Reconstruction of Mycobacterial Dehalogenase Rv2579 by Cumulative Mutagenesis of Haloalkane Dehalogenase LinB
Nagata, Y., Prokop, Z., Marvanova, S., Sykorova, J., Monincova, M., Tsuda, M., Damborsky, J.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY 69: 2349-2355 (2003)
The homology model of protein Rv2579 from Mycobacterium tuberculosis H37Rv was compared with the crystal structure of haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26 and uncovered that six out of nineteen amino acid residues which form an active site and entrance tunnel are different between LinB and Rv2579. To characterize an effect of replacements of these six amino acid residues, mutations were introduced cumulatively into the six amino acid residues of LinB. The six-fold mutant which is supposed to have an active site of Rv2579 showed haloalkane dehalogenase activity with tested haloalkanes, confirming that Rv2579 is a member of haloalkane dehalogenase protein family.