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Applied Microbiology and Biotechnology

, Volume 98, Issue 15, pp 6679–6687 | Cite as

Characterization of a multi-function processive endoglucanase CHU_2103 from Cytophaga hutchinsonii

  • Cong Zhang
  • Ying Wang
  • Zhe Li
  • Xiangru Zhou
  • Weican Zhang
  • Yue Zhao
  • Xuemei LuEmail author
Biotechnologically relevant enzymes and proteins

Abstract

Cytophaga hutchinsonii is a Gram-negative gliding bacterium which can efficiently degrade crystalline cellulose by an unknown strategy. Genomic analysis suggests the C. hutchinsonii genome lacks homologs to an obvious exoglucanase that previously seemed essential for cellulose degradation. One of the putative endoglucanases, CHU_2103, was successfully expressed in Escherichia coli JM109 and identified as a processive endoglucanase with transglycosylation activity. It could hydrolyze carboxymethyl cellulose (CMC) into cellodextrins and rapidly decrease the viscosity of CMC. When regenerated amorphous cellulose (RAC) was degraded by CHU_2103, the ratio of the soluble to insoluble reducing sugars was 3.72 after 3 h with cellobiose and cellotriose as the main products, indicating that CHU_2103 was a processive endoglucanase. CHU_2103 could degrade cellodextrins of degree of polymerization ≥3. It hydrolyzed p-nitrophenyl β-d-cellodextrins by cutting glucose or cellobiose from the non-reducing end. Meanwhile, some larger-molecular-weight cellodextrins could be detected, indicating it also had transglycosylation activity. Without carbohydrate-binding module (CBM), CHU_2103 could bind to crystalline cellulose and acted processively on it. Site-directed mutation of CHU_2103 demonstrated that the conserved aromatic amino acid W197 in the catalytic domain was essential not only for its processive activity, but also its cellulose binding ability.

Keywords

Cytophaga hutchinsonii Family 5 glycoside hydrolase Processive endoglucanase Multi-function 

Notes

Acknowledgements

This work was supported by the National Basic Research Program of China (2011CB707402) and the National Natural Science Foundation of China (31170051 and 31371262). We sincerely thank Dr. Mark J. McBride (University of Wisconsin–Milwaukee, Milwaukee, WI, USA) for providing C. hutchinsonii ATCC 33406. Thanks to Dr. Edward C. Mignot, Shandong University, for linguistic advice.

Supplementary material

253_2014_5640_MOESM1_ESM.pdf (325 kb)
ESM 1 (PDF 324 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Cong Zhang
    • 1
  • Ying Wang
    • 1
  • Zhe Li
    • 1
  • Xiangru Zhou
    • 1
  • Weican Zhang
    • 1
  • Yue Zhao
    • 1
  • Xuemei Lu
    • 1
    Email author
  1. 1.State Key Laboratory of Microbial Technology, School of Life ScienceShandong UniversityJinanChina

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