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ROLES OF RESPIRATORY CHAIN ENZYMES IN BIOFILM FORMATION OF PSEUDOMONAS AERUGINOSA
Title
ROLES OF RESPIRATORY CHAIN ENZYMES IN BIOFILM FORMATION OF PSEUDOMONAS AERUGINOSA
Creator
DING, JIE
Date
2019
Description
Pseudomonas aeruginosa is a Gram-negative, rod-shaped bacterium, resistant to many antibiotics. It can cause several chronic infections such...
Show morePseudomonas aeruginosa is a Gram-negative, rod-shaped bacterium, resistant to many antibiotics. It can cause several chronic infections such as lung, bloodstream, urinary tract, and surgical wound infections. This bacterium produces biofilms which confer resistance to hazardous environments. P. aeruginosa contains five stages of colony development, which are planktonic attachment, cell to cell adhesion, proliferation, maturation, and dispersion. After five stages, biofilms of P. aeruginosa are matured. The biofilm structure produced by P. aeruginosa is important for cell survival, providing protection and resistance to harsh environment and antibiotics. In this research, the biofilms formed by wildtype strain PA01 and mutated strains of PA14, including NDH-2, NQR F, and NUO I, were developed in LB medium and Artificial Urine Medium separately for 96 hours. After washing, collecting, and staining the biofilms, the analyses of measurement of OD562 showed that in LB medium, PA01 formed more biofilms than mutants while NUO I and NDH2 had less biofilms, although not significantly. In AUM the situation was different. PA01 formed least biofilms while NQR F formed largest biofilms than any other strains. Also, the NDH-2 formed more biofilms than NUO I in AUM. The deficiencies of enzymes loss in those strains result in growing biofilm concentrations. Because the difference was not significant, we can only say that the NQR and NADH dehydrogenases have important roles in biofilm formation.
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DEVELOPING FUSION BACTERIOCINS FOR ERADICATING PSEUDOMONAS AERUGINOSA BIOFILMS
Title
DEVELOPING FUSION BACTERIOCINS FOR ERADICATING PSEUDOMONAS AERUGINOSA BIOFILMS
Creator
An, Sungjun
Date
2022
Description
The opportunistic pathogen Pseudomonas aeruginosa is a leading cause of morbidity and mortality in cystic fibrosis patients and...
Show moreThe opportunistic pathogen Pseudomonas aeruginosa is a leading cause of morbidity and mortality in cystic fibrosis patients and immunocompromised individuals. Due to its remarkable ability to resist antibiotics, eradicating P. aeruginosa has become increasingly difficult. As previously reported, we have successfully engineered a colicin-secretion system that kills target biofilm cells rapidly and selectively in multispecies biofilms as well as demonstrated the potential of using live microorganisms engineered to produce antimicrobial colicin protein to treat biofilm-associated infections. In this study,we constructed a fusion colicin-pyocin that could target P. aeruginosa by DNase activity of colicin E2. The newly engineered bacteriocin-secretion system upon the shift in target, maintained biofilm inhibition capacity. Both during biofilm formation and after its development, the system was able to suppress the P. aeruginosa biofilm. This result opened up the possibility that it could be used for novel live biotherapeutics. A further study was conducted to overcome the challenge of requiring an exogenous inducer. We applied the concept of Quorum-Sensing signal that recognize autoinducer as a trigger of fusion colicin-pyocin producing genetic circuit so that it automates the production and secretion of fusion colicin-pyocin as soon as the genetic circuit senses the target population growing. This study demonstrated that combining the domains of colicin and pyocin could broaden the genetic circuit target range, maintaining strain specificity, while employing the QS system could remove the fundamental problem of diffusion or degradation of extra compounds as they approach engineered cells.
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