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GNPS oral in vitro biofilms

ABSTRACT: These biofilms were established from saliva samples as described in Edlund et al. 2013 (Microbiome, doi:10.1186/2049-2618-1-25). After starving the biofilms in minimal medium anaerobiaclly glucose was added to the cultures. Samples for metatranscriptomics and metabolomics were collected in parallel at pH 7 (prior to glucose amendment), pH 4.2 (6 hours after glucose amendment) and pH 5.2 (9 hours after pH amendment). Secreted small molecules were extracted from biofilms (cells were not lysed) at the differnet pH stages.

INSTRUMENT(S): LTQ FT

ORGANISM(S): Human Oral Microbial Flora

SUBMITTER: Anna Edlund Jeffrey McLean

PROVIDER: MSV000078579 | GNPS | Thu Apr 10 15:18:00 BST 2014

REPOSITORIES: GNPS

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Project description:Candida albicans, a major opportunistic fungal pathogen is frequently found together with Streptococcus mutans in dental biofilms associated with severe childhood tooth-decay, a prevalent pediatric oral disease. Previous studies have demonstrated that S. mutans and C. albicans synergizes virulence of plaque-biofilms in vivo. However, the nature of this bacterial-fungal relationship in this cross-kingdom biofilm remains largely uncharacterized. Using iTRAQ based quantitative proteomics, we found that proteins associated with carbohydrate metabolism such as alpha-1,4 glucan phosphorylase, Hexokinase-2, Isocitrate lyase and malate synthase were significantly upregulated in C. albicans in the mixed-species biofilms (P<0.05). C. albicans proteins associated with growth/morphogenesis such as pH-responsive protein-2, Fma1p and Hsp21 were also induced. Conversely, S. mutans proteins in the tricarboxylic acid cycle such as citrate synthase and in the pentose phosphate pathway such as Ribose-5-phosphate isomerase A as well as proteins associated with sugar transport systems were upregulated indicating enhanced carbohydrate metabolism. Interestingly mixed-species biofilm microenvironment had a lower pH than S. mutans single-species biofilms. This observation was supported by proteomics, wherein proteins associated with lactate and formate assimilation such as Glyoxalase and putative NADPH-dependent methylglyoxal reductase proteins were significantly upregulated in the mixed-species biofilms (P<0.05). Furthermore, we unexpectedly found that S. mutans derived glucosyltransferase B (GtfB), responsible for co-adhesion via glucans, can also contribute to C. albicans growth and carbohydrate metabolism by providing glucose and fructose from sucrose breakdown. These findings demonstrate synergistic bacterial-fungal interactions within mixed-species biofilms and a novel GtfB cross-feeding role. Taken together, quantitative proteomics provides new insights into this virulent cross-kingdom oral biofilm.

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GNPS oral in vitro biofilms

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