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- Autoinducer-2 detection among commensal oral streptococci is dependent on pH and boric acid
- Giancarlo A. Cuadra , Ashley J. Frantellizzi , Kimberly M. Gaesser , Steven P. Tammariello , Anika Ahmed
- J. Microbiol. 2016;54(7):492-502. Published online June 28, 2016
- DOI: https://doi.org/10.1007/s12275-016-5507-z
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Abstract
- Autoinducer-2, considered a universal signaling molecule, is produced by many species of bacteria; including oral strains. Structurally, autoinducer-2 can exist bound to boron (borated autoinducer-2). Functionally, autoinducer-2 has been linked to important bacterial processes such as virulence and biofilm formation. In order to test production of autoinducer-2 by a given bacterial strain, a bioassay using marine bioluminescent bacteria Vibrio harveyi as a reporter for autoinducer-2 has been designed. We hypothesize that pH adjustment and addition of boron are required for optimal bioluminescence and accurate autoinducer-2 detection. Using this reporter strain we tested autoinducer-2 activity from two oral commensal species, Streptococcus gordonii DL1 and Streptococcus oralis 34. Spent broth was collected and adjusted to pH 7.5 and supplemented with boric acid prior to measuring autoinducer- 2 activity. Results show that low pH inhibits bioluminescence of the reporter strain, but pH 7.5 allows for bioluminescence induction and proper readings of autoinducer-2 activity. Addition of boric acid also has a positive effect on bioluminescence allowing for a more sensitive detection of autoinducer-2 activity. Our data suggests that although autoinducer- 2 is present in spent broth, low pH and/or low levels of boric acid become an obstacle for proper autoinducer-2 detection. For proper autoinducer-2 detection, we propose a protocol using this bioassay to include pH adjustment and boric acid addition to spent broth. Studies on autoinducer-2 activity in several bacteria species represent an important area of study as this universal signaling molecule is involved in critical bacterial phenotypes such as virulence and biofilm formation.
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Citations
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- Inhibitory effect of Lonicera japonica flos on Streptococcus mutans biofilm and mechanism exploration through metabolomic and transcriptomic analyses
Lin Wang, Ping Liu, Yulun Wu, Hairun Pei, Xueli Cao
Frontiers in Microbiology.2024;[Epub] CrossRef - New Insights into Boron Essentiality in Humans and Animals
Andrei Biţă, Ion Romulus Scorei, Tudor Adrian Bălşeanu, Maria Viorica Ciocîlteu, Cornelia Bejenaru, Antonia Radu, Ludovic Everard Bejenaru, Gabriela Rău, George Dan Mogoşanu, Johny Neamţu, Steven A. Benner
International Journal of Molecular Sciences.2022; 23(16): 9147. CrossRef - Exogenous autoinducer-2 inhibits biofilm development of Desulfovibrio sp. Huiquan2017
Ee Li, Jiajia Wu, Dun Zhang
World Journal of Microbiology and Biotechnology.2021;[Epub] CrossRef - Efficacy of Preprocedural Boric Acid Mouthrinse in Reducing Viable Bacteria in Dental Aerosols Produced during Ultrasonic Scaling
Swet Nisha, Avinash Bettahalli Shivamallu, Sheela Kumar Gujjari, Pratibha Shashikumar, Nada Musharraf Ali, Madhuri Kulkarni
Contemporary Clinical Dentistry.2021; 12(3): 282. CrossRef - D-Ribose Interferes with Quorum Sensing to Inhibit Biofilm Formation of Lactobacillus paraplantarum L-ZS9
Lei Liu, Ruiyun Wu, Jinlan Zhang, Nan Shang, Pinglan Li
Frontiers in Microbiology.2017;[Epub] CrossRef
- Inhibitory effect of Lonicera japonica flos on Streptococcus mutans biofilm and mechanism exploration through metabolomic and transcriptomic analyses
- Expression of Human Protease Inhibitor Nexin-I in Escherichia coli
- Ha, Sang Deuk , Kim, Ji Ha , Park, Hey Lyoun , Hyun, Hyung Hwan , Chung, Hyung Min , Kim Kwon, Yun Hee , Seo, Seong Yum , Ko, Jung Jae , Lee, Hyun Hwan
- J. Microbiol. 1998;36(4):283-288.
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Abstract
- Human protease inhibitor nexin-I(NX-I) cDNA(1.2Kb) was isolated from human lung cDNA library and expressed under the control of T7 promoter as a fused protein in Escherichia coli BL21 and E. coli GJ1158 by addition of IPTG and Nacl as inducers. For GJ1158, 300 mM NaCl was added for induction after the cell reached A_600=0.6. As a result, E. coli GJ1158 showed higher expression level than BL2l with lesser extent of inclusion bodies. The optimum concentration of NaCl exerting no induction effect but shortening the time to reach A_600=0.6 was 50 mM. All the results suggested that E. coli GJ1158 was a useful host for efficient expression of NX-I using NaCl as an inducer. The expressed NX-1 showed an inhibitory effect on thrombin activity. The expressed protein was purified by immobilized metal affinity column chromatography (IMAC) and characterized by digestion with enterokinase (EK).
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