Rational design of an artificial hydrolytic nuclease by introduction of a sodium copper chlorophyllin in L29E myoglobin

Heme proteins have recently emerged as promising artificial metalloenzymes for catalyzing diverse reactions. In this report, L29E Mb, a single mutant of myoglobin (Mb), was reconstituted by replacing the heme with a sodium copper cholorophyllin (CuCP) to form a new green artificial enzyme (named CuCP-L29E Mb). The reconstituted protein CuCP-L29E Mb was found to exhibit hydrolytic DNA cleavage activity, which was not depending on O2. In addition, Mg2+ ion could effectively promote the DNA cleavage activity of CuCP-L29E Mb. Wild-type (WT) Mb reconstituted with CuCP (named CuCP-WT Mb) did not show DNA cleavage activity under the same conditions. This study suggests that both Mg2+ and the ligand Glu29 are critical for the nuclease activity and the artificial nuclease of Mg2+-CuCP-L29E Mb may have potential applications in the future.