1887

Abstract

secretes a variety of hydrolases, many of which contribute to virulence or are thought to play a role in the nutrition of the bacterium. As most studies concerning extracellular enzymes have been performed on planktonic cultures of non-mucoid strains, knowledge of the potential role of these enzymes in biofilm formation in mucoid (alginate-producing) remains limited. Here we show that mucoid produces extracellular hydrolases during biofilm growth. Overexpression of the extracellular lipases LipA and LipC, the esterase EstA and the proteolytic elastase LasB from plasmids revealed that some of these hydrolases affected the composition and physicochemical properties of the extracellular polymeric substances (EPS). While no influence of LipA was observed, the overexpression of and led to increased concentrations of extracellular rhamnolipids with enhanced levels of mono-rhamnolipids, elevated amounts of total carbohydrates and decreased alginate concentrations, resulting in increased EPS hydrophobicity and viscosity. Moreover, we observed an influence of the enzymes on cellular motility. Overexpression of resulted in a loss of twitching motility, although it enhanced the ability to swim and swarm. The -overexpression strain showed an overall enhanced motility compared with the parent strain. Moreover, the EstA- and LasB-overproduction strains completely lost the ability to form 3D biofilms, whereas the overproduction of LipC increased cell aggregation and the heterogeneity of the biofilms formed. Overall, these findings indicate that directly or indirectly, the secreted enzymes EstA, LasB and LipC can influence the formation and architecture of mucoid biofilms as a result of changes in EPS composition and properties, as well as the motility of the cells.

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2019-10-14
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Enzyme activities in EPS-containing cell-free supernatants from LB cultures of the mucoid strain SG81 and its derivative strains [PDF](13 KB)

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Gel electrophoretic analysis of PCR fragments amplified from genomic DNA from SG81 in 0.8% (w/v) agarose gels [PDF](389 KB)

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