Documente Academic
Documente Profesional
Documente Cultură
Year: 2009/2010
Biofouling refers to a process whereby unprotected artificial and natural substrata are
quickly colonised by biota in an aquatic environment (Railkin., 2004). Biofilm
formation, which can be mediated by QS, is the initial stage of biofouling (Dobretsov
et al., 2009) and it is therefore common to find compounds produced by marine
organisms, such as algae and sponges, which inhibits QS as a means of defence
against colonisation (Sauer et al., 2002). In addition, it is believed that sponges form a
symbiosis with microbes, such as bacteria and fungi (Kubanek et al., 2003) which
results in the production of many bioactive natural compounds, some of which were
shown to be potent QS inhibitors. Increasing research is confirming that new marine
microbes discovered among microbial communities in sponges are producers of
natural bioactive compounds (Wang., 2006). Therefore, screening for potential QS
inhibitors among the microbiota of marine sponges is a good strategy to identify QS
inhibitors which may be of potential therapeutic use in the future.
Figure 1. AHLs are synthesised by homologues of the AHL synthase LuxI from S-
adenosyl methionine and an intracellular pool of carrier proteins, with each AHL
distinguished by the length, degree of saturation, and substitution of the acyl side
chains (Parsek et al., 1999; Dobrestor et al., 2009). (Figure modified from Waters &
Bassler., 2005).
Figure 2. The red triangles indicate the autoinducer that is produced by LuxI. LuxI
and LuxR control the expression of a specific operon. LuxI functions as the
auotoinducer synthase which synthesises an AHL and the LuxR protein is an AHL-
responsive DNA binding transcriptional activator. After synthesis the AHL freely
diffuses in and out of the cell and the concentration increases as the cell density
increases. Upon reaching the threshold, AHL is bound by LuxR which initiates
transcription of the operon. A positive feedback loop is created, because the LuxR-
AHL complex also induces the expression of LuxI as it is encoded on the operon. This
floods the environment with the AHL signal causing the entire population to go into
―quorum sensing mode‖ (Kaplan & Greenberg., 1985; Stevens et al., 1994; Waters &
Bassler., 2005) (Figure from Waters & Bassler., 2005).
Screening strategies
A number of screening methods have been developed to screen for Quorum sensing
inhibitors (QSIs). A commonly used screen uses a QS-controlled promoter which is
fused to a reporter gene or operon, for example the violacin operon from
Chromobacterium violaceum. The reporter is activated when exogenous AHLs are
present and their corresponding reporters are activated. Exogenous QSI activity
reduces or fully inhibits expression of the reporters. A reduced signal, or no signal is a
positive result (Rasmussen et al., 2006). C. violaceum regulates pigment production
by C6-HSL which is inhibited by AHL analogues and other antagonists. A positive
result is indicated by the lack of pigmentation surrounding the QSI (McClean et al.,
2004).
An alternative screening method developed by Rasmussen and colleagues uses
genetically modified bacteria which are termed QSI selectors (QSIS) which are killed
if AHLs are present but survive if AHLs are present with a potential QSI (Rasmussen
et al., 2005a).
Conclusion
Quorum sensing and strategies to inhibit it have garnered increased attention in recent
years, especially as resistance to traditional antibiotics is at an all time high. The
attraction in using QS inhibitors against pathogens such as P. aeruginosa is that there
is less selective pressure for bacteria to mutate and develop resistance as the effect of
QS is not bactericidal, with the added benefit that there is no ―collateral damage‖ to
the normal microflora of the patient. Despite advances in the study of QS inhibitors,
there remains significant hurdles to be overcome before a QS inhibiting drug is
approved for therapeutic use. For example, it is unproven that QS inhibitors would
function in humans. In addition, some QS inhibitors, such as furanones and SAM
analogues have adverse side – effects (Wilcox et al., 2008). More mechanistic studies
of QS pathways is needed as their exact functional role is not clear (Ni et al., 2008). In
the search for QS inhibiting compounds, screening marine sponges is a good choice
due to the vast amount of bioactive compounds produced by the sponge microbiota.
Recent research in this area have recovered a large number of bacterial strains in
sponges which have antimicrobial effects, most by inhibiting QS. Because bacteria
rapidly produce large quantities of biomass, if potent QS inhibiting bacteria are
recovered, they can be grown on a biotechnological scale, and their bioactive
compounds harvested for use as antimicrobials (Marinho et al., 2009) once sufficient
optimisation/modification is achieved for tests on humans and subsequently approval.
To conclude, it is imperative for humans to find new antimicrobial agents as we are
currently losing the ―battle‖ against re - emerging infections due to antibiotic
resistance. Using QS inhibiting bioactive compounds produced by bacteria is
therefore an important strategy in developing alternative treatments for infections
caused by multidrug – resistant bacteria.
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