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脲酶在肺炎克雷伯菌 CG43 酸应激反应和菌毛表达中的作用。

The role of urease in the acid stress response and fimbriae expression in Klebsiella pneumoniae CG43.

机构信息

Institute of Molecular Medicine and Bioengineering, National Yang Ming Chiao Tung University, Hsin Chu, Taiwan.

Institute of Molecular Medicine, National Tsing Hua University, Hsin Chu, Taiwan.

出版信息

J Microbiol Immunol Infect. 2022 Aug;55(4):620-633. doi: 10.1016/j.jmii.2022.02.002. Epub 2022 Mar 16.

Abstract

BACKGROUND/PURPOSE: Two urease operons were identified in Klebsiella pneumoniae CG43, ure-1 and ure-2. This study investigates whether a differential regulation of the expression of ure-1 and ure-2 exists and how urease activity influences the acid stress response and expression of type 1 and type 3 fimbriae.

METHODS

The ureA1 and ureA2 gene specific deletion mutants were constructed. Promoter activity was assessed using a LacZ reporter system. The sensitivity to acid stress was determined by assessing the survival after pH 2.5 treatment. The influence on type 1 and type 3 fimbriae expression was assessed using western blotting and mannose-sensitive yeast agglutination and biofilm formation assay, respectively.

RESULTS

Bacterial growth analysis in mM9-U or modified Stuart broth revealed that ure-1 was the principal urease system, and ure-2 had a negative effect on ure-1 activity. Deletion of the fur or nac gene had no apparent effect on the activity of P, P, and P. The P activity was enhanced by deletion of the hns gene. ureA1 deletion increased acid stress sensitivity, whereas the deleting effect of ureA2 was notable without hns. Deletion of ureA1 or ureA2 significantly induced the expression of type 1 fimbriae but decreased MrkA production and biofilm formation.

CONCLUSION

ure-1 is the primary expression system in K. pneumoniae CG43, while ure-2 is active in the absence of hns. Impairment of urease activity increases the sensitivity to acid stress, and the accumulation of urea induces the expression of type 1 fimbriae but represses type 3 fimbriae expression.

摘要

背景/目的:肺炎克雷伯氏菌 CG43 中有两个脲酶操纵子,ure-1 和 ure-2。本研究调查 ure-1 和 ure-2 的表达是否存在差异调节,以及脲酶活性如何影响酸应激反应和 1 型和 3 型菌毛的表达。

方法

构建 ureA1 和 ureA2 基因特异性缺失突变体。使用 LacZ 报告系统评估启动子活性。通过评估 pH 2.5 处理后的存活率来确定对酸应激的敏感性。使用 Western blot 法和甘露糖敏感酵母凝集试验以及生物膜形成试验分别评估对 1 型和 3 型菌毛表达的影响。

结果

在 mM9-U 或改良 Stuart 肉汤中进行细菌生长分析表明 ure-1 是主要的脲酶系统,而 ure-2 对 ure-1 活性有负面影响。fur 或 nac 基因缺失对 P、P 和 P 的活性没有明显影响。hns 基因缺失增强了 P 活性。ureA1 缺失增加了酸应激敏感性,而 ureA2 的缺失效应在没有 hns 的情况下更为显著。ureA1 或 ureA2 的缺失显著诱导了 1 型菌毛的表达,但降低了 MrkA 的产生和生物膜形成。

结论

ure-1 是肺炎克雷伯氏菌 CG43 的主要表达系统,而 ure-2 在没有 hns 的情况下活跃。脲酶活性的损害增加了对酸应激的敏感性,尿素的积累诱导了 1 型菌毛的表达,但抑制了 3 型菌毛的表达。

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