Salt-Switchable Artificial Cellulase Regulated by a DNA Aptamer.
A novel artificial cellulase was developed by conjugating a DNA aptamer to an endoglucanase catalytic domain, thereby substituting the natural carbohydrate-binding module. Circular dichroism spectroscopy and adsorption isotherm showed the binding motif of cellulose-binding DNA aptamer (CelApt) was G-quadruplex and stem-loop structures stabilized in the presence of salts, and CelApt binding preferred the amorphous region of the solid cellulose. By introducing the revealed salt-switchable cellulose-binding nature of CelApt into a catalytic domain of a cellulase, we created CelApt-catalytic domain conjugate possessing both controllable adsorption on the solid substrates and equal enzymatic activity to the wild-type cellulase. Thus potential use of a responsive DNA aptamer for biocatalysis at a solid surface was demonstrated.
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University , 744 Motooka, Fukuoka 819-0395, Japan.
Mari
M
Takahara
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University , 744 Motooka, Fukuoka 819-0395, Japan.
Geisa Aparecida Lopes Gonçalves
GA
Budinova
Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University , 6-6-116 Aoba, Aramaki, Aoba-ku, Sendai 980-8579, Japan.
Hikaru
H
Nakazawa
Biomass Engineering Program, RIKEN , 1-7-22, Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
Yutaro
Y
Mori
Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University , 6-6-116 Aoba, Aramaki, Aoba-ku, Sendai 980-8579, Japan.
Mitsuo
M
Umetsu
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University , 744 Motooka, Fukuoka 819-0395, Japan.|Division of Biotechnology, Center for Future Chemistry, Kyushu University , 744 Motooka, Fukuoka 819-0395, Japan.
Noriho
N
Kamiya
D000327:Adsorption; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D055162:Biocatalysis; D020134:Catalytic Domain / Q000187:drug effects; D002480:Cellulase / Q000737:chemistry*; D002482:Cellulose / Q000031:analogs & derivatives* / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D054856:G-Quadruplexes / Q000187:drug effects; D012965:Sodium Chloride / Q000494:pharmacology
D052157:Aptamers, Nucleotide; C033190:DNA-cellulose; D012965:Sodium Chloride; D002482:Cellulose; D004247:DNA; D002480:Cellulase