KCS

Biotransformation using enzymes or whole-cell catalysts has been emerging as one of major synthetic tools in organic synthesis. As a biocatalyst, Hab mutase has been known to convert hydroxylaminobenzene to aminophenol regioselectively. Recently we have found a novel Hab mutase through High-throughput screening of metagenomics libraries, and developed a new synthetic pathway from nitrobenzenes to aminophenols. Based on this result, database search using BLASTp found 62 organisms that have similar sequence to the original HabM. Among these 62 Hab Mutase-like proteins, 30 proteins with 25-80% sequence identity were selected and phylogenetically compared using Clustal W2. Finally, 17 Hab Mutase-like proteins with least sequence similarity among each other were selected, and their gene synthesized for recombinant expression in Escherichia coli. Then we checked Hab Mutase activities of those proteins by testing the biotransformation of hydroxylaminobenzene to 2-aminophenol, and 5 novel Hab Mutases were identified.