Abstract
Laccase (benzenediol: oxygen oxidoreductases, EC1.10.3.2) is a multi-copper oxidase capable of oxidizing a variety of phenolic and other aromatic organic compounds. The catalytic power of laccase makes it an attractive candidate for potential applications in many areas of industry including biodegradation of organic pollutants and synthesis of novel drugs. Most laccases are vulnerable to high salt and have limited applications. However, some laccases are not only tolerant to but also activated by certain concentrations of salt and thus have great application potential. The mechanisms of salt-induced activity enhancement of laccases are unclear as yet. In this study, we used dynamic light scattering, size exclusion chromatography, analytical ultracentrifugation, intrinsic fluorescence emission, circular dichroism, ultraviolet–visible light absorption, and an enzymatic assay to investigate the potential correlation between the structure and activity of the marine-derived laccase, Lac15, whose activity is promoted by low concentrations of NaCl. The results showed that low concentrations of NaCl exert little influence on the protein structure, which was partially folded in the absence of the salt; moreover, the partially folded rather than the fully folded state seemed to be favorable for enzyme activity, and this partially folded state was distinctive from the so-called ‘molten globule’ occasionally observed in active enzymes. More data indicated that salt might promote laccase activity through mechanisms involving perturbation of specific local sites rather than a change in global structure. Potential binding sites for chloride ions and their roles in enzyme activity promotion are proposed.
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Abbreviations
- 2,6-DMP:
-
2,6-Dimethoxyphenol
- ABTS:
-
2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)
- CD:
-
Circular dichroism
- DLS:
-
Dynamic light scattering
- SEC:
-
Size exclusion chromatography
- SDS-PAGE:
-
Sodium dodecyl sulfate polyacrylamide gel electrophoresis
- SGZ:
-
Syringaldazine
- TFE:
-
Trifluoroethanol
- TMAO:
-
Trimethylamine N-oxide
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Acknowledgements
We thank Qian Wang at the Institute of Microbiology of the Chinese Academy of Sciences for help in analyzing the AUC data. This work was supported by grants from the National Natural Science Foundations of China (31370114 to Y.X and 31470775 to X.Z), the National High Technology Research and Development Program of China (2011AA09070305 to Y.X), the International Science and Technology Cooperation Plan of Anhui Province (1503062010 to X.Z) and the Doctoral Fund of the Ministry of Education of China (20133401110006 to Y.X).
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Li, J., Xie, Y., Wang, R. et al. Mechanism of salt-induced activity enhancement of a marine-derived laccase, Lac15. Eur Biophys J 47, 225–236 (2018). https://doi.org/10.1007/s00249-017-1251-5
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DOI: https://doi.org/10.1007/s00249-017-1251-5