원문정보
초록
영어
Engineering of cellulosic biomass-degrading enzymes are important for the purpose of cost-efficient bioenergy production. Cellulose- and hemicellulose-degrading enzymes are structurally consisted of two domains, so called a catalytic domain and a cellulose-binding domain (CBD). It has been thought that the catalytic efficiency of cellulases could be improved by protein engineering of CBDs as well as catalytic domains. Both a large size of CBD mutational library and high-throughput screening system were required for CBD engineering. First, we constructed fusion proteins composed of both alkaline phosphatase and CBD, to monitor the cellulose-binding property of CBDs. Next, microwell plates containing paper discs with different kinds of cellulosic materials were designed. Binding activities of fusion proteins could be measured quantitatively using 4-methylumbelliferyl phosphate (4-MUP) as fluorescent substrates. In this poster, we report the high-throughout fluorescence-based method for the engineering of CBDs.