Designing Fusion Proteins with Carbohydrate-Binding Modules Having Affinity to Enzymatically Gellable Carboxymethylcellulose Derivative Hydrogel

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  • Ashida Tomoaki
    Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University
  • Ojima Yoshihiro
    Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University
  • Sakai Shinji
    Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University
  • Sakka Makiko
    Applied Microbiology Laboratory, Graduate School of Bioresources, Mie University
  • Sakka Kazuo
    Applied Microbiology Laboratory, Graduate School of Bioresources, Mie University
  • Kawakami Koei
    Department of Chemical Engineering, Faculty of Engineering, Kyushu University
  • Taya Masahito
    Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University

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Abstract

Carboxymethylcellulose (CMC) hydrogels are promising materials for tissue engineering. In this study, we proposed a method to modify an enzymatically gellable CMC derivative (CMC-Ph) hydrogel by fusing target proteins to a carbohydrate-binding module (CBM). We designed a genetically engineered protein consisting of CBM from Clostridium thermocellum CelJ and green fluorescence protein (GFP) as a model marker. The partition coefficient of CBM-fused GFP, which is the ratio of protein concentration in the hydrogel phase to that in the liquid phase, was about 7 times higher than that of wild-type GFP. In addition, the fusion protein maintained its binding capacity towards the CMC-Ph hydrogel for about two weeks. The high binding capacity of the fusion protein was also preserved in serum-containing medium for animal cell culture. These findings demonstrated that fusing proteins on a CBM is a useful method for creating CMC-derived hydrogels.

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