Quorum sensing and applications in biotechnology

Quorum sensing and
applications in biotechnology
By: Roghayeh Shirvani
Supervisor: Dr Mohammad Barshan
May.2017
University of Tehran
Department of Life Science
Engineering
2

3
History of quorum
sensing
Quorum sensing
in Prokaryotes
Quorum sensing
in Eukaryotes
Applications of
QS in medical
biotechnology
Applications of
QS in industrial
biotechnology
Applications of
QS in agricultural
biotechnology

TheStory of
Quorum
sensing
Hastings et al,1974:
The first evidence for cell to cell
communication in vibrio spp.
Greenberg et al, 1996 :
Cell to cell communication in bacteria
= Quorum sensing.
Bonnie Bassler et al, 1999 :
Molecular mechanism of quorum
sensing.
Lee, Anna Jisu, and LingchongYou. "Cells Listen toTheir InnerVoice." Science 343.6171 (2014). 4

Hawaiian
bobtail squid
(Euprymna
scolopes)
• Quorum sensing was
first discovered in the
marine luminescent
bacterium Vibrio fisheri
symbiont of the squid
Euprymna scolopes.
Miller, Melissa B., and Bonnie L. Bassler. "Quorum sensing in bacteria." Annual Reviews in Microbiology 55.1 (2001) 5

One forAll
andAll for
One !
The word “Quorum” in this
sense means: sufficient
numbers.
Example of quorum
sensing controlled
processes:
bioluminescence, biofilm
formation, virulence factor
expression, antibiotic
production, sporulation,
and competence for DNA
uptake.
“Sociomicrobiology”
Gobbetti, M., et al. "Cell–cell communication in food related bacteria." International journal of food microbiology 120.1 (2007). 6

7
History of quorum
sensing
Quorum sensing
in Prokaryotes
Quorum sensing
in Eukaryotes
Applications of
QS in medical
biotechnology
Applications of
QS in industrial
biotechnology
Applications of
QS in agricultural
biotechnology

Quorum sensingmoleculesin Bacteria
I. Lux mediated system (AHL- mediatedQS)
II. Peptide mediated system (AIP- mediatedQS)
III. AI-2 system
Taj, MUHAMMAD KAMRAN, et al. "Quorum sensing and its different signals systems in bacteria." Impact International Journal of Research
in Applied Natural & Social Sciences 2 (2014)
8

AI-1System
Gram - negative
bacteria
 Interspecies
communication.
 Acylated hemoserine
lactones.(AHLs)
 LuxI and LuxR based
system.
 Cytoplasmic receptor.
Brameyer, Sophie, Helge B. Bode, and Ralf Heermann. "Languages and dialects: bacterial communication beyond homoserine lactones."
Trends in microbiology 23.9 (2015)
9

Quorum sensing
circuits in
Pseudomonas
aeruginosa
 4 Autoinducer Synthases:
LasI, RhlI, PqsABCDH and
AmbBCDE
 4 Autoinducers:
C12-HSL,C4-HSL, PQS and IQS
 4 Receptors:
LasR, hlR, PqsR and ?
Papenfort, Kai, and Bonnie L. Bassler. "Quorum sensing signal-response systems in Gram-negative bacteria." Nature Reviews Microbiology
14.9 (2016)
10

Quorum sensing
inGram positive
bacteria
 Intraspecies communication.
 Peptide-mediated QS.
 Oligopeptide autoinducers.
 Receptors on cell membrane.
Camilli, Andrew, and Bonnie L. Bassler. "Bacterial small-molecule signaling pathways." Science 311.5764 (2006)
11

Staphylococcus
aureus quorum
sensing system
 ArgD:
pre-autoinducing peptide(AIP).
 ArgB:
trims ArgD into the functional.
AIP.
 ArgC:
receptor kinase.
 ArgA:
transcriptional activator.
. Madigan, Michael T., John M. Martinko, and Jack Parker. Brock biology of microorganisms.14th edition. 2014. 12

AI-2 system
 Gram negative and Gram
positive bacteria.
 Interspecies communication.
 (DPD) is synthesized by all
LuxS enzymes.
 AI-2, often in conjuncate with
an AHL or oligopeptide
autoinducer.
 Furanosyl / borate diester.
 Induces expression of
stationary phase genes.
Bacterial Esperanto !
Gobbetti, M., et al. "Cell–cell communication in food related bacteria." International journal of food microbiology 120.1 (2007) 13

14
History of quorum
sensing
Quorum sensing
in Prokaryotes
Quorum sensing
in Eukaryotes
Applications of
QS in medical
biotechnology
Applications of
QS in industrial
biotechnology
Applications of
QS in agricultural
biotechnology

Quorum sensing
in unicellular and
dimorphic fungi
 Farnesol
 Tryptophol
 Tyrosol
 Phenylethanol
Nickerson, Kenneth W., Audrey L. Atkin, and Jacob M. Hornby. "Quorum sensing in dimorphic fungi: farnesol and beyond." Applied and
environmental microbiology 72.6 (2006)
15

Quorum sensing
in filamentous
fungi
 Oxylipins
 Butyrolactones
Tsitsigiannis, Dimitrios I., and Nancy P. Keller. "Oxylipins as developmental and host–fungal communication signals."Trends in
microbiology 15.3 (2007) 16

QS applications
in biotechnology
Human
health
Industrial
products
Agriculture
17

18
History of quorum
sensing
Quorum sensing
in Prokaryotes
Quorum sensing
in Eukaryotes
Applications of
QS in medical
biotechnology
Applications of
QS in industrial
biotechnology
Applications of
QS in agricultural
biotechnology

Quorum quenching :
Silencing the majority !
Quorum quenching (QQ) is an alternative
antimicrobial therapeutics for human health.
QQ strategies :
1. QQ by small quorum-sensing inhibitors (QSI).
2. QQ by Enzymes.
Rasmussen, Thomas B., and Michael Givskov. "Quorum-sensing inhibitors as anti-pathogenic drugs." International Journal of Medical
Microbiology 296.2 (2006)
19

QS inhibitors
1) Antagonizing signal
binding to LuxR family
receptor.e.g,
Halogenatedacyl-
furanones.
2) Inhibition of signal
production.e.g,C8-HSL
analog.
3) Trapping signals.e.g,
Cyclodextrins.
4) Suppression of synthase
and receptor activities,
stabilities or productions.
e.g., QteE & cis-RNA.
Hirakawa, Hidetada, and HaruyoshiTomita. "Interference of bacterial cell-to-cell communication: a new concept of
antimicrobial chemotherapy breaks ant ibiotic resistance." Frontiers in microbiology 4 (2013).
20

QQ Enzymes
A. AHL-lactonase.e.g.,AiiA
from Bacillus spp.
B. AHL-acylase.e.g.,AiiC
from Anabaena spp.
C. AHL-oxidoreductases.eg.,
CYP102A1 from Bacillus
megaterium.
D. LsrK kinase.e.g., kinase
LsrK from E. coli.
Fetzner, Susanne. "Quorum quenching enzymes." Journal of biotechnology 201 (2015)
21

22
History of quorum
sensing
Quorum sensing
in Prokaryotes
Quorum sensing
in Eukaryotes
Applications of
QS in medical
biotechnology
Applications of
QS in industrial
biotechnology
Applications of
QS in agricultural
biotechnology

QS a novel strategy for
overproduction of
microbial products
Fungal secondary metabolism can improve with QS
molecules.
A European initiative worked to identify molecules involved
in this process in order to improve the productivity of
industrially important fungal bioproducts.
. http://cordis.europa.eu/result/rcn/88437_en.html
23

Sclerotiorin
overproduction
 overproduction of the
fungal antibiotic
sclerotiorin in cultures of
Penicillium sclerotiorum
(strain S) through the
addition of spent medium
from a culture of
Penicillium sclerotiorum
(strain M).
Raina, Sheetal, Mark Odell, andTajalli Keshavarz. "Quorum sensing as a method for improving sclerotiorin production in Penicillium
sclerotiorum." Journal of biotechnology 148.2 (2010)
24

increase in lovastatin
production and the
changes
in the transcription
levels in A.terreus
 lovastatin production of increase
2.5-fold increase in lovastatin
yield.
 addition of butyrolactone I at 96 h
caused an enhancement in the
levels of gene expression for both
lovB and lovF. increase 1.2-fold
lovB and 1.3-fold lovF amplicons
after 120 h of fermentation.
Raina, Sheetal, et al. "Is quorum sensing involved in lovastatin production in the filamentous fungus Aspergillus terreus?."
Process Biochemistry 47.5 (2012)
25

A.terreusQS
signalling
mechanism
1. 9-HpODE activate the
signalling mechanism
by binding to GPCRs.
2. Exchange of GDP to
GTP of the Gα subunit.
3. cAMP/ PKA signalling
pathways generation.
4. Expression of target
genes.
Amache, Rana (2014) Quorum sensing for improved production of industrially useful products from filamentous fungi. Doctoral thesis,
University of Westminster.
26

27
History of quorum
sensing
Quorum sensing
in Prokaryotes
Quorum sensing
in Eukaryotes
Applications of
QS in medical
biotechnology
Applications of
QS in industrial
biotechnology
Applications of
QS in agricultural
biotechnology

Quorum sensing
and agriculture
Positive
effects
Nitrogen-fixing
bacteria
Negative
effects
Plant
pathogens
28

QS effects on
Bacterium-
Plant
Symbiotic
Interactions
 The symbiotic relationships
formed between the nitrogen
fixing rhizobia and their
legume hosts are the result of
signaling networks between
the host and symbiont.
 Quorum sensing has been
implicated as a key player in
the symbiotic process.
quorum
sensing
nodulation
efficiency
exopolysa
ccharide
productio
n
nitrogen
fixation,
symbioso
me
developm
ent
González, Juan E., and Melanie M. Marketon. "Quorum sensing in nitrogen-fixing rhizobia." Microbiology and Molecular Biology
Reviews 67.4 (2003)
29

Phenomenon
regulated by
quorum sensing
System
responsible
for AHL
production
AHL(s) presentOrganism
Plasmid transfertraR/traI3-Oxo-C8-HSLA. tumefaciens
Nodulation
efficiency
rhiR/rhiI3-OH-C14:1-HSLR. leguminosarum
bv. viciae
Plasmid transfer,traR/traI3-Oxo-C8-HSLRhizobium sp. strain
NGR234
Exopolysaccharide
EPS II production
sinR/sinI3-Oxo-C14-HSLS. meliloti Rm41
Activation of nod
genes
UnknownBradyoxetinBradyrhizobium sp.
González, Juan E., and Melanie M. Marketon. "Quorum sensing in nitrogen-fixing rhizobia." Microbiology and Molecular
Biology Reviews 67.4 (2003)
30

QS as a
Biocontrol way
for plant heath
 Biocontrol strategies against
soft-rot bacteria.
 RhodococcalQuorum
Quenching vs. Pectobacterial
Quorum Sensing.
Latour, Xavier, et al. "Rhodococcus erythropolis and its γ-lactone catabolic pathway: an unusual biocontrol system
that disrupts pathogen quorum sensing communication." Agronomy 3.4 (2013)
31

Quorum
sensing in
agricultural
biotechnology
What can
biotechnologists
do?
Cirou, Amélie, et al. "Quorum sensing as a target for novel biocontrol strategies directed at Pectobacterium." Recent Developments in
Management of Plant Diseases. Springer Netherlands, 2010.
32

Our understanding of QS mechanisms is
still limited.
QS enzymes susceptibility to denaturation
and proteolytic degradation.
Most approaches are still in the research
phase.
33

Real-life applications.
Theoretical and practical aspects.
Chemical tools like biosensors.
Attractive application in food industry and human health.
Sustainable agriculture.
Test various edible mushroom extracts for fungal QSIs
Explore more QS activators for industrial biotechnology.
Quorum signaling impact on heterologous protein expression.
Improvement of the human intestinal flora. 34

35
Selected References:
1. Lee, Anna Jisu, and LingchongYou. "Cells Listen toTheir InnerVoice." Science 343.6171 (2014)
2. Brameyer, Sophie, Helge B. Bode, and Ralf Heermann. "Languages and dialects: bacterial
communication beyond homoserine lactones."Trends in microbiology 23.9 (2015)
3. Papenfort, Kai, and Bonnie L. Bassler. "Quorum sensing signal-response systems in Gram-negative
bacteria." Nature Reviews Microbiology 14.9 (2016)
4. Hirakawa, Hidetada, and HaruyoshiTomita. "Interference of bacterial cell-to-cell communication: a
new concept of antimicrobial chemotherapy breaks ant ibiotic resistance." Frontiers in microbiology
4 (2013)
5. Fetzner, Susanne. "Quorum quenching enzymes." Journal of biotechnology 201 (2015)
6. Raina, Sheetal, et al. "Is quorum sensing involved in lovastatin production in the filamentous fungus
Aspergillus terreus?." Process Biochemistry 47.5 (2012)
7. Latour, Xavier, et al. "Rhodococcus erythropolis and its γ-lactone catabolic pathway: an unusual
biocontrol system that disrupts pathogen quorum sensing communication." Agronomy 3.4 (2013)
8. Cirou, A., Uroz, S., et al. (2009) ‘Quorum Sensing as aTarget for Novel Biocontrol Strategies
Directed at Pectobacterium’, in RECENT DEVELOPMENTS IN MANAGEMENTOF PLANT
DISEASES. Editors: Gisi, Ulrich, Chet, I., Gullino, Maria Lodovica (Eds.) Dordrecht: Springer
Netherlands, pp. 121–131

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