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Cloning of Thermostable Cellulase Genes of Clostridium thermocellum and Their Secretive Expression in Bacillus subtilis

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Abstract

Screening for the powerful cellulase genes with improved activities remains a challenge for the biorefinery research. In this study, five cellobiohydrolase genes and one endoglucanase gene sourced from Clostridium thermocellum DSM 1237, cbhA, celK, celO, cel48Y, cel48S, and celA were cloned into a newly established tool vector pP43JM2 and expressed in two Bacillus subtilis strains, B. subtilis WB600 and B. subtilis WB800, respectively. Most of the cellulases produced in the B. subtilis recombinants were efficiently secreted into the culture medium. These secreted soluble proteins showed distinct cellulase activities using phosphoric acid swollen cellulose (PASC) as the substrate and they also demonstrated strong synergistic effects for PASC, Avicel cellulose, and the dilute acid pretreated corn stover. The current work provided a quick secretive cloning method for screening cellulase genes and may provide a host strain for constructing a consolidated bioprocessing platform with the capacity of secretive expression of multiple cellulases.

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Acknowledgments

This research was supported by National Basic Research Program of China (2011CB707406), National Natural Science Foundation of China (20976051), Ministry of Education of China (107123, 20090074110013), Shanghai Leading Academic Discipline Project (B505), Fundamental Research Funds for the Central Universities of China (WF0913005), and China National Special Funds for the State Key Laboratory of Bioreactor Engineering (2060204).

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

    Jian-Ming Liu, Xiu-Juan Xin, Chun-Xiu Li, Jian-He Xu & Jie Bao

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  1. Jian-Ming Liu
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  2. Xiu-Juan Xin
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  3. Chun-Xiu Li
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Correspondence to Jie Bao.

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Liu, JM., Xin, XJ., Li, CX. et al. Cloning of Thermostable Cellulase Genes of Clostridium thermocellum and Their Secretive Expression in Bacillus subtilis . Appl Biochem Biotechnol 166, 652–662 (2012). https://doi.org/10.1007/s12010-011-9456-z

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