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Molecular Cloning, Characterization, and Dye-Decolorizing Ability of a Temperature- and pH-Stable Laccase from Bacillus subtilis X1

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Abstract

Laccases from fungal origin are typically unstable at high temperatures and alkaline conditions. This characteristic limits their practical applications. In this study, a new bacterial strain exhibiting laccase activity was isolated from raw fennel honey samples and identified as Bacillus subtilis X1. The CotA-laccase gene was cloned from strain X1 and efficiently expressed in Escherichia coli in a biologically active form. The purified recombinant laccase demonstrated an extensive pH range for catalyzing substrates and high stability toward alkaline pH and high temperatures. No loss of laccase activity was observed at pH 9.0 after 10 days of incubation, and approximately 21 % of the initial activity was detected after 10 h at 80 °C. Two anthraquinonic dyes (reactive blue 4 and reactive yellow brown) and two azo dyes (reactive red 11 and reactive brilliant orange) could be partially decolorized by purified laccase in the absence of a mediator. The decolorization process was efficiently promoted when methylsyringate was present, with more than 90 % of color removal occurring in 3 h at pH 7.0 or 9.0. These unusual properties indicated a high potential of the novel CotA-laccase for industrial applications.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31101331), the Open Project Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (KLIB-KF201001), and New Teachers’ Fund for Doctor Stations, Ministry of Education (20110093120002).

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, People’s Republic of China

    Zheng-Bing Guan, Ning Zhang, Chen-Meng Song, Wen Zhou, Lin-Xi Zhou, Hong Zhao, Cheng-Wen Xu, Yu-Jie Cai & Xiang-Ru Liao

  2. School of Biotechnology, Jiangnan University, Wuxi, 214122, People’s Republic of China

    Zheng-Bing Guan, Ning Zhang, Chen-Meng Song, Wen Zhou, Lin-Xi Zhou, Hong Zhao, Cheng-Wen Xu, Yu-Jie Cai & Xiang-Ru Liao

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  1. Zheng-Bing Guan
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  2. Ning Zhang
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  4. Wen Zhou
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Correspondence to Zheng-Bing Guan or Xiang-Ru Liao.

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Guan, ZB., Zhang, N., Song, CM. et al. Molecular Cloning, Characterization, and Dye-Decolorizing Ability of a Temperature- and pH-Stable Laccase from Bacillus subtilis X1. Appl Biochem Biotechnol 172, 1147–1157 (2014). https://doi.org/10.1007/s12010-013-0614-3

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  • DOI: https://doi.org/10.1007/s12010-013-0614-3

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