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Co‐operative binding of triplicate carbohydrate‐binding modules from a thermophilic xylanase
Author(s) -
Boraston Alisdair B.,
McLean Bradley W.,
Chen Grace,
Li Anson,
Warren R. Antony J.,
Kilburn Douglas G.
Publication year - 2002
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.2002.02730.x
Subject(s) - carbohydrate binding module , avidity , thermophile , biology , affinities , dna , xylan , polysaccharide , biochemistry , strain (injury) , carbohydrate , glycan , cellulose , crystallography , stereochemistry , chemistry , genetics , enzyme , anatomy , antibody , cellulase , glycoprotein
Summary Family 6 carbohydrate‐binding modules were am‐plified by polymerase chain reaction (PCR) from Clostridium stercorarium strain NCIB11754 genomic DNA as a triplet. Individually, these modules bound to xylooligosaccharides and cellooligosaccharides with affinities varying from ≈ 3 × 10 3 M –1 to ≈ 1 × 10 5 M –1 . Tandem and triplet combinations of these modules bound co‐operatively to soluble xylan and insoluble cellulose to give ≈ 20‐ to ≈ 40‐fold increases in affinity relative to the individual modules. This co‐operativity was an avidity effect resulting from the modules within the tandems and triplet interacting simul‐ taneously with proximal binding sites on the polysac‐charides. This occurred by both intrachain and interchain interactions. The duplication or triplication of modules appears to be linked to the growth temperature of the organism; co‐operativity in these multiplets may compensate for the loss of affinity at higher temperatures.

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