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Key amino acid residues for the endo-processive activity of GH74 xyloglucanase
Tomohiko Matsuzawa1, Yuji Saito2, Katsuro Yaoi1
1Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tukuba Central 6, 1-1-1 Higashi, Tukuba, Ibaraki 305-8566, Japan.
Abstract:
Unlike endo-dissociative-xyloglucanases, Paenibacillus XEG74 is an endo-processive xyloglucanase that contains four unique tryptophan residues in the negative subsites (W61 and W64) and the positive subsites (W318 and W319), as indicated by three-dimensional homology modelling. Selective replacement of the positive subsite residues with alanine mutations reduced the degree of processive activity and resulted in the more endo-dissociative-activity. The results showed that W318 and W319, which are found in the positive subsites, are essential for processive degradation and are responsible for maintaining binding interactions with xyloglucan polysaccharide through a stacking effect.
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