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Heterologous expression and characterisation of a laccase from Colletotrichum lagenarium and decolourisation of different synthetic dyes

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Abstract

Laccases have received considerable attention in recent decades because of their ability to oxidise a large spectrum of phenolic and non-phenolic organic substrates and highly recalcitrant environmental pollutants. In this research, a laccase gene from Colletotrichum lagenarium was chemically synthesised using yeast bias codons and expressed in Pichia pastoris. The molecular mass of the recombinant laccase was estimated to be 64.6 kDa by SDS–PAGE, and the enzyme exhibited maximum activity at pH 3.6–4.0 but more stability in buffer with higher pH (>pH 3.6). The optimal reaction temperature of the enzyme was 40 °C, beyond which stability significantly decreased. By using 2,2′-azino-bis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS) as a substrate, K m and V max values of 0.34 mM and 7.11 mM min−1 mg−1, respectively, were obtained. Using ABTS as a mediator, the laccase could oxidise hydroquinone to p-benzoquinone and decolourise the synthetic dyes malachite green, crystal violet and orange G. These results indicated that the laccase could be used to treat industrial effluents containing artificial dyes.

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Acknowledgments

This research was supported by the Key Project Fund of the Shanghai Municipal Committee of Agriculture (Grant Nos. zhongzi2013-8, zhongzi2014-2, gongzi2014 7-1-3), Agriculture Science Technology Achievements Transformation Fund (Grant No. 143919N0300), National Natural Science Foundation of China (Grant Nos. 31071486, 31200212, 31200075 and 31200076), Basic Research in the Field of Science and Technology Project of Science and Technology Commission of Shanghai Municipality (Grant No. 14JC1403602) and The Growth Plan for the Young from Shanghai Municipal Committee of Agriculture. The funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript.

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Authors and Affiliations

  1. Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China

    Bo Wang, Yongsheng Tian, Wei Zhao, Zhengjun Li, Jianjie Gao, Rihe Peng & Quanhong Yao

  2. Crop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201403, China

    Ying Yan

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  1. Bo Wang
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  2. Ying Yan
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  3. Yongsheng Tian
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  4. Wei Zhao
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Correspondence to Quanhong Yao.

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Wang, B., Yan, Y., Tian, Y. et al. Heterologous expression and characterisation of a laccase from Colletotrichum lagenarium and decolourisation of different synthetic dyes. World J Microbiol Biotechnol 32, 40 (2016). https://doi.org/10.1007/s11274-015-1999-7

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  • DOI: https://doi.org/10.1007/s11274-015-1999-7

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