In vitro synthesis of AI-2 and its effect on tetracycline resistance of Vibrio parahaemolyticus

LIN Cai-Yun1,2 JIANG Yan-Hua2 YAO Lin2 LI Feng-Ling2 QU Meng2 WANG Lian-Zhu2 XU Jia-Chao1

(1.College of Food Science and Engineering, Ocean University of China, Qingdao, Shandong, China 266003)
(2.Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, Shandong, China 266071)

【Abstract】[Background] Antimicrobial resistance of bacteria has become more and more serious due to the abuse of antimicrobials. As a major foodborne pathogen, Vibrio parahaemolytics also exhibits a certain level of antimicrobial resitance. Quorum sensing system can regulate the antimicrobial resistance of bacteria, which provides a new pathway to study the mechanism and control technique of antimicrobial resistance of V. parahaemolytics. [Objective] To study the effect of signaling molecule autoinducer-2 (AI-2) on tetracycline resistance of V. parahaemolyticus strains from seafood. [Methods] AI-2 was synthesized in vitro by a reaction using the critical enzymes, S-ribosylhomocysteinase (LuxS) and S-adenosylhomocysteine nucleosidase (Pfs), which were prepared by prokaryotic expression. The effect of AI-2 on tetracycline resistance of V. parahaemolyticus was determined by colony counting method. The effect of AI-2 at different concentrations on the transcriptional levels of tetracycline resistance genes in V. parahaemolyticus was assayed by reverse transcription and real-time quantitative PCR. [Results] LuxS and Pfs enzymes were obtained by prokaryotic expression. The bioactive AI-2 could be synthesized in vitro by adding LuxS and Pfs to the substrate S-adenosylhomocysteine (SAH), with the fluorescence intensity of about 6 times as much as that of positive control. When treated with tetracycline at subinhibitory concentration, AI-2 could significantly promote the growth of V. parahaemolyticus strains, and AI-2 at the concentrations of 6, 15 and 30 μmol/L could increase the transcriptional levels of tetracycline resistance genes in V. parahaemolyticus strains to a certain extent. [Conclusion] AI-2 could enhance the tetracycline resistance of V. parahaemolyticus, which provided a reference for further study on the antimicrobial resistance mechanism of V. parahaemolyticus and developing control techniques targeting AI-2 on the antimicrobial resistance of V. parahaemolyticus.

【Keywords】 Vibrio parahaemolyticus; Signaling molecule AI-2; S-ribosylhomocysteinase; S-adenosylhomocysteine nucleosidase; Tetracycline resistance; Tetracycline resistance gene tet;

【DOI】

【Funds】 National Natural Science Foundation of China (31601566) Central Public-interest Scientific Institution Basal Research Fund of Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences (20603022020004) National Key Research and Development Program of China (2017YFC1600703)

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(Translated by QI RS)

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This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 47, No. 05, Pages 1321-1331

May 2020

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results
  • 3 Discussion
  • 4 Conclusions
  • References