- Mutational analysis on stable expression and LasB inhibition of LasB propeptide in Pseudomonas aeruginosa
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Youngsun Shin , Xi-Hui Li , Cheol Seung Lee , Joon-Hee Lee
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J. Microbiol. 2022;60(7):727-734. Published online May 25, 2022
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DOI: https://doi.org/10.1007/s12275-022-1671-5
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 Abstract
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Three major proteases, elastase B (LasB), protease IV (PIV),
and elastase A (LasA) expressed in Pseudomonas aeruginosa
play important roles in infections and pathogeneses. These
are activated by a proteolytic cascade initiated by the activation
of LasB. In this study, we investigated whether LasB
could be inhibited using its propeptide (LasBpp). Although
LasA and PIV were inhibited by their propeptides, LasB was
not inhibited by purified LasBpp because LasB degraded LasBpp.
To address this problem, mutant LasBpp variants were constructed
to obtain a mutant LasBpp resistant to LasB degradation.
A C-terminal deletion series of LasBpp was tested in
vivo, and two positive candidates, T2 and T2-1, were selected.
However, both caused growth retardation and were unstably
expressed in vivo. Since deleting the C-terminal end of LasBpp
significantly affected its stable expression, substitution mutations
were introduced at the two amino acids near the
truncation site of T2-1. The resulting mutants, LasBppE172D,
LasBppG173A, and LasBppE172DG173A, significantly diminished LasB
activity when overexpressed in vivo and were stably expressed
in MW1, a quorum sensing mutant that does not produce
LasB. In vitro analysis showed that purified LasBppE172DG173A
inhibited LasB activity to a small extent. Summarizing, Cterminal
modification of LasBpp profoundly affected the stable
expression of LasBpp, and little enhanced the ability of
LasBpp to resist degradation by LasB.
- Antibiofilm effect of biofilm-dispersing agents on clinical isolates of Pseudomonas aeruginosa with various biofilm structures
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Soo-Kyoung Kim , Xi-Hui Li , Hyeon-Ji Hwang , Joon-Hee Lee
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J. Microbiol. 2018;56(12):902-909. Published online October 25, 2018
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DOI: https://doi.org/10.1007/s12275-018-8336-4
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75
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Abstract
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Pseudomonas aeruginosa, an opportunistic human pathogen,
causes many biofilm-mediated chronic infections. In this study,
biofilm structures of various clinical strains of P. aeruginosa
isolated from hospitalized patients were examined and their
influence on the biofilm-dispersing effects of chemicals was
investigated. The clinical isolates formed structurally distinct
biofilms that could be classified into three different groups:
1) mushroom-like, 2) thin flat, and 3) thick flat structures.
A dispersion of these differently structured biofilms was induced
using two biofilm-dispersing agents, anthranilate and
sodium nitroprusside (SNP). Although both SNP and anthranilate
could disperse all types of biofilms, the thick flat biofilms
were dispersed less efficiently than the biofilms of other
structures. This suggests that biofilm-dispersing agents have
higher potency on the biofilms of porous structures than on
densely packed biofilms.
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- [Minireview] Antibiofilm agents: A new perspective for antimicrobial strategy
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Xi-Hui Li , Joon-Hee Lee
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J. Microbiol. 2017;55(10):753-766. Published online September 28, 2017
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DOI: https://doi.org/10.1007/s12275-017-7274-x
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133
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127
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Abstract
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Biofilms are complex microbial architectures that attach to
surfaces and encase microorganisms in a matrix composed
of self-produced hydrated extracellular polymeric substances
(EPSs). In biofilms, microorganisms become much more
resistant to antimicrobial treatments, harsh environmental
conditions, and host immunity. Biofilm formation by microbial
pathogens greatly enhances survival in hosts and causes
chronic infections that result in persistent inflammation and
tissue damages. Currently, it is believed over 80% of chronic
infectious diseases are mediated by biofilms, and it is known
that conventional antibiotic medications are inadequate at
eradicating these biofilm-mediated infections. This situation
demands new strategies for biofilm-associated infections,
and currently, researchers focus on the development of antibiofilm
agents that are specific to biofilms, but are nontoxic,
because it is believed that this prevents the development of
drug resistance. Here, we review the most promising antibiofilm
agents undergoing intensive research and development.
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