Abstract
Bacterial biofilms are the bacterial aggregates that are embedded in the self-produced matrix of extracellular polymeric substances (EPS) that cause persistent bacterial infections posing significant medical challenges. They are recalcitrant to antibiotics and host defenses which make the treatments difficult and costly. Penicillium janthinellum mutant EU2D-21 was found to produce extracellular enzyme complex (amylase, cellulase, protease) under submerged fermentation. Maximum specific enzyme activities were found to be 3.04 IU/mg, 2.61 IU/mg and 3.39 IU/mg for alpha-amylase, cellulase and protease respectively, after 8 days of incubation at 30˚C. We evaluated the enzyme complex for its ability to target and degrade the biofilms of different bacteria. We found that it degraded biofilms of Escherichia coli (85.5%), Salmonella enterica (79.72%), Pseudomonas aeruginosa (88.76%) and Staphyloccus aureus (87.42%) within 1 h of incubation at 50˚C. The scanning electron microscopy (SEM), quantitation of biofilm removal assay and Crystal violet assay demonstrated that the enzyme complex detached the biofilm exo-polysaccharide matrix and bacteria from the cell surface. These results illustrate the feasibility and benefits of using this enzyme complex as anti-biofilm therapeutics to eradicate biofilms. This can also be used as a promising strategy to improve treatment of multidrug resistant bacterial infections.
Highlights
Our results suggest that the enzyme complex produced by P. janthinellum have the potential to effectively degrade bacterial biofilm and can act as an anti-biofilm therapeutics which can be used for treatment and eradication of chronic bacterial infections
Bacterial biofilms are single or multispecies microbial communities that are composed of cells embedded in self-produced extracellular polymeric substances (EPS)
These biofilms protect the embedded bacteria from sheer stress, disinfectants, host immune defenses and antibiotics which lead to perpetual bacterial infections
Summary
It is estimated that most of the human bacterial infections (60% - 80%) are due to biofilms [5] and there is an urgent need to develop novel and effective treatments that target and disrupt biofilms. Biofilms are resistant to antimicrobial agents due to the impaired penetration and it is necessary to develop prophylactic treatments that either inhibit biofilm formation [6] [7] or disrupt the biofilms [8]. Recent studies have shown great interest in the use of enzymes in targeting and disrupting the bacterial biofilms proving their potential in treatment and eradication of chronic bacterial diseases. The fungal strain, Aspergillus clavatus MTCC 1323 was found to produce enzyme complex (protease, amylase, pectinase) which degrades the biofilms of P. aeruginosa, B. subtilis and S. aureus [11]. Blanchette and Wenke [13] have very nicely highlighted the current and upcoming therapies for prevention and disruption of bacterial biofilms in their review article
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