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Expression and characterization of a glucose-tolerant β-1,4-glucosidase with wide substrate specificity from Cytophaga hutchinsonii

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Abstract

Cytophaga hutchinsonii is a gram-negative bacterium that can efficiently degrade crystalline cellulose by a novel strategy without cell-free cellulases or cellulosomes. Genomic analysis implied that C. hutchinsonii had endoglucanases and β-glucosidases but no exoglucanases which could processively digest cellulose and produce cellobiose. In this study, BglA was functionally expressed in Escherichia coli and found to be a β-glucosidase with wide substrate specificity. It can hydrolyze pNPG, pNPC, cellobiose, and cellodextrins. Moreover, unlike most β-glucosidases whose activity greatly decreases with increasing length of the substrate chains, BglA has similar activity on cellobiose and larger cellodextrins. The K m values of BglA on cellobiose, cellotriose, and cellotetraose were calculated to be 4.8 × 10−2, 5.6 × 10−2, and 5.3 × 10−2 mol/l, respectively. These properties give BglA a great advantage to cooperate with endoglucanases in C. hutchinsonii in cellulose degradation. We proposed that C. hutchinsonii could utilize a simple cellulase system which consists of endoglucanases and β-glucosidases to completely digest amorphous cellulose into glucose. Moreover, BglA was also found to be highly tolerant to glucose as it retained 40 % activity when the concentration of glucose was 100 times higher than that of the substrate, showing potential application in the bioenergy industry.

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Acknowledgments

We sincerely thank Dr. Mark J. McBride (University of Wisconsin-Milwaukee, Milwaukee, USA) for providing C. hutchinsonii ATCC 33406 and Dr. Edward C. Mignot, Shandong University, for linguistic advice.

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Authors and Affiliations

  1. State Key Laboratory of Microbial Technology, College of Life Science, Shandong University, Jinan, 250100, China

    Cong Zhang, Xifeng Wang, Weican Zhang, Yue Zhao & Xuemei Lu

  2. Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, Xiamen, 361005, China

    Cong Zhang

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  1. Cong Zhang
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Correspondence to Xuemei Lu.

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This work was funded by the National Science Foundation of China (31371262 and 31170051) and Sci-Tech Development Project of Shandong Province (2014GGH202001).

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All authors declare that they do not have conflicts of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Zhang, C., Wang, X., Zhang, W. et al. Expression and characterization of a glucose-tolerant β-1,4-glucosidase with wide substrate specificity from Cytophaga hutchinsonii . Appl Microbiol Biotechnol 101, 1919–1926 (2017). https://doi.org/10.1007/s00253-016-7927-4

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