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Heterologous Expression Characteristics of Trichoderma viride Endoglucanase V in the Silkworm, Bombyx mori L.

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Abstract

Efficient degradation of cellulose needs a synergistic reaction of the cellulolytic enzymes, which include exoglucanases, endoglucanases, and β-1,4-glucosidase. In this study, we used an improved Bac-to-Bac/BmNPV baculovirus expression system, which lacks the virus-encoded chitinase cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV), to express the endoglucanase V (EG V) gene from Trichoderma viride in silkworm BmN cells and silkworm larvae, and analyzed the characteristics of the recombinant enzyme in silkworm larvae. The result showed that an around 36-kDa protein was visualized in BmN cells at 48 h after the second-generation recombinant mBacmid/BmNPV/EG V baculovirus infection. The crude enzyme extract from the recombinant baculoviruses-infected silkworms exhibited a significant maximum activity at the environmental condition of pH 5.0 and a temperature of 50 °C, and increased 39.86% and 37.76% compared with that from blank mBacmid/BmNPV baculovirus-infected silkworms and normal silkworms, respectively. It was stable at pH range from 5.0 to 10.0 and at temperature range from 40 to 60 °C. The availability of large quantities of EG V that the silkworm provides might greatly facilitate the future research and the potential application in industries.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (no. 30972141/C120110) and the Chinese Universities Scientific Fund.

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Authors and Affiliations

  1. Key Laboratory of Animal Epidemic Etiology and Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Xing-hua Li, Mei-xian Wang, Peng Zhang, Jia-biao Hu, Chun-guang Sun, Fang Zhou, Yan-shan Niu, Firdose Ahmad Malik, Roy Bhaskar, Hua-jun Yang & Yun-gen Miao

  2. Sericultural Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China

    Xin-ju Liu

  3. College of Animal Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Yun-gen Miao

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  1. Xing-hua Li
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  2. Mei-xian Wang
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  3. Peng Zhang
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  4. Jia-biao Hu
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  5. Chun-guang Sun
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  6. Xin-ju Liu
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  7. Fang Zhou
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  8. Yan-shan Niu
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  9. Firdose Ahmad Malik
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  10. Roy Bhaskar
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  11. Hua-jun Yang
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  12. Yun-gen Miao
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Correspondence to Yun-gen Miao.

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Li, Xh., Wang, Mx., Zhang, P. et al. Heterologous Expression Characteristics of Trichoderma viride Endoglucanase V in the Silkworm, Bombyx mori L.. Appl Biochem Biotechnol 165, 728–736 (2011). https://doi.org/10.1007/s12010-011-9291-2

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  • DOI: https://doi.org/10.1007/s12010-011-9291-2

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