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A useful method integrating production and immobilization of recombinant cellulase

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Abstract

The development of cellulase-based bioprocess is afflicted by the processing efficiency of enzymes. To address this issue, a method based on artificial oil bodies (AOBs) was proposed to integrate production and immobilization of recombinant cellulase. First, the heterologous endoglucanase (celA), cellobiohydrolase (celK), and β-glucosidase (gls) genes were individually fused with oleosin, a structural protein of plant seed oils. After expression in Escherichia coli, each fusion protein of insolubility was mixed together with plant oils. AOBs were assembled by subjecting the mixture to sonication. Consequently, active CelA, CelK, and Gls were resumed and co-immobilized on AOBs surface. Finally, the assembly condition (including the protein ratio) and the reaction condition were further optimized by response surface methodology. The resulting AOBs-bound cellulase remained stable for 4 cycles of cellulose–hydrolyzed reactions. Overall, the result shows a promise of this proposed approach for processing recombinant cellulase, which may provide a facile method to investigate optimum combination of cellulase components towards various cellulosic materials.

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Acknowledgments

This work is supported by the National Science Council of Taiwan (NSC98-3114 -E-035-001-CC2; NSC102-2622-E-035-005-CC1; NSC101-2221-E-035-057-MY3) and Ministry of Economic Affairs (101-EC-17-A-10-S1-156).

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Authors and Affiliations

  1. Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 40402, Taiwan

    Chung-Jen Chiang

  2. Department of Biotechnology, Southern Taiwan University of Science and Technology, No. 1, Nantai St., Tainan, 71005, Taiwan

    Po Ting Chen

  3. Department of Chemical Engineering, Feng Chia University, 100 Wen-Hwa Road, Taichung, 40724, Taiwan

    Chien Yu Yeh, Zei Wen Wang & Yun-Peng Chao

  4. Department of Health and Nutrition Biotechnology, Asia University, Taichung, 41354, Taiwan

    Yun-Peng Chao

  5. Department of Medical Research, China Medical University Hospital, Taichung, 40447, Taiwan

    Yun-Peng Chao

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  1. Chung-Jen Chiang
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  2. Po Ting Chen
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  3. Chien Yu Yeh
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  4. Zei Wen Wang
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  5. Yun-Peng Chao
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Correspondence to Yun-Peng Chao.

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Chiang, CJ., Chen, P.T., Yeh, C.Y. et al. A useful method integrating production and immobilization of recombinant cellulase. Appl Microbiol Biotechnol 97, 9185–9192 (2013). https://doi.org/10.1007/s00253-013-5238-6

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  • DOI: https://doi.org/10.1007/s00253-013-5238-6

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