Three CoA Transferases Involved in the Production of Short Chain Fatty Acids in Porphyromonas gingivalis

Abstract

Butyryl-CoA:acetate CoA transferase, which produces butyrate and acetyl-CoA from butyryl-CoA and acetate, is responsible for the final step of butyrate production in bacteria. This study demonstrates that in the periodontopathogenic bacterium Porphyromonas gingivalis this reaction is not catalyzed by PGN_1171, previously annotated as butyryl-CoA:acetate CoA transferase, but by three distinct CoA transferases, PGN_0725, PGN_1341, and PGN_1888. Gas chromatography/mass spectrometry (GC-MS) and spectrophotometric analyses were performed using crude enzyme extracts from deletion mutant strains and purified recombinant proteins. The experiments revealed that, in the presence of acetate, PGN_0725 preferentially utilized butyryl-CoA rather than propionyl-CoA. By contrast, this preference was reversed in PGN_1888. The only butyryl-CoA:acetate CoA transferase activity was observed in PGN_1341. Double reciprocal plots revealed that all the reactions catalyzed by these enzymes follow a ternary-complex mechanism, in contrast to previously characterized CoA transferases. GC-MS analysis to determine the concentrations of short chain fatty acids (SCFAs) in culture supernatants of P. gingivalis wild type and mutant strains revealed that PGN_0725 and PGN_1888 play a major role in the production of butyrate and propionate, respectively. Interestingly, a triple deletion mutant lacking PGN_0725, PGN_1341, and PGN_1888 produced low levels of SCFAs, suggesting that the microorganism contains CoA transferase(s) in addition to these three enzymes. Growth rates of the mutant strains were mostly slower than that of the wild type, indicating that many carbon compounds produced in the SCFA synthesis appear to be important for the biological activity of this microorganism.

Global analysis of lysine succinylome in the periodontal pathogen Porphyromonas gingivalis

“…Butyrate and propionate are known to easily penetrate the periodontal tissue and subsequently disturb host defense systems. It has been reported that concentrations of these molecules in the periodontal pockets are significantly correlated with disease severity and inflammation . It is worthwhile to note that, as shown in Figure , these vital catabolic pathways show 15 succinylated enzymes.…”

“…Proposed metabolic map of energy and cytotoxin production in P gingivalis . This scheme was constructed based on previous reports . Catalytic enzymes are represented by their P gingivalis PGN designation.…”

The concordance between upper and lower respiratory microbiota in children with Mycoplasma pneumoniae pneumonia

“…In contrast to the two NP groups identified, the OP microbiota in healthy children showed no significant difference. The respiratory commensals decreased significantly in MPP, including a reduced amount of the lactic acid-producing bacterium Lactobacillus and the short-chain fatty acid producer Porphyromonas 19 , 20 . Prevotella , which antagonizes lipid polysaccharides and inhibits mucosal inflammation, was also decreased 21 .…”

Sinus Microanatomy and Microbiota in a Rabbit Model of Rhinosinusitis

“…In the gastrointestinal tract, carbon-source nutrients (e.g., butyrate) produced by bacteria are thought to play an important and beneficial role (Consolandi et al, 2015 ). However, carbon-source nutrients that induce apoptosis in fibroblasts and lymphocytes (T- and B-cells) can be the toxic metabolic end product in the airway (Chang et al, 2013 ; Flynn et al, 2016 ; Sato et al, 2016 ). Despite the high density of bacteria that colonize the airway, nutrient sources that sustain bacterial growth in vivo , and how those nutrients are derived, are not well-characterized and the current rabbit model is well-suited to further elucidate such mechanism responsible for the generation of CRS secondary to pathogenic bacteria.…”