Abstract
Cytophaga hutchinsonii is a gliding cellulolytic bacterium that is ubiquitously distributed in soil. The mechanism by which C. hutchinsonii achieves cellulose digestion, however, is still largely unknown. In this study, we obtained a C. hutchinsonii mutant that was defective in utilizing filter paper or Avicel as the sole carbon source by transposon mutagenesis. The interrupted gene locus, CHU_2981, encodes a hypothetical protein with only 130 amino acids. Cell fractionation and western blot detection of CHU_2981 fused with a C-terminal green fluorescence protein (GFP) indicated that CHU_2981 is located in the periplasm. The CHU_2981-disrupted mutant cells exhibited a significant growth defect on Avicel but not on glucose and cellobiose. The absence of CHU_2981 also resulted in a significant defect in colony spreading and individual cell motility compared to wild-type cells. Further analysis demonstrated that the CHU_2981-disrupted mutant cells exhibited a different profile of cellulose-absorbed outer membrane proteins from that of wild-type cells, in which protein varieties and amounts were markedly decreased. Our results showed that CHU_2981, the periplasmic non-cellulolytic protein, plays an important role in both cellulose utilization and cell motility probably by being involved in the appropriate production of outer membrane proteins.
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Acknowledgments
We are grateful for MJ McBride for providing the plasmids and strains. We thank Wei Hu for suggestions in the measurement of individual C. hutchinsonii cell motility. This work is supported by grants from the National Basic Research Program (2011CB707402) and the National Natural Science Foundation of China (31570051 and 31170762).
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Yang, T., Bu, X., Han, Q. et al. A small periplasmic protein essential for Cytophaga hutchinsonii cellulose digestion. Appl Microbiol Biotechnol 100, 1935–1944 (2016). https://doi.org/10.1007/s00253-015-7204-y
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DOI: https://doi.org/10.1007/s00253-015-7204-y