Re-engineering an in-vitro evolved ribozyme.
Ribox02 is an in-vitro evolved RNA molecule that exhibits the activity analogous to that of alcohol dehydrogenase, catalyzing redox reactions in a cofactor dependent manner. It forms a compact pseudo-knotted structure and oxidizes an alcohol seven orders of magnitude faster than the estimated spontaneous rate. To gain insights into the structure-function relationship of Ribox02, we firstly performed mutational studies to identify the critical nucleotide bases for catalysis and then engineered the ribozyme structure to improve its overall fold. The re-engineered Ribox02, named Robust, was able to catalyze the reaction approximately 25% more efficiently, leading us to generate a transacting system.
Pattnaik
Swetansu B
SB
Department of Chemistry, SUNY Buffalo, NY 14260, USA.
Suga
Hiroaki
H
D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D019020:Directed Molecular Evolution; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D016337:RNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D013329:Structure-Activity Relationship
D016337:RNA, Catalytic; D000426:Alcohol Dehydrogenase