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Exploring antibiotic resistance based on enzyme hydrolysis by microcalorimetry

Part IV. Determination of thermokinetic parameters of D-Ala-D-Ala hydrolysis with dipeptidase VanX

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Abstract

In an effort to explore antibiotic resistance based on enzyme hydrolysis, the thermodynamic parameters of the D-Ala-D-Ala hydrolysis catalyzed by dipeptidase VanX and occurred in Gram-positive vancomycin-resistant pathogens were determined by microcalorimetry. The values of activation free energy \( \Updelta G_{ \ne }^{\theta } \) are 87.140 ± 0.055, 88.413 ± 0.067, 89.611 ± 0.051, and 90.823 ± 0.042 kJ mol−1 at 293.15, 298.15, 303.15, and 308.15 K, respectively, activation enthalpy \( \Updelta H_{ \ne }^{\theta } \) is 15.332 ± 0.006 kJ mol−1, activation entropy \( \Updelta S_{ \ne }^{\theta } \) is −245.02 ± 0.20 J mol−1 K−1, apparent activation energy E is 17.830 kJ mol−1, and the reaction order is 1.5. These thermodynamic data reveal that D-Ala-D-Ala hydrolysis with VanX is an exothermic and spontaneous reaction and has an approximative reaction rate with the imipenem hydrolysis with metallo-β-lactamase ImiS in vitro.

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Acknowledgements

We are grateful to Professor Michael Crowder at Miami University for providing the plasmid pIADL14 for expression of MBP-VanX. This study was supported by grants (to K. W. Y) from National Natural Science Fund of China (20972127), Doctoral Fund of China (200806970005), Natural Science Fund of Shaanxi Province (2009JM2002), and Key Fund for International Cooperation of Shaanxi Province (2010KW-16).

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Authors and Affiliations

  1. Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi’an, 710069, People’s Republic of China

    Cheng-Cheng Liu, Xiang-Bo Zhao, Ke-Wu Yang, Le Zhai, Xia Yang & Hui-Zhou Gao

  2. School of Chemical Engineering, Northwest University, Xi’an, 710069, People’s Republic of China

    Kang-Zhen Xu

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  1. Cheng-Cheng Liu
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  2. Xiang-Bo Zhao
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Correspondence to Ke-Wu Yang.

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Liu, CC., Zhao, XB., Yang, KW. et al. Exploring antibiotic resistance based on enzyme hydrolysis by microcalorimetry. J Therm Anal Calorim 111, 1663–1667 (2013). https://doi.org/10.1007/s10973-011-2010-y

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