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. 2022 Dec 29;27(6):930–937. doi: 10.1007/s12257-022-0317-0

Decolorization of Textile Dye by Spore Surface Displayed Small Laccase for the Enhanced Thermal Stability and Robust Repeated Reaction

Wooil Kim 1,#, Yura Jeong 1,2,#, Sanggeun Back 1, Suhwan Kim 1,2, Junehyung Kim 1,3,✉
PMCID: PMC9798364  PMID: 36593775

Abstract

In this study, we tried to decolorize synthetic dyes using small laccase (SLAC) from Streptomyces coelicolor, which is resistant to pH, temperature change, and traditional inhibitors for the actual industrial applications using spore surface display system. We inserted SLAC-His6 tag at the C-terminal of CotE anchoring motif. The proper surface expression of CotE-SLAC fusion protein on the surface of Bacillus subtilis spore was verified with flow cytometry using FITC labeled anti-His6 tag antibody. After 6 h of reaction, more than 90% of Indigo carmine was decomposed using recombinant SLAC displaying Bacillus spore, whereas less than 10% of Indigo carmine was decomposed with wild type spore. Over 70% of laccase activity was retained with recombinant SLAC displaying spore, which was heat-treated for 3 h at 90°C. For eight rounds of repeated decomposition of Indigo carmine, no significant decrease of enzymatic activity was observed. This showed the robust characteristics of spore display format for repeated and harsh condition reactions.

Keywords: Bacillus subtilis, spore display, small laccase, decolorization

Acknowledgements

This research was supported by a grant (NRF-2017R1D1A 1B0503572614) of the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Republic of Korea.

Ethical Statements

The authors declare no conflict of interest.

Neither ethical approval nor informed consent was required for this study.

Footnotes

Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Wooil Kim and Yura Jeong contributed equally to this work.

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