Family 6 carbohydrate binding modules in beta-agarases display exquisite selectivity for the non-reducing termini of agarose chains

TY - JOUR

T1 - Family 6 carbohydrate binding modules in beta-agarases display exquisite selectivity for the non-reducing termini of agarose chains

AU - Henshaw, Joanna

AU - Horne-Bitschy, Ami

AU - van Bueren, Alicia Lammerts

AU - Money, Victoria A.

AU - Bolam, David N.

AU - Czjzek, Mirjam

AU - Ekborg, Nathan A.

AU - Weiner, Ronald M.

AU - Hutcheson, Steven W.

AU - Davies, Gideon J.

AU - Boraston, Alisdair B.

AU - Gilbert, Harry J.

PY - 2006/6/23

Y1 - 2006/6/23

N2 - Carbohydrate recognition is central to the biological and industrial exploitation of plant structural polysaccharides. These insoluble polymers are recalcitrant to microbial degradation, and enzymes that catalyze this process generally contain non-catalytic carbohydrate binding modules (CBMs) that potentiate activity by increasing substrate binding. Agarose, a repeat of the disaccharide 3,6-anhydro-alpha-L-galactose-(1,3)-beta-D-galactopyranose-(1,4), is the dominant matrix polysaccharide in marine algae, yet the role of CBMs in the hydrolysis of this important polymer has not previously been explored. Here we show that family 6 CBMs, present in two different beta-agarases, bind specifically to the non-reducing end of agarose chains, recognizing only the first repeat of the disaccharide. The crystal structure of one of these modules Aga16B-CBM6-2, in complex with neoagarohexaose, reveals the mechanism by which the protein displays exquisite specificity, targeting the equatorial O4 and the axial O3 of the anhydro-L-galactose. Targeting of the CBM6 to the non-reducing end of agarose chains may direct the appended catalytic modules to areas of the plant cell wall attacked by beta-agarases where the matrix polysaccharide is likely to be more amenable to further enzymic hydrolysis.

AB - Carbohydrate recognition is central to the biological and industrial exploitation of plant structural polysaccharides. These insoluble polymers are recalcitrant to microbial degradation, and enzymes that catalyze this process generally contain non-catalytic carbohydrate binding modules (CBMs) that potentiate activity by increasing substrate binding. Agarose, a repeat of the disaccharide 3,6-anhydro-alpha-L-galactose-(1,3)-beta-D-galactopyranose-(1,4), is the dominant matrix polysaccharide in marine algae, yet the role of CBMs in the hydrolysis of this important polymer has not previously been explored. Here we show that family 6 CBMs, present in two different beta-agarases, bind specifically to the non-reducing end of agarose chains, recognizing only the first repeat of the disaccharide. The crystal structure of one of these modules Aga16B-CBM6-2, in complex with neoagarohexaose, reveals the mechanism by which the protein displays exquisite specificity, targeting the equatorial O4 and the axial O3 of the anhydro-L-galactose. Targeting of the CBM6 to the non-reducing end of agarose chains may direct the appended catalytic modules to areas of the plant cell wall attacked by beta-agarases where the matrix polysaccharide is likely to be more amenable to further enzymic hydrolysis.

KW - RHODOTHERMUS-MARINUS XYLANASE

KW - LIGAND-BINDING

KW - NONCRYSTALLINE CELLULOSE

KW - CRYSTAL-STRUCTURES

KW - RECOGNITION

KW - DOMAIN

KW - PROTEIN

KW - SITES

KW - DEGRADATION

KW - REFINEMENT

U2 - 10.1074/jbc.M600702200

DO - 10.1074/jbc.M600702200

M3 - Article

SN - 0021-9258

VL - 281

SP - 17099

EP - 17107

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 25

ER -