Abstract
BackgroundHydrogen sulfide (H2S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease. We studied the H2S-producing protein expression of bacterial strains associated with periodontal disease. Further, we examined the effect of a cysteine-rich growth environment on the synthesis of intracellular enzymes in F. nucleatum polymorphum ATCC 10953. The proteins were subjected to one-dimensional (1DE) and two-dimensional (2DE) gel electrophoresis An in-gel activity assay was used to detect the H2S-producing enzymes; Sulfide from H2S, produced by the enzymes in the gel, reacted with bismuth forming bismuth sulfide, illustrated as brown bands (1D) or spots (2D) in the gel. The discovered proteins were identified with liquid chromatography – tandem mass spectrometry (LC-MS/MS).ResultsCysteine synthase and proteins involved in the production of the coenzyme pyridoxal 5′phosphate (that catalyzes the production of H2S) were frequently found among the discovered enzymes. Interestingly, a higher expression of H2S-producing enzymes was detected from bacteria incubated without cysteine prior to the experiment.ConclusionsNumerous enzymes, identified as cysteine synthase, were involved in the production of H2S from cysteine and the expression varied among Fusobacterium spp. and strains. No enzymes were detected with the in-gel activity assay among the other periodontitis-associated bacteria tested. The expression of the H2S-producing enzymes was dependent on environmental conditions such as cysteine concentration and pH but less dependent on the presence of serum and hemin.
Highlights
Hydrogen sulfide (H2S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease
H2S-producing enzymes among bacterial strains Cell extracts of 14 strains of bacteria associated with periodontitis (Table 1) were screened for H2S-producing enzymes with in-gel activity assay after renaturation
In this study we focused on bacterial production of H2S from cysteine
Summary
Hydrogen sulfide (H2S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease. The biofilms occupying the periodontal pocket, the area between the tooth and the surrounding connective tissue, are usually dominated by Gram-positive, facultative anaerobic bacteria but can undergo a compositional change towards Gram-negative, anaerobic and motile bacteria when oral hygiene is insufficient [1] The latter biofilms utilize the gingival crevicular fluid as a nutrient source and metabolize proteins, peptides and amino acids to various carboxylic acids and volatile sulfur compounds (VSC). Hydrogen sulfide (H2S) is the most common VSC formed by bacterial degradation of mainly the sulfurcontaining amino acid cysteine in the oral cavity It is a low-molecular weight and volatile gas compound detected in halitosis (bad breath) patients and in periodontal pockets in patients with periodontitis [2,3,4].
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