Microbial Metabolism Quorum Sensing and Biofilms
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1 1 Microbial Metabolism Quorum Sensing and Biofilms Ching-Tsan Huang ( 黃慶璨 ) Office: Agronomy Hall, Room 111 Tel: (02) cthuang@ntu.edu.tw
2 2 Discovery of Cell-to-Cell Communication Stage 1 (1970s): Discovery and ecological explanation Quorum sensing Luminous bacteria (e.g. Vibrio fischeri ) contain high levels of luciferase only when cultures reach the latelog phase of growth. Autoinduction Microorganisms producing pheromones and communicating with one another, was proposed.
3 3 Discovery of Cell-to-Cell Communication Stage 2 (1980s): Proof of the Autoinduction Physiological, chemical and genetic evidences were found The structure of the autoinducer singal, N-3-(oxohexanoyl) homoserine lactone was solved. Genes required to enable autoinduction of luminescence were identified. luxr and luxi model was proved luxr: encodes an autoinducer-responsive transcriptional activator. luxi: encodes a protein required for autoinducer synthesis.
4 4 Discovery of Cell-to-Cell Communication Stage 3 (1990s): Discovery of luxr-luxi-type systems in other bacteria. Bacterium Phenotype Molecule Photobacterium fischeri Bioluminescence OHHL Vibrio harveyi Bioluminescence HBHL Erwinia carotovora Pseudomonas aeruginosa Agrobacterium tumefaciens Multiple exoenzymes virulence, antibiotic Multiple exoenzymes, virulence Virulence, secondary metabolites Ti plasmid conjugal transfer OHHL OdDHL BHL & HHL OOHL Streptomyces spp. Antibiotic, differentiation -butyrolactones & more
5 5 Quorum sensing signal molecules Acylated homoserine lactones
6 6 Mechanism of Autoinduction + More detailed
7 7 I gene and R gene LuxI-type proteins recruit substrates from general metabolism At least three LuxI-type proteins (LuxI, RhlI, TraI), the substrates are S-adenosylmethionine (AdoMet) and the appropriate fatty acid substrate conjugated to acyl carrier protein (ACP). (more) Substrate interactions of acyl-hsl synthetases The active site of an acyl-hsl synthetase must accommodate AdoMet and acyl chain carried by ACP. Regulation of acyl-hsl synthetase Regulation of enzyme activity Control of acyl-hsl synthetase
8 Pseudomonas aeruginosa lasi/rhli regulation 8
9 Quorum Sensing in Gram Positives 9
10 Bacterial QS systems 10 Appl Microbiol Biotechnol. 86(5): , 2010.
11 11 Three types of autoinducer 1 Acyl-homoserine lactone (AHL) - Autoinducer used by Gram (-) bacteria for intraspecies communication 2 Autoinducer peptide (AIP) - Autoinducer used by Gram (+) bacteria for intraspecies communication 3 Autoinducer-2 (AI-2) - Autoinducer used by many QS bacteria for interspecies communication
12 Multilingual bacteria 12 Vibrio harveyi - A bioluminescence Gram (-) marine bacterium ; free living in the ocean Communication system AI-1 molecule AI-2 molecule Speaks language unique to V. harveyi Speaks universal language used by many quorumsensing bacteria
13 Three canonical quorum-sensing circuits in bacteria 13
14 Conversation between bacteria 14 To Talk? or Not To Talk? Quorum sensing Anti-quorum sensing (Quorum quenching) Autoinducer QS Inhibitors (QSIs) Regulate QS-controlled phenotypes Natural QSIs Synthetic QSIs AHL degradation system Block QS pathways
15 15 Biofilms: Microorganisms in the real world Substratum By P. Dirckx
16 16 Characteristics of Biofilms Biologically active Biochemical reactions occur within them. Heterogeneous Discontinuous in both vertical and horizontal dimension; containing organisms, biotic and abiotic components. Dynamic Heterogeneity varies with time.
17 17 P. aeruginosa Displays Multiple Phenotypes during Development as a Biofilm Journal of Bacteriology, 184(4): , 2002.
18 18 Quorum Sensing and Biofilm Formation Biofilms are perfect situation for cell-to-cell communication High cell density may be in the region of cell per cm 3 Increased physical proximity Evidence of cell-to-cell communication within biofilms Davies et al. (1998) Science 280: & more
19 Cell/EPS distribution: wild type vs QS mutants 19 EtBr Calcofluor white WT rhli lasi
20 Engineered QS systems 20 Appl Microbiol Biotechnol. 86(5): , 2010.
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