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Expression and Characterization of a Recombinant Laccase with Alkalistable and Thermostable Properties from Streptomyces griseorubens JSD-1

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Abstract

Streptomyces griseorubens JSD-1 is a novel actinomycete that could grow efficiently upon lignin, and the ligninolytic genes active in this biotransformation were expected to be crucial. To investigate the molecular mechanism of utilizing lignin, genome sequencing was carried out to obtain its draft genome, which was deposited at GenBank under the accession No. JJMG00000000. Multiple copper oxidase (MCO) was obtained, which proved to be an extracellular enzyme and have relative high expression with the stimulation of ligninolytic materials. Judging from its putative 3D structure, the N-terminal of MCO was bared, which was fit for the linkage of poly-HIS10 tag. As a result, heterogeneous expression conditions of recombinant laccase was achieved with TransB(DE3) grown in a modified terrific broth (TB) medium with an extra addition of 0.5 % glucose at 30 °C until optical density at 600 nm (OD600) reached 0.8 when expression was induced by 25 μM isopropyl β-D-1-thiogalactopyranoside (IPTG) and also 100 μM copper sulphate as supplement. Finally, it exhibited special characters of thermal robustness, alkaline activity profiles, high resistance to metallic ions and chemical inhibitors as well as dye decolourization. In summary, our findings illustrated the genetic basic of utilizing lignin in this isolate. Additionally, a novel laccase expected to be potential in agricultural and industrial application was expressed and characterized as well.

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Acknowledgments

The research work was supported by the National High Technology Research and Development Program of China (2012AA101405), Special Fund for Agro-scientific Research in the Public Interest of China (200903056), the National Natural Science Foundation of China (20977062) and the Municipal Natural Science Foundation of Shanghai, China (13ZR1421700). We also give thanks for the sequencing service provided by Personal Biotechnology Co., Ltd. Shanghai, China.

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Authors and Affiliations

  1. School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Haiwei Feng, Dan Zhang, Yujing Sun, Yuee Zhi, Liang Mao, Yanqing Luo, Lurong Xu, Lumei Wang & Pei Zhou

  2. Key Laboratory of Urban Agriculture (South), Ministry of Agriculture, Shanghai Jiao Tong University, Shanghai, 200240, China

    Haiwei Feng, Dan Zhang, Yuee Zhi, Liang Mao, Yanqing Luo, Lurong Xu, Lumei Wang & Pei Zhou

  3. Bor S. Luh Food Safety Research Center, Shanghai Jiao Tong University, Shanghai, 200240, China

    Haiwei Feng, Dan Zhang, Yuee Zhi, Liang Mao, Yanqing Luo, Lurong Xu, Lumei Wang & Pei Zhou

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  1. Haiwei Feng
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Correspondence to Pei Zhou.

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Fig. S1

Cellular localization of MCO. (PNG 27 kb)

Fig. S2

Images of putative 3D structure modelings. (a) MCO; (b) recombinant MCO with HIS10·tag on the N-terminal. (PNG 400 kb)

Fig. S3

Comparison of codon usage through optimization. (a) MCO; (b) CO-MCO. (JPEG 346 kb)

Fig. S4

Multiple sequences alignment of MCO and CO-MCO. (JPEG 2305 kb)

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Feng, H., Zhang, D., Sun, Y. et al. Expression and Characterization of a Recombinant Laccase with Alkalistable and Thermostable Properties from Streptomyces griseorubens JSD-1. Appl Biochem Biotechnol 176, 547–562 (2015). https://doi.org/10.1007/s12010-015-1594-2

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