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A CBM20 low-affinity starch-binding domain from glucan, water dikinase |
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| Authors: | Camilla Christiansen Maher Abou Hachem Anders Viksø-Nielsen Birte Svensson Andreas Blennow |
| Institution: | a VKR Research Centre, Pro-Active Plants, Department of Plant Biology and Biotechnology, Faculty of Life Sciences, University of Copenhagen, DK-1871 Frederiksberg, Denmark b Enzyme and Protein Chemistry, Department of Systems Biology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark c Novozymes A/S, DK-2880 Bagsvaerd, Denmark d Department of Biology, University of Copenhagen, DK-2100 Copenhagen Ø, Denmark |
| Abstract: | The family 20 carbohydrate-binding module (CBM20) of the Arabidopsis starch phosphorylator glucan, water dikinase 3 (GWD3) was heterologously produced and its properties were compared to the CBM20 from a fungal glucoamylase (GA). The GWD3 CBM20 has 50-fold lower affinity for cyclodextrins than that from GA. Homology modelling identified possible structural elements responsible for this weak binding of the intracellular CBM20. Differential binding of fluorescein-labelled GWD3 and GA modules to starch granules in vitro was demonstrated by confocal laser scanning microscopy and yellow fluorescent protein-tagged GWD3 CBM20 expressed in tobacco confirmed binding to starch granules in planta. |
| Keywords: | CBM carbohydrate-binding module CLSM confocal laser scanning microscopy GA glucoamylase GWD glucan water dikinase SBD starch-binding domain SPR surface plasmon resonance YFP yellow fluorescent protein |
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