Thiocarbamate herbicide-inducible nonheme haloperoxidase of Rhodococcus erythropolis NI86/21.
During biodegradation of thiocarbamate herbicides by Rhodococcus erythropolis NI86/21, a protein with an M(r) of 30,000 is induced (I. Nagy, G. Schoofs, F. Compernolle, P. Proost, J. Vanderleyden, and R.De Mot, J. Bacteriol. 177:676-687, 1995). Based on N-terminal sequence data for the protein purified by two-dimensional electrophoresis, the corresponding structural gene, thcF, was cloned and sequenced. The deduced protein sequence of ThcF is homologous to those of nonheme haloperoxidases. A particularly high level of sequence identity (72.6%) was observed for the chloroperoxidase from Pseudomonas pyrrocinia. A polyclonal antibody against the latter enzyme cross-reacted with ThcF either produced by the original Rhodococcus cells or overexpressed heterologously in Escherichia coli. In both thiocarbamate-grown Rhodococcus cells and E. coli cells expressing thcF, the haloperoxidase activity of ThcF was demonstrated. The thiocarbamate-inducible R. erythropolis ThcF protein represents the first (nonheme) haloperoxidase to be identified in a nocardioform actinomycete.
De Schrijver
A
A
F.A. Janssens Laboratory of Genetics, Catholic University of Leuven, Heverlee, Belgium.
Nagy
I
I
Schoofs
G
G
Proost
P
P
Vanderleyden
J
J
van Pée
K H
KH
De Mot
R
R
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology; D001483:Base Sequence; D001673:Biodegradation, Environmental; D015139:Blotting, Southern; D015153:Blotting, Western; D002711:Chloride Peroxidase / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D003429:Cross Reactions / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D004790:Enzyme Induction / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D010544:Peroxidases / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D011549:Pseudomonas / Q000235:genetics; D012240:Rhodococcus / Q000201:enzymology* / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013859:Thiocarbamates / Q000378:metabolism
D000907:Antibodies, Bacterial; D013859:Thiocarbamates; D010544:Peroxidases; D002711:Chloride Peroxidase