X4 modules represent a new family of carbohydrate-binding modules that display novel properties.
The hydrolysis of the plant cell wall by microbial glycoside hydrolases and esterases is the primary mechanism by which stored organic carbon is utilized in the biosphere, and thus these enzymes are of considerable biological and industrial importance. Plant cell wall-degrading enzymes in general display a modular architecture comprising catalytic and non-catalytic modules. The X4 modules in glycoside hydrolases represent a large family of non-catalytic modules whose function is unknown. Here we show that the X4 modules from a Cellvibrio japonicus mannanase (Man5C) and arabinofuranosidase (Abf62A) bind to polysaccharides, and thus these proteins comprise a new family of carbohydrate-binding modules (CBMs), designated CBM35. The Man5C-CBM35 binds to galactomannan, insoluble amorphous mannan, glucomannan, and manno-oligosaccharides but does not interact with crystalline mannan, cellulose, cello-oligosaccharides, or other polysaccharides derived from the plant cell wall. Man5C-CBM35 also potentiates mannanase activity against insoluble amorphous mannan. Abf62A-CBM35 interacts with unsubstituted oat-spelt xylan but not substituted forms of the hemicellulose or xylo-oligosaccharides, and requires calcium for binding. This is in sharp contrast to other xylan-binding CBMs, which interact in a calcium-independent manner with both xylo-oligosaccharides and decorated xylans.
School of Cell and Molecular Biosciences, University of Newcastle upon Tyne, The Agriculture Building, Newcastle upon Tyne NE1 7RU, United Kingdom.
David N
DN
Bolam
Hefang
H
Xie
Gavin
G
Pell
Deborah
D
Hogg
Greta
G
Galbraith
Bernard
B
Henrissat
Harry J
HJ
Gilbert
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D050260:Carbohydrate Metabolism; D020633:Cellvibrio; D006026:Glycoside Hydrolases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010944:Plants / Q000378:metabolism / Q000382:microbiology; D011485:Protein Binding; D016415:Sequence Alignment
D006026:Glycoside Hydrolases