Heterologous growth phase- and temperature-dependent expression and H2O2 toxicity protection of a superoxide-inducible monofunctional catalase gene from Xanthomonas oryzae pv. oryzae.
Catalase is an important protective enzyme against H2O2 toxicity. Here, we report the characterization of a Xanthomonas oryzae pv. oryzae catalase gene (katX). The gene was localized and its nucleotide sequence was determined. The gene codes for a 77-kDa polypeptide. The deduced katX amino acid sequence shares regions of high identity with other monofunctional catalases in a range of organisms from bacteria to eukaryotes. The transcriptional regulation of katX was atypical of bacterial monofunctional kat genes. Northern (RNA) analysis showed that katX transcription was highly induced by treatments with low concentrations of menadione, a superoxide generator, and methyl methanesulfonate, a mutagen. It was only weakly induced by H2O2. Unlike in other bacteria, a high level of catalase in Xanthomonas spp. provided protection from the growth-inhibitory and killing effects of H2O2 but not from those of organic peroxides and superoxide generators. Unexpectedly, heterologous expression of katX in Escherichia coli was both growth phase and temperature dependent. Catalase activity in E. coli kat mutants harboring katX on an expression vector was detectable only when the cells entered the stationary phase of growth and at 28 degrees C. The patterns of transcription regulation, heterologous expression, and physiological function of katX are different from previously studied bacterial kat genes.
Mongkolsuk
S
S
Laboratory of Biotechnology, Chulabhorn Research Institute, Lak Si, Bangkok, Thailand.
Loprasert
S
S
Vattanaviboon
P
P
Chanvanichayachai
C
C
Chamnongpol
S
S
Supsamran
N
N
D000595:Amino Acid Sequence; D001483:Base Sequence; D002374:Catalase / Q000235:genetics*; D003001:Cloning, Molecular; D004790:Enzyme Induction / Q000187:drug effects; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D006861:Hydrogen Peroxide / Q000633:toxicity*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013481:Superoxides / Q000494:pharmacology; D013696:Temperature; D014974:Xanthomonas / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D013481:Superoxides; D006861:Hydrogen Peroxide; D002374:Catalase