work, transition state complexes of Rhizomucor miehei lipase (RmL) and (R)- or (S)-n-butyl 2-phenxypropinate were studied using molecular dynamics. According to comparison between B-factor of the two simulated complexes, the β 1–β 2 loop and α 2 helix were considered the enantioselectivity-determining domains of RmL. Interaction analysis of these domains suggested an Asp61–Arg86 electrostatic interaction linking the loop and helix strongly impacting enantioselectivity of RmL. Modification of Arg86 by 1, 2-cyclohexanedione weakening this interaction decreased the E ratio from 6 to 1, modification by 1-iodo-2, 3-butanedione covalently bonding Asp61 and Arg86 strengthening the interaction increased the E ratio to 45. Dynamics simulation and energy calculation of the modified lipases also displayed corresponding decreases or increases of enantioselectivity.

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Acknowledgments

This work was financially supported by the National Basic Research Program of China (973 Program, No. 2011CB710800), Hi-Tech Research and Development Program of China (863 Program, 2011AA02A209), National Natural Science Foundation of China (No. 20936002), and the Science and Technology Planning Project of Zhejiang Province (No. 2010C31127).

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    Authors and Affiliations

    1. Department of Chemical and Biological Engineering, Institute of Biological Engineering, Zhejiang University, Hangzhou, 310027, China

      Gang Xu, Xiao Meng, Lin-Jie Xu, Li Guo, Jian-Ping Wu & Li-Rong Yang

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    1. Gang Xu
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    2. Xiao Meng
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    3. Lin-Jie Xu
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    4. Li Guo
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    Correspondence to Jian-Ping Wu.

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    Gang Xu and Xiao Meng have contributed equally to this work.

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    Xu, G., Meng, X., Xu, LJ. et al. Modification and simulation of Rhizomucor miehei lipase: the influence of surficial electrostatic interaction on enantioselectivity. Biotechnol Lett 37, 871–880 (2015). https://doi.org/10.1007/s10529-014-1747-3

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