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Abstract

The multi-modular non-cellulosomal endo-1,3(4)--glucanase Lic16A from contains a so-called X module (denoted as CBMX) near the N terminus of the catalytic module (191–426 aa). Melting of X-module-containing recombinant proteins revealed an independent folding of the module. CBMX was isolated and studied as a separate fragment. It was shown to bind to various insoluble polysaccharides, including xylan, pustulan, chitin, chitosan, yeast cell wall glucan, Avicel and bacterial crystalline cellulose. CBMX thus contains a hitherto unknown carbohydrate-binding module (CBM54). It did not bind soluble polysaccharides on which Lic16A is highly active. Ca ions had effects on the binding, e.g. stimulated complex formation with chitosan, which was observed only in the presence of Ca. The highest affinity to CBMX was shown for xylan (binding constant =3.1×10 M), yeast cell wall glucan (=1.4×10 M) and chitin (=3.3.10 M in the presence of Ca). Lic16A deletion derivatives lacking CBMX had lower affinity to lichenan and laminarin and a slight decrease in optimum temperature and thermostability. However, the specific activity was not significantly affected.

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2009-07-01
2020-01-21
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