Supplementary Information
Non-antibiotic quorum sensing inhibitorsacting against N-acyl homoserine
lactone synthase as druggable target
Chien-Yi Chang1,2, Thiba Krishnan3, Hao Wang4, Ye Chen5, Wai-Fong Yin3, Yee-Meng Chong3, Li Ying Tan3, Teik Min Chong3, Kok-Gan Chan3*
1 Interdisciplinary Computing and Complex BioSystems (ICOS) research group,School of Computing Science,Claremont Tower, Newcastle University, NewcastleuponTyneNE17RU, UK
2 The Centre for Bacterial Cell Biology, Medical School, Newcastle University,Richardson Road, Newcastle upon Tyne, NE2 4AX, UK
3Division of Genetics and Molecular Biology, Institute of Biological Sciences,
Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia
4 School of Pharmacy, Ningxia Medical University, Yinchuan, P. R. China
5 School of Life Sciences, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK
*Correspondence: , Tel. +603-79675162; Fax: +603-79674509
Supplementary Fig. S1Construction of LasI (A) and RhlI (B)L-arabinose induction plasmids. lasI and rhlI open reading frames (ORFs) with introduced XbaI and HindIII sites were amplified and cloned into the same restriction enzymes digested pBAD24. Two ORFs were driven by PBAD promoter with L-arabinose induction and terminated by rrnB terminators. pBR322 ori is a low copy number replicon in E. coli.
Supplementary Fig. S2L-arabinose-dependent induction of AHLs production detected by E. coli JM109 [pSB1075] (A) and E. coli JM109 [pSB536] (B). E. coli MG1655 carried pBAD-lasI and pBAD-rhlI were induced with 0%, 0.1% and 1% L-arabinose respectively for 4 hours. E. coli MG1655 without plasmid and with empty pBAD24 were used as negative controls. P. aeruginosa and 50 pmol synthetic C4-HSL and 3-oxo-C12-HSL were spotted as positive control. LCMS analysis of extractions of P. aeruginosa overnight culture and 4-hours induced E. coli MG1655 expressing LasI and RhlI showed the specific AHL production form each construction (C).
Supplementary Fig. S3Luminescent output measurement from E. coli MG1655 [pBAD-lux] treated with of 0.06 mM tannic acid and 2.27 mM trans-cinnamaldehyde (A). Growth curves of E. coli MG1655 [pBAD-lux] (B) and P. aeruginosa(C) in presence of 0.06 mM tannic acid and 2.27 mM trans-cinnamaldehyde.
Supplementary Table S1 Summary of LCMS result of detecting C4-HSL
Treatment / m/z value / Retention time (minute)Synthetic AHL standard / 172.3 / 0.588
Untreated negative control / 172.2 / 0.588
DMSO solvent control / 172.3 / 0.425
Salicylic acid / 172.3 / 0.556
Tannic acid / −a / −
Trans-cinnamaldehyde / − / −
aundetectable
Supplementary Table S2Summary of LCMS result of detecting 3-oxo-C12-HSL
Treatment / m/z value / Retention time (minute)Synthetic AHL standard / 298.3 / 5.785
Untreated negative control / 298.3 / 5.851
DMSO solvent control / 298.3 / 5.851
Salicylic acid / 298.3 / 5.720
Tannic acid / 298.3 / 5.883
Trans-cinnamaldehyde / 298.3 / 5.883
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