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从帕尔卡湾沉积物中分离出的产AHL-内酯酶菌株可减轻革兰氏阴性细菌病原体中群体感应介导的毒力产生 。

AHL-Lactonase Producing sp. From Palk Bay Sediment Mitigates Quorum Sensing-Mediated Virulence Production in Gram Negative Bacterial Pathogens.

作者信息

Packiavathy Issac Abraham Sybiya Vasantha, Kannappan Arunachalam, Thiyagarajan Sivaprakasam, Srinivasan Ramanathan, Jeyapragash Danaraj, Paul John Bosco John, Velmurugan Pazhanivel, Ravi Arumugam Veera

机构信息

Department of Biotechnology, Karpagam Academy of Higher Education, Coimbatore, India.

Department of Biotechnology, Alagappa University, Karaikudi, India.

出版信息

Front Microbiol. 2021 Apr 14;12:634593. doi: 10.3389/fmicb.2021.634593. eCollection 2021.

Abstract

Quorum sensing (QS) is a signaling mechanism governed by bacteria used to converse at inter- and intra-species levels through small self-produced chemicals called N-acylhomoserine lactones (AHLs). Through QS, bacteria regulate and organize the virulence factors' production, including biofilm formation. AHLs can be degraded by an action called quorum quenching (QQ) and hence QQ strategy can effectively be employed to combat biofilm-associated bacterial pathogenesis. The present study aimed to identify novel bacterial species with QQ potential. Screening of Palk Bay marine sediment bacteria for QQ activity ended up with the identification of marine bacterial isolate 28 (MSB-28), which exhibited a profound QQ activity against QS biomarker strain ATCC 12472. The isolate MSB-28 was identified as sp. through 16S-rRNA sequencing. sp. also demonstrated a pronounced activity in controlling the biofilm formation in different bacteria and biofilm-associated virulence factors' production in PAO1. Solvent extraction, heat inactivation, and proteinase K treatment assays clearly evidence the enzymatic nature of the bioactive lead. Furthermore, AHL's lactone ring cleavage was confirmed with experiments including ring closure assay and chromatographic analysis, and thus the AHL-lactonase enzyme production in sp. To conclude, this is the first report stating the AHL-lactonase mediated QQ activity from marine sediment bacteria sp. Future work deals with the characterization, purification, and mass cultivation of the purified protein and should pave the way to assessing the feasibility of the identified protein in controlling QS and biofilm-mediated multidrug resistant bacterial infections in mono or multi-species conditions.

摘要

群体感应(QS)是一种由细菌控制的信号传导机制,用于通过自身产生的称为N-酰基高丝氨酸内酯(AHLs)的小分子在种间和种内水平进行交流。通过群体感应,细菌调节和组织毒力因子的产生,包括生物膜形成。AHLs可以通过一种称为群体猝灭(QQ)的作用被降解,因此QQ策略可以有效地用于对抗与生物膜相关的细菌发病机制。本研究旨在鉴定具有QQ潜力的新型细菌物种。对帕尔卡湾海洋沉积物细菌进行QQ活性筛选,最终鉴定出海洋细菌分离株28(MSB-28),它对QS生物标志物菌株ATCC 12472表现出显著的QQ活性。通过16S-rRNA测序将分离株MSB-28鉴定为 sp. sp. 在控制不同细菌中的生物膜形成以及PAO1中与生物膜相关的毒力因子产生方面也表现出显著活性。溶剂萃取、热失活和蛋白酶K处理试验清楚地证明了生物活性先导物的酶性质。此外,通过包括闭环试验和色谱分析在内的实验证实了AHL的内酯环裂解,从而确定了 sp. 中AHL-内酯酶的产生。总之,这是第一份报道海洋沉积物细菌 sp. 中AHL-内酯酶介导的QQ活性的报告。未来的工作将涉及纯化蛋白的表征、纯化和大规模培养,应为评估所鉴定蛋白在单种或多种条件下控制QS和生物膜介导的多重耐药细菌感染的可行性铺平道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05e4/8079732/a5a437c709c5/fmicb-12-634593-g001.jpg

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