Reconstruction of mycobacterial dehalogenase Rv2579 by cumulative mutagenesis of haloalkane dehalogenase LinB.
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 this analysis revealed that 6 of 19 amino acid residues which form an active site and entrance tunnel are different in LinB and Rv2579. To characterize the effect of replacement of these six amino acid residues, mutations were introduced cumulatively into the six amino acid residues of LinB. The sixfold mutant, which was supposed to have the active site of Rv2579, exhibited haloalkane dehalogenase activity with the haloalkanes tested, confirming that Rv2579 is a member of the haloalkane dehalogenase protein family.
Nagata
Yuji
Y
Department of Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai 980-8577, Japan. aynaga@ige.tohoku.ac.jp
Prokop
Zbynek
Z
Marvanová
Sona
S
Sýkorová
Jana
J
Monincová
Marta
M
Tsuda
Masataka
M
Damborský
Jirí
J
D000473:Alkanes / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D006867:Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics; D016415:Sequence Alignment; D020578:Sphingomonas / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
D000473:Alkanes; D006867:Hydrolases; C046137:haloalkane dehalogenase