Enzymatic evidence for the presence of a critical terminal hexa-arabinoside in the cell walls of Mycobacterium tuberculosis.
A species of Cellulomonas was isolated from soil by enrichment culture and shown to secrete enzymes capable of degrading mycobacterial cell wall arabinogalactan, both the insoluble peptidoglycan-bound and base-solubilized forms. The major degradation product was purified and characterized as a hexa-arabinofuranoside, [beta-D-Araf-(1-->2)-alpha-Araf-(1-->]2-->3,5-alpha-D-Ara f-(1-->5)-D-Araf. The non-reducing ends of this unit are the sites of mycolic acid attachment and, as they also appear in lipoarabinomannan (LAM), the point of mannose capping in some mycobacteria. Thus, elaboration of the structure of this focal hexasaccharide is critical to our understanding of much of the physiology and pathogenesis of mycobacteria. The extracellular enzymes of Cellulomonas sp. also released the disaccharide, alpha-D-Araf-(1-->5)-D-Araf, from internal linear regions of arabinan and, surprisingly, convert the linear galactan backbone into cyclic oligosaccharides of the structure [-->5-D-Galf-(1-->6)-beta-D-Galf-(1-->]n where n is 2, 3 or 4. Thus, the preparation contains Schardinger-like enzyme activity. This group of enzymes are powerful tools for the dissection of the mycolylarabino-galactan-peptidoglycan (mAGP) complex of mycobacteria towards understanding its role in drug resistance, disease processes and mycobacterial physiology.
McNeil
M R
MR
Department of Microbiology, Colorado State University, Fort Collins 80523.
Robuck
K G
KG
Harter
M
M
Brennan
P J
PJ
D000192:Actinomycetales / Q000201:enzymology; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002473:Cell Wall / Q000737:chemistry; D005685:Galactans / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D013379:Substrate Specificity
D005685:Galactans; D009844:Oligosaccharides; D011135:Polysaccharides, Bacterial; D006026:Glycoside Hydrolases; C005653:arabinogalactan