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

GUAN Zhengbing; ZHANG Ning; SONG Chenmeng; LIAO Xiangru

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

GUAN Zhengbing * #,ZHANG Ning,SONG Chenmeng,LIAO Xiangru

Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, P. R. China

*Correspondence author

#Submitted by

Subject:

Funding: none

Opened online:23 October 2013

Accepted by: none

Citation: GUAN Zhengbing,ZHANG Ning,SONG Chenmeng.Molecular Cloning, Characterization, and Dye-Decolorizing Ability of a Laccase from Bacillus subtilis X1[OL]. [23 October 2013] http://en.paper.edu.cn/en_releasepaper/content/4564243

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 d 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 pH 9.0. These unusual properties indicated a high potential of the novel CotA-laccase for industrial applications.

Keywords:Microbiology; Laccase; Bacillus subtilis; Gene cloning; Recombinant expression; Dye decolorization?

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