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Abstract
Bacterial cell growth and division require the coordinated action of enzymes that synthesize and degrade cell wall polymers. Here, we identify enzymes that cleave the d-arabinan core of arabinogalactan, an unusual component of the cell wall of Mycobacterium tuberculosis and other mycobacteria. We screened 14 human gut-derived Bacteroidetes for arabinogalactan-degrading activities and identified four families of glycoside hydrolases with activity against the d-arabinan or d-galactan components of arabinogalactan. Using one of these isolates with exo-d-galactofuranosidase activity, we generated enriched d-arabinan and used it to identify a strain of Dysgonomonas gadei as a d-arabinan degrader. This enabled the discovery of endo- and exo-acting enzymes that cleave d-arabinan, including members of the DUF2961 family (GH172) and a family of glycoside hydrolases (DUF4185/GH183) that display endo-d-arabinofuranase activity and are conserved in mycobacteria and other microbes. Mycobacterial genomes encode two conserved endo-d-arabinanases with different preferences for the d-arabinan-containing cell wall components arabinogalactan and lipoarabinomannan, suggesting they are important for cell wall modification and/or degradation. The discovery of these enzymes will support future studies into the structure and function of the mycobacterial cell wall.
Bacterial cell growth and division require the coordinated action of enzymes that synthesize and degrade cell wall polymers. Here, the authors identify enzymes that cleave the D-arabinan core of arabinogalactan, an unusual component of the cell wall of Mycobacterium tuberculosis and other mycobacteria.
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1 Newcastle University, Newcastle University Biosciences Institute, Medical School, Newcastle upon Tyne, UK (GRID:grid.1006.7) (ISNI:0000 0001 0462 7212)
2 University of Birmingham, Institute of Microbiology and Infection, School of Biosciences, Birmingham, UK (GRID:grid.6572.6) (ISNI:0000 0004 1936 7486)
3 University of Melbourne, Parkville, School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, Victoria, Australia (GRID:grid.1008.9) (ISNI:0000 0001 2179 088X)
4 Massachusetts Institute of Technology, Department of Chemistry, Cambridge, USA (GRID:grid.116068.8) (ISNI:0000 0001 2341 2786); Massachusetts Institute of Technology, The Broad Institute of Harvard and MIT, Cambridge, MA, USA The Koch Integrative Cancer Research Institute, Cambridge, USA (GRID:grid.116068.8) (ISNI:0000 0001 2341 2786)
5 University of Liverpool, Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, Liverpool, UK (GRID:grid.10025.36) (ISNI:0000 0004 1936 8470)
6 University of Essex, School of Life Sciences, Colchester, UK (GRID:grid.8356.8) (ISNI:0000 0001 0942 6946)
7 King Abdulaziz University, Department of Biological Sciences, Jeddah, Saudi Arabia (GRID:grid.412125.1) (ISNI:0000 0001 0619 1117); Technical University of Denmark, Department of Biotechnology and Biomedicine (DTU Bioengineering), Lyngby, Denmark (GRID:grid.5170.3) (ISNI:0000 0001 2181 8870)
8 Northumbria University, Microbial Enzymology Group, Department of Applied Sciences, Newcastle upon Tyne, UK (GRID:grid.42629.3b) (ISNI:0000000121965555)
9 Massachusetts Institute of Technology, Department of Chemistry, Cambridge, USA (GRID:grid.116068.8) (ISNI:0000 0001 2341 2786)