Search for a dextransucrase minimal motif involved in dextran binding |
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Authors: | Suwannarangsee Surisa Moulis Claire Potocki-Veronese Gabrielle Monsan Pierre Remaud-Simeon Magali Chulalaksananukul Warawut |
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Affiliation: | Biological Science Program, Faculty of Science, Chulalongkorn University, 254 Phyathai Road, Patumwan, Bangkok, Thailand. |
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Abstract: | Fourteen truncated forms of Leuconostoc mesenteroides NRRL B512-F dextransucrase, involving N-, C- or N- plus C-terminal domain truncations were tested for their ability to bind dextrans. The shortest fragment (14kDa molecular weight) that still exhibited a strong interaction with dextran was localized between amino acids N1397 and A1527 of the C-terminal domain (GBD-7) and consists of six YG repeats. With a dissociation constant K(d) of 2.8x10(-9)M, this motif shows a very high affinity for isomaltohexaose and longer dextrans, supporting the proposed role of GBD in polymer formation. The potential application of GBD-7 as an affinity tag onto cheap resins like Sephacryl S300HR for rapid purification was evaluated and is discussed. |
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Keywords: | DSR-S, Leuconostoc mesenteroides NRRL B512-F dextransucrase GBD, glucan-binding domain GH, glycoside-hydrolase His, polyhistidine tag |
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