Lee Dong-Heon, Oh Kye-Heon, Kahng Hyung-Yeel
Department of Environmental Education, Sunchon National University, Sunchon, 540-742, Korea.
Biotechnol Lett. 2009 Aug;31(8):1245-51. doi: 10.1007/s10529-009-0008-3. Epub 2009 Apr 29.
Exiguobacterium sp. CNU020, an alkaliphile and extremohalophile bacterium, is resistant to 20 mM H(2)O(2), a concentration that is lethal to most bacteria. Representative antioxidant genes of catalase and superoxide dismutase (SOD), obtained by PCR amplification of the genomic DNA, were characterized: the 252-bp catalase gene shared 77% similarity in the deduced amino acid sequence to that of E. oxidotolerans T-2-2(T). The 420-bp SOD gene had the closest similarity (94.3%) to the manganese-SOD of E. sibiricum 255-15. Through activity-staining analysis, stain CNU020 had at least four catalase isoforms: C1, C2, C3 and C4. Expression of each catalase isoform was dependent on the growth phase and oxidants but two catalases (C3 and C4) were always induced and expressed at a similar rate, indicating that they were constitutively expressed. RT-PCR-based expression analysis at the transcriptional level suggested that the catalase gene is strongly expressed in response to 2 mM H(2)O(2), 0.2 mM Paraquat and 0.2 mM menadione. However, the SOD gene exhibited no observable expression pattern with 2 mM H(2)O(2) despite its strong expression when exposed to Paraquat and menadione.
嗜碱嗜盐菌微小杆菌属Exiguobacterium sp. CNU020对20 mM过氧化氢具有抗性,该浓度对大多数细菌来说是致死性的。通过对基因组DNA进行PCR扩增获得了过氧化氢酶和超氧化物歧化酶(SOD)的代表性抗氧化基因,并对其进行了表征:252 bp的过氧化氢酶基因推导的氨基酸序列与耐氧化微小杆菌E. oxidotolerans T-2-2(T)的相似性为77%。420 bp的SOD基因与西伯利亚微小杆菌E. sibiricum 255-15的锰超氧化物歧化酶的相似性最高(94.3%)。通过活性染色分析,菌株CNU020至少有四种过氧化氢酶同工型:C1、C2、C3和C4。每种过氧化氢酶同工型的表达取决于生长阶段和氧化剂,但两种过氧化氢酶(C3和C4)总是被诱导并以相似的速率表达,表明它们是组成型表达的。基于RT-PCR的转录水平表达分析表明,过氧化氢酶基因在2 mM过氧化氢、0.2 mM百草枯和0.2 mM甲萘醌作用下强烈表达。然而,SOD基因在2 mM过氧化氢作用下未表现出明显的表达模式,尽管其在接触百草枯和甲萘醌时强烈表达。