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胶孢炭疽菌 HM3 对铜绿假单胞菌 PAO1 群体感应介导的毒力因子产生和生物膜形成的抑制作用。

Attenuation of quorum sensing mediated virulence factors production and biofilm formation in Pseudomonas aeruginosa PAO1 by Colletotrichum gloeosporioides HM3.

机构信息

Department of Microbiology, School of Life Sciences, Pondicherry University, Puducherry, 605014, India.

Fungal Biotechnology Lab, Department of Biotechnology, School of Life Sciences, Pondicherry University, Puducherry, 605014, India.

出版信息

Microb Pathog. 2021 Feb;151:104723. doi: 10.1016/j.micpath.2020.104723. Epub 2021 Jan 16.

Abstract

Signal dependent microbial communication in Pseudomonas aeruginosa PAO1 is a typical phenomenon mediated by acyl homo-serine lactone molecules that helps in developing biofilm and enhance antibiotic resistance. Microbial sources provide insight to the hidden treasure of secondary metabolites, and these structurally diversified chemical motifs can be used as antimicrobial and anti-infective agents. In the present study, endophytic fungus, Colletotrichum gloeosporioides HM3 isolated from Carica papaya leaves was explored for anti-infective potential against P. aeruginosa PAO1. The crude extract of C. gloeosporioides HM3 displayed bacteriostatic effect on P. aeruginosa PAO1 growth at 750 μg/ml concentration. A significant decline was observed in the production of quorum sensing regulated virulence factors, i.e. 56.32%, 62.54%, and 66.67% of pyocyanin, chitinase, and elastase enzyme, respectively. A drastic reduction in pathogenic determinant behaviour after treatment with crude extract of C. gloeosporioides HM3 i.e. EPS, rhamnolipid, and HCN production was noted. Light microscopy and CLSM analysis revealed that fungal extract treatment has reduced bacterial ability to form dense biofilm architecture. In silico analysis demonstrated the binding efficiency of bioactive compound, 4-(2,3-dimethoxybenzylidene)-3-methyl-1-(4-nitrophenyl)-2-pyrazolin-5-one, which is equipotent to the natural ligand and displayed a docking score of -5.436 kcal/mol with QS transcriptional regulator (LasR). Whereas the compound Acetamide, n-[tetrahydro-3-(phenylmethyl) thieno [3,4-d]thiazol-2 (3 h)-ylidene]-, s,s-dioxide exhibits a docking score of -4.088 kcal/mol (LasR) and -1.868 kcal/mol (RhlR) with cognate receptor proteins. Henceforth, the research report suggests C. gloeosporioides HM3 derived metabolites could be considered as a potential inhibitors of QS regulated virulence factors and biofilm production in P. aeruginosa PAO1.

摘要

信号依赖的微生物通讯在铜绿假单胞菌 PAO1 中是一种典型的现象,由酰基高丝氨酸内酯分子介导,有助于生物膜的形成和增强抗生素耐药性。微生物来源为次生代谢产物这一隐藏宝藏提供了深入的了解,这些结构多样化的化学基序可用作抗微生物和抗感染剂。在本研究中,从番木瓜叶中分离出的内生真菌胶孢炭疽菌 HM3 被探索用于对抗铜绿假单胞菌 PAO1 的抗感染潜力。C. gloeosporioides HM3 的粗提物在 750μg/ml 浓度下对铜绿假单胞菌 PAO1 的生长表现出抑菌作用。观察到群体感应调节的毒力因子的产量显著下降,即分别为 62.54%、66.67%的绿脓菌素、几丁质酶和弹性蛋白酶酶。用 C. gloeosporioides HM3 的粗提物处理后,致病性决定因素的行为也发生了急剧减少,即 EPS、鼠李糖脂和 HCN 的产生。荧光显微镜和 CLSM 分析显示,真菌提取物处理降低了细菌形成致密生物膜结构的能力。计算机模拟分析表明,生物活性化合物 4-(2,3-二甲氧基苄叉)-3-甲基-1-(4-硝基苯基)-2-吡唑啉-5-酮的结合效率与天然配体相当,与 QS 转录调节剂(LasR)的对接得分为-5.436kcal/mol。而化合物 Acetamide, n-[tetrahydro-3-(phenylmethyl) thieno [3,4-d]thiazol-2 (3 h)-ylidene]-, s,s-dioxide 与同源受体蛋白的对接评分分别为-4.088kcal/mol(LasR)和-1.868kcal/mol(RhlR)。因此,本研究报告表明,胶孢炭疽菌 HM3 衍生的代谢产物可被视为铜绿假单胞菌 PAO1 中 QS 调节的毒力因子和生物膜形成的潜在抑制剂。

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