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 DING Ting,LI Yong.Inhibition on quorum sensing of Pseudomonas fluorescens by cyclic dipeptide isolated from Aspergillus oryzae and its mechanism[J].Chinese Journal of Bioprocess Engineering,2020,18(02):234-244.[doi:10.3969/j.issn.1672-3678.2020.02.013]





Inhibition on quorum sensing of Pseudomonas fluorescens by cyclic dipeptide isolated from Aspergillus oryzae and its mechanism
北京大学 公共卫生学院 营养与食品卫生系,北京 100191
DING TingLI Yong
Department of Nutrition and Food Hygiene,School of Public Health,Peking University,Beijing 100191,China
群体感应 环(L-苯丙氨酸-L-脯氨酸) 荧光假单胞菌 生物被膜 分子对接
荧光假单胞菌(Pseudomonas fluorescens)是一类重要的食品腐败菌,在乳制品、肉制品、水产品等食品中经常被检测到。细菌通过群体感应(quorum sensing,QS)系统来进行彼此之间的交流,腐败菌的一些特性也受到QS系统的调控。因此,以QS系统为靶点来控制食品的腐败,提高食品质量和安全性是一条潜在可行的途径。本研究中,笔者从米曲霉(Aspergillus oryzae)D01的次级代谢产物中分离得到了一种群体感应抑制剂——环二肽(L-苯丙氨酸-L-脯氨酸,cyclo-(L-phenylalanyl-L-prolyl)),并测定了其对荧光假单胞菌群体感应的抑制活性,并用分子对接的方法分析了其抑制机制。结果表明:当环二肽的质量浓度为60 μg/mL时,细菌泳动运动、群集运动的区域直径比对照组分别减少了66.62%和36.92%; 生物被膜含量比对照组减少了75.86%。L-苯丙氨酸-L-脯氨酸能显著抑制荧光假单胞菌的QS表型——紫色素的产生、泳动和群集运动,胞外蛋白酶的产生和生物被膜的形成。分子对接分析表明,其抑制机制可能是由于环二肽与荧光假单胞菌的LuxI型蛋白竞争性结合,阻碍了信号分子产生并阻断了相关基因的表达所致。
Pseudomonas fluorescens is an important food spoilage bacterium that is frequently detected in dairy,meat and aquatic products,especially at cool temperatures.A variety of vital phenotypes are regulated by quorum sensing(QS)of spoilage bacteria.Hence,targeting the QS might be a potential and feasible approach to control the spoilage and virulence to improve food quality and safety.In this study,the quorum sensing inhibitory activity of cyclo-(L-phenylalanyl-L-prolyl),a cyclic dipeptide isolated from the metabolites of Aspergillus oryzae D01 against Pseudomonas fluorescens was determined and the inhibitory mechanism was also analyzed by molecular docking in silico.The area diameter of bacterial swimming motility,swarming motility and the content of biofilm decreased 66.62%,36.92% and 75.86%,respectively compared with the control group when the concentration of cyclic dipeptide was 60 μg/mL.The cyclo-(L-phenylalanyl-L-prolyl)significantly inhibited the QS-based phenotypes-violacein production,swimming and swarming motility,extracellular proteases production,as well as biofilm formation.Molecular docking analysis revealed that the inhibitory mechanism was probably because of the competitive binding of the cyclic dipeptide to the Lux R family transcriptional regulator of P.fluorescens,which hampered the expression of related genes.


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作者简介:丁婷(1991—),女,山东青岛人,博士,研究方向:肽营养学; 李勇(联系人),教授,E-mail:liyong@bjmu.edu.cn
DING Ting,LI Yong.Inhibition on quorum sensing of Pseudomonas fluorescens by cyclic dipeptide isolated from Aspergillus oryzae and its mechanism[J].Chin J Bioprocess Eng,2020,18(2):234-244..
更新日期/Last Update: 2020-03-31