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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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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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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DOI: https://doi.org/10.1007/s00253-016-7927-4