Proteome response of biofilm forming Pseudomonas fluorescens ITEM 17298 to pepsin-digested bovine lactoferrin under two incubation temperatures
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ABSTRACT: Pseudomonas fluorescens is implicated in food spoilage especially under cold storage, causing the increase of food waste and the reduction of environmental (and nutritional) sustainability. In food industry, Pseudomonas biofilm on pipe surfaces and facilities creates a persistent source of contamination and increases resistance to common disinfection strategies. Even though gene expression and protein pattern changes under environmental stimuli have been suggested, cellular mechanisms involved in biofilm production are poorly investigated for this species. In the present work, the cheese blue pigmenting P. fluorescens ITEM 17298 produced increased biofilm biomass and motility at 15 °C in comparison to 30 °C. Proteomic analysis revealed major differences of proteins involved in biofilm formation. In particular, at 15 °C we found increased amount of the c-di-GMP regulator PleD involved in motility as well as AlgB and the carbon storage regulator directly involved biofilm formation. Carbon metabolism, protein and fatty acid synthesis were also stimulated under cold incubation. Moreover, for the first time, enzymes correlated to indigoidine synthesis (blue pigment) were identified in the same condition. The differences in expression of regulatory proteins could therefore be the major determinant of the biofilm formation ability of P. fluorescens under refrigerated condition. Genomic and proteomic results also revealed virulence factors mostly detected at 30 °C. In order to counteract P. fluorescens persistence, this study also proposed a control strategy for biofilm development. Thus, a sub-lethal concentration of bovine lactoferrin hydrolysate (HLF) was assayed at both temperatures. Treatment caused a significant reduction in biofilm biomass, also confirmed by changes in proteome pattern. At each condition proteins involved regulatory functions and transcription factors were downregulated after HLF-treatment. Among these, at 15 °C we found that PleD was absent in treated samples, whilst negative regulators of alginate biosynthesis were detected; likewise, indigoidine related proteins were repressed. The cyclic-di-GMP-binding biofilm dispersal mediator was instead detected in all treated samples.
INSTRUMENT(S): LTQ Orbitrap Velos
ORGANISM(S): Pseudomonas Fluorescens Group
SUBMITTER: Daniela Zuehlke
LAB HEAD: Kathrin Riedel
PROVIDER: PXD010477 | Pride | 2019-05-28
REPOSITORIES: Pride
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