Mongkolsuk S, Loprasert S, Vattanaviboon P, Chanvanichayachai C, Chamnongpol S, Supsamran N
Laboratory of Biotechnology, Chulabhorn Research Institute, Lak Si, Bangkok, Thailand.
J Bacteriol. 1996 Jun;178(12):3578-84. doi: 10.1128/jb.178.12.3578-3584.1996.
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.
过氧化氢酶是一种抵御过氧化氢毒性的重要保护酶。在此,我们报道了水稻白叶枯病菌过氧化氢酶基因(katX)的特性。该基因已定位并测定了其核苷酸序列。该基因编码一种77 kDa的多肽。推导的katX氨基酸序列与从细菌到真核生物等一系列生物体中的其他单功能过氧化氢酶具有高度同源区域。katX的转录调控不同于细菌单功能kat基因。Northern(RNA)分析表明,katX转录受到低浓度甲萘醌(一种超氧化物产生剂)和甲磺酸甲酯(一种诱变剂)处理的高度诱导。它仅受到过氧化氢的微弱诱导。与其他细菌不同,黄单胞菌属中的高水平过氧化氢酶可保护其免受过氧化氢的生长抑制和杀伤作用,但不能免受有机过氧化物和超氧化物产生剂的影响。出乎意料的是,katX在大肠杆菌中的异源表达既依赖生长阶段又依赖温度。在表达载体上携带katX的大肠杆菌kat突变体中的过氧化氢酶活性仅在细胞进入生长稳定期且温度为28摄氏度时才可检测到。katX的转录调控模式、异源表达模式和生理功能与先前研究的细菌kat基因不同。