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重组 PvdQ 工程菌对生物膜形成的群体感应淬灭潜力。

Quorum-quenching potential of recombinant PvdQ-engineered bacteria for biofilm formation.

机构信息

College of Fisheries and Life Science, Shanghai Ocean University, Nanhui New City, No.999, Huchenghuan Rd, Shanghai, People's Republic of China.

Key Laboratory of Tropical and Subtropical Fishery Resource Application and Cultivation, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences (CAFS), No.1 Xingyu Road, Xilang, Guangzhou, Liwan District, 510380, People's Republic of China.

出版信息

Int Microbiol. 2023 Aug;26(3):639-650. doi: 10.1007/s10123-023-00329-1. Epub 2023 Feb 11.

Abstract

Quorum sensing (QS) is a core mechanism for bacteria to regulate biofilm formation, and therefore, QS inhibition or quorum quenching (QQ) is used as an effective and economically feasible strategy against biofilms. In this study, the PvdQ gene encoding AHL acylase was introduced into Escherichia coli (DE3), and a PvdQ-engineered bacterium with highly efficient QQ activity was obtained and used to inhibit biofilm formation. Gene sequencing and western blot analysis showed that the recombinant pET-PvdQ strain was successfully constructed. The color reaction of Agrobacterium tumefaciens A136 indicated that PvdQ engineering bacteria had shown strong AHL signal molecule quenching activity and significantly inhibited the adhesion (motility) of Pseudomonas aeruginosa and biofilm formation of activated sludge bacteria in Membrane Bio-Reactor (MBR; inhibition rate 51-85%, p < 0.05). In addition, qRT-PCR testing revealed that recombinant PvdQ acylase significantly reduced the transcription level of QS biofilm formation-related genes (cdrA, pqsA, and lasR; p < 0.05). In this study, QQ genetically engineered bacteria enhanced by genetic engineering could effectively inhibit the QS signal transduction mechanism and have the potential to control biofilm formation of pathogenic bacteria in the aquaculture environment, providing an environmentally friendly and alternative antibiotic strategy to suppress biofilm contamination.

摘要

群体感应 (QS) 是细菌调节生物膜形成的核心机制,因此,QS 抑制或群体淬灭 (QQ) 被用作对抗生物膜的有效且经济可行的策略。在本研究中,引入了编码 AHL 酶的 PvdQ 基因到大肠杆菌 (DE3) 中,获得了具有高效 QQ 活性的 PvdQ 工程菌,并用于抑制生物膜形成。基因测序和 Western blot 分析表明,成功构建了重组 pET-PvdQ 菌株。农杆菌 A136 的显色反应表明,PvdQ 工程菌具有很强的 AHL 信号分子淬灭活性,并显著抑制了铜绿假单胞菌的粘附(运动)和膜生物反应器(MBR)中活性污泥细菌的生物膜形成(抑制率为 51-85%,p < 0.05)。此外,qRT-PCR 测试表明,重组 PvdQ 脂肪酶显著降低了 QS 生物膜形成相关基因 (cdrA、pqsA 和 lasR) 的转录水平(p < 0.05)。在本研究中,通过基因工程增强的 QQ 遗传工程菌可以有效地抑制 QS 信号转导机制,并有可能控制水产养殖环境中病原菌的生物膜形成,为抑制生物膜污染提供了一种环保和替代抗生素的策略。

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