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比较分析过氧化氢酶 KatB 和 KatG 在鱼类病原菌迟缓爱德华氏菌生理适应和发病机制中的作用。

Comparative analysis of the roles of catalases KatB and KatG in the physiological fitness and pathogenesis of fish pathogen Edwardsiella tarda.

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, China.

出版信息

Lett Appl Microbiol. 2012 May;54(5):425-32. doi: 10.1111/j.1472-765X.2012.03225.x. Epub 2012 Mar 15.

Abstract

AIMS

The aim of this study was to reveal functional redundancy and variation of the two catalases KatB and KatG in Edwardsiella tarda.

METHODS AND RESULTS

Genome sequencing of fish pathogen Edw. tarda EIB202 reveals that it contains two genes putatively encoding catalases, katB (ETAE_1368) and katG (ETAE_0889). Under free-living conditions, single disruption in katB or katG resulted in no growth impairment, whereas double mutation of the two genes led to moderate decrease in growth, indicating that these two catalases were together essential for the physiological fitness by dissipating the endogenous H(2) O(2) . katG mutant exhibited much more elevated sensitivity to exogenous H(2) O(2) than katB mutant did, indicating that KatG was quasi-essential in detoxifying external reactive oxygen species (ROS) in Edw. tarda EIB202. Further comparative analysis indicated that katB or katG disruption showed different effects on the virulence-related processes of Edw. tarda such as haemolysin production, bile and serum resistance, as well as the internalization within fish epithelial cells. Moreover, both of the katB and katG mutants exhibited incapacity to replicate in murine macrophage J774 cell model, although the deficiency was seen much severe for ΔkatB/katG mutant. With regard to in vivo virulence, katB and katG mutants displayed delayed lethality and increased LD(50) values for zebrafish.

CONCLUSIONS

KatB and KatG in Edw. tarda serve for the physiological fitness, and pathogenesis related the bacterial survival in macrophage and in vivo of fish.

SIGNIFICANCE AND IMPACT OF THE STUDY

Counteracting ROS for systemic infection, Edw. tarda catalase KatG and KatB merits as potential targets for attenuated live vaccine construction.

摘要

目的

本研究旨在揭示爱德华氏菌中两种过氧化氢酶 KatB 和 KatG 的功能冗余和变化。

方法和结果

对鱼类病原体爱德华氏菌 EIB202 的基因组测序表明,它包含两个可能编码过氧化氢酶的基因,katB(ETAE_1368)和 katG(ETAE_0889)。在自由生活条件下,katB 或 katG 的单一缺失不会导致生长受损,而这两个基因的双重突变导致生长适度下降,表明这两种过氧化氢酶共同对于通过耗散内源性 H(2)O(2) 对于生理适应性是必需的。katG 突变体对外源性 H(2)O(2)的敏感性比 katB 突变体高得多,表明 KatG 在爱德华氏菌 EIB202 中对外源活性氧(ROS)的解毒是准必需的。进一步的比较分析表明,katB 或 katG 的缺失对爱德华氏菌的毒力相关过程如溶血素产生、胆汁和血清抗性以及在鱼类上皮细胞内的内化有不同的影响。此外,katB 和 katG 突变体在鼠巨噬细胞 J774 细胞模型中均无法复制,尽管 ΔkatB/katG 突变体的缺陷更为严重。就体内毒力而言,katB 和 katG 突变体在斑马鱼中表现出延迟致死性和增加的 LD(50)值。

结论

爱德华氏菌中的 KatB 和 KatG 有助于细菌在巨噬细胞和鱼类体内的生理适应性和发病机制相关的生存。

研究的意义和影响

抵御系统性感染的 ROS,爱德华氏菌过氧化氢酶 KatG 和 KatB 有潜力成为减毒活疫苗构建的潜在靶点。

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