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In planta differential targeting analysis of Thermotoga maritima Cel5A and CBM6-engineered Cel5A for autohydrolysis

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Abstract

The heterologous expression of glycosyl hydrolases in bioenergy crops can improve the lignocellulosic conversion process for ethanol production. We attempted to obtain high-level expression of an intact Thermotoga maritima endoglucanase, Cel5A, and CBM6-engineered Cel5A in transgenic tobacco plants for the mass production and autohydrolysis of endoglucanase. Cel5A expression was targeted to different subcellular compartments, namely, the cytosol, apoplast, and chloroplast, using the native form of the pathogenesis-related protein 1a (PR1a) and Rubisco activase (RA) transit peptides. Cel5A transgenic tobacco plants with the chloroplast transit peptide showed the highest average endoglucanase activity and protein accumulation up to 4.5% total soluble protein. Cel5A-CBM6 was targeted to the chloroplast and accumulated up to 5.2% total soluble protein. In terms of the direct conversion of plant tissue into free sugar, the Cel5A-CBM6 transgenic plant was 33% more efficient than the Cel5A transgenic plant. The protein stability of Cel5A and Cel5A-CBM6 in lyophilized leaf material is an additional advantage in the bioconversion process.

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Acknowledgments

This work was supported by the Priority Research Centers Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (Project No. 2010-0020141) to H.-J. Bae and by the World Class University project of the Ministry of Science and Technology of Korea (R31-2009-000-20025-0). SAM is grateful for funding through the BK21 program provided by the Ministry of Education.

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Authors and Affiliations

  1. Department of Forest Products and Technology (BK21 Program), Chonnam National University, Gwangju, 500-757, Republic of Korea

    Shobana Arumugam Mahadevan, Kwang Ho Lee & Hyeun-Jong Bae

  2. Bio-energy Research Institute, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Seung Gon Wi, Yeon Ok Kim & Hyeun-Jong Bae

  3. Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Hyeun-Jong Bae

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  1. Shobana Arumugam Mahadevan
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  2. Seung Gon Wi
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  3. Yeon Ok Kim
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  4. Kwang Ho Lee
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  5. Hyeun-Jong Bae
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Correspondence to Hyeun-Jong Bae.

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Mahadevan, S.A., Wi, S.G., Kim, Y.O. et al. In planta differential targeting analysis of Thermotoga maritima Cel5A and CBM6-engineered Cel5A for autohydrolysis. Transgenic Res 20, 877–886 (2011). https://doi.org/10.1007/s11248-010-9464-8

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  • DOI: https://doi.org/10.1007/s11248-010-9464-8

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