Fig. 4

In silico model of AzoC and reconstitution of nucleophilic activity of AzoC via sequence shuffling. a Structure model of AzoC. The iron atoms are shown as brown spheres. The two iron ions are coordinated with protein residues Glu-102, Glu-137, His-140, Glu-198, His- 225, Glu-229, and His-232.The mutated sequence of AzoC-mutant is shown as a blue ribbon. b HPLC analysis of AzoC-P and AzoC-mutant-P’s in vitro biochemical reactions at 345 nm. Reactions were performed in 20 mM HEPES buffer (pH 7). (i) Standard 8, (ii) standard 10, (iii) 8 with eq AzoC-P for 1 min, (iv) 8 with eq AzoC-mutant-P for 1 min