Dataset Information

Proteome response of biofilm forming Pseudomonas fluorescens ITEM 17298 to pepsin-digested bovine lactoferrin under two incubation temperatures

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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Publications

Proteomic analysis of the food spoiler Pseudomonas fluorescens ITEM 17298 reveals the antibiofilm activity of the pepsin-digested bovine lactoferrin.

Quintieri Laura L Zühlke Daniela D Fanelli Francesca F Caputo Leonardo L Liuzzi Vania Cosma VC Logrieco Antonio Francesco AF Hirschfeld Claudia C Becher Dörte D Riedel Katharina K

Food microbiology 20190208

Pseudomonas fluorescens is implicated in food spoilage especially under cold storage. Due to its ability to form biofilm P. fluorescens resists to common disinfection strategies increasing its persistance especially across fresh food chain. Biofilm formation is promoted by several environmental stimuli, but gene expression and protein changes involved in this lifestyle are poorly investigated in this species. In this work a comparative proteomic analysis was performed to investigate metabolic pa ...[more]

PMID: 31027772

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Project description:Biofilm formation on medically implanted devices by Candida albicans poses a significant clinical challenge. Here we compared biofilm-associated gene expression in two clinical C. albicans isolates, SC5314 and WO-1, to identify shared gene regulatory responses that may be functionally relevant. Among the 50 genes most highly expressed in biofilms relative to planktonic (suspension-grown) cells, we were able to recover insertion mutations in 25 genes. We observed that 20 of the 25 mutants have altered biofilm-related properties, including cell-substrate adherence, cell-cell signaling, and azole susceptibility. We focused on the most highly up-regulated gene in biofilms, RHR2, which specifies the glycerol biosynthetic enzyme glycerol-3-phosphate phosphatase. Glycerol is 5-fold more abundant in biofilm cells than planktonic cells, and an rhr2D/D strain accumulates 2-fold less biofilm glycerol than the wild type. Under in vitro growth conditions, the rhr2D/D mutant has reduced biofilm biomass and reduced adherence to silicone. The rhr2D/D mutant is severely defective in biofilm formation in vivo, in a rat catheter infection model. Expression profiling of the rhr2D/D mutant indicates that it has reduced expression of cell surface adhesin genes ALS1, ALS3, and HWP1, as well as a large fraction of all other biofilm up-regulated genes. Reduced adhesin expression is the cause of the rhr2D/D mutant biofilm defect, because overexpression of ALS1, ALS3, or HWP1 restores biofilm formation ability to the mutant in vitro and in vivo. Our findings indicate that internal glycerol has a regulatory role in biofilm gene expression, and that adhesin genes are among the main functional Rhr2-regulated genes. Gene expression profiles, in duplicate; (1) for biofilm vs. planktonic growth conditions for the two wild-type clinical isolates of Candida albicans (SC5314 and WO1-white/WO1-opaque), and (2) for rhr2M-NM-^T/M-NM-^T mutant and complemented strain, via RNA-deep sequencing using Illumina GA2 and HiSeq2000 platforms, respectively

Dataset Information

Proteome response of biofilm forming Pseudomonas fluorescens ITEM 17298 to pepsin-digested bovine lactoferrin under two incubation temperatures

Publications

Proteomic analysis of the food spoiler Pseudomonas fluorescens ITEM 17298 reveals the antibiofilm activity of the pepsin-digested bovine lactoferrin.

Similar Datasets

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