The pyruvate oxidase (SpxB)–dependent production of H2O2 is widely distributed among oral commensal streptococci. Several studies confirmed the ability of H2O2 to antagonize susceptible oral bacterial species, including caries-associated Streptococcus mutans as well as several periodontal pathobionts. Here we report a potential mechanism to bolster oral commensal streptococcal H2O2 production by magnesium (Mg2+) supplementation. Magnesium is a cofactor for SpxB catalytic activity, and supplementation increases the production of H2O2 in vitro. We demonstrate that Mg2+ affects spxB transcription and SpxB abundance in Streptococcus sanguinis and Streptococcus gordonii. The competitiveness of low-passage commensal streptococcal clinical isolates is positively influenced in antagonism assays against S. mutans. In growth conditions normally selective for S. mutans, Mg2+ supplementation is able to increase the abundance of S. sanguinis in dual-species biofilms. Using an in vivo biophotonic imaging platform, we further demonstrate that dietary Mg2+ supplementation significantly improves S. gordonii oral colonization in mice. In summary, our results support a role for Mg2+ supplementation as a potential prebiotic to promote establishment of oral health–associated commensal streptococci.
BarnardJPStinsonMW. 1999. Influence of environmental conditions on hydrogen peroxide formation by Streptococcus gordonii. Infect Immun. 67(12):6558–6564.
2.
BowenWHBurneRAWuHKooH. 2018. Oral biofilms: pathogens, matrix, and polymicrobial interactions in microenvironments. Trends Microbiol. 26(3):229–242.
3.
ConstantopoulosGBarrangerJA. 1984. Nonenzymatic decarboxylation of pyruvate. Anal Biochem. 139(2):353–358.
4.
DamoSMKehl-FieTESugitaniNHoltMERathiSMurphyWJZhangYBetzCHenchLFritzG, et al. 2013. Molecular basis for manganese sequestration by calprotectin and roles in the innate immune response to invading bacterial pathogens. Proc Natl Acad Sci USA. 110(10):3841–3846.
5.
GroismanEAHollandsKKrinerMALeeEJParkSYPontesMH. 2013. Bacterial Mg2+ homeostasis, transport, and virulence. Annu Rev Genet. 47:625–646.
6.
HaleJDTingYTJackRWTaggJRHengNC. 2005. Bacteriocin (mutacin) production by Streptococcus mutans genome sequence reference strain UA159: elucidation of the antimicrobial repertoire by genetic dissection. Appl Environ Microbiol. 71(11):7613–7617.
7.
HeJHwangGLiuYGaoLKilpatrick-LivermanLSantarpiaPZhouXKooH. 2016. L-arginine modifies the exopolysaccharide matrix and thwarts Streptococcus mutans outgrowth within mixed-species oral biofilms. J Bacteriol. 198(19):2651–2661.
8.
HerreroERSlomkaVBernaertsKBoonNHernandez-SanabriaEPassoniBBQuirynenMTeughelsW. 2016. Antimicrobial effects of commensal oral species are regulated by environmental factors. J Dent. 47:23–33.
9.
HossainMSBiswasI. 2011. Mutacins from Streptococcus mutans UA159 are active against multiple streptococcal species. Appl Environ Microbiol. 77(7):2428–2434.
10.
HuangXBrowngardtCMJiangMAhnSJBurneRANascimentoMM. 2018. Diversity in antagonistic interactions between commensal oral streptococci and Streptococcus mutans. Caries Res. 52(1–2):88–101.
11.
ItzekAZhengLChenZMerrittJKrethJ. 2011. Hydrogen peroxide-dependent DNA release and transfer of antibiotic resistance genes in Streptococcus gordonii. J Bacteriol. 193(24):6912–6922.
12.
JakubovicsNSGillSRVickermanMMKolenbranderPE. 2008. Role of hydrogen peroxide in competition and cooperation between Streptococcus gordonii and Actinomyces naeslundii. FEMS Microbiol Ecol. 66(3):637–644.
13.
JenCMShelefLA. 1986. Factors affecting sensitivity of Staphylococcus aureus 196E to polyphosphates. Appl Environ Microbiol. 52(4):842–846.
14.
KrethJGiacamanRARaghavanRMerrittJ. 2017. The road less traveled—defining molecular commensalism with Streptococcus sanguinis. Mol Oral Microbiol. 32(3):181–196.
15.
KrethJVuHZhangYHerzbergMC. 2009. Characterization of hydrogen peroxide-induced DNA release by Streptococcus sanguinis and Streptococcus gordonii. J Bacteriol. 191(20):6281–6291.
16.
KrethJZhangYHerzbergMC. 2008. Streptococcal antagonism in oral biofilms: Streptococcus sanguinis and Streptococcus gordonii interference with Streptococcus mutans. J Bacteriol. 190(13):4632–4640.
17.
LiuYBeyerAAebersoldR. 2016. On the dependency of cellular protein levels on mRNA abundance. Cell. 165(3):535–550.
18.
MarshPDHeadDADevineDA. 2015. Ecological approaches to oral biofilms: control without killing. Caries Res. 49(Suppl 1):46–54.
19.
MarshPDZauraE. 2017. Dental biofilm: ecological interactions in health and disease. J Clin Periodontol. 44(Suppl 18):S12–S22.
20.
MerrittJSenpukuHKrethJ. 2016. Let there be bioluminescence: development of a biophotonic imaging platform for in situ analyses of oral biofilms in animal models. Environ Microbiol. 18(1):174–190.
21.
MiraASimon-SoroACurtisMA. 2017. Role of microbial communities in the pathogenesis of periodontal diseases and caries. J Clin Periodontol. 44(Suppl 18):S23–S38.
22.
MonaciFBargagliEBraviFRottoliP. 2002. Concentrations of major elements and mercury in unstimulated human saliva. Biol Trace Elem Res. 89(3):193–203.
23.
NascimentoMMBrowngardtCXiaohuiXKlepac-CerajVPasterBJBurneRA. 2014. The effect of arginine on oral biofilm communities. Mol Oral Microbiol. 29(1):45–54.
24.
NudlerE. 2009. RNA polymerase active center: the molecular engine of transcription. Annu Rev Biochem. 78:335–361.
25.
QiFChenPCaufieldPW. 2001. The group I strain of Streptococcus mutans, UA140, produces both the lantibiotic mutacin I and a nonlantibiotic bacteriocin, mutacin IV. Appl Environ Microbiol. 67(1):15–21.
RedanzSChengXGiacamanRAPfeiferCSMerrittJKrethJ. 2018. Live and let die: hydrogen peroxide production by the commensal flora and its role in maintaining a symbiotic microbiome. Mol Oral Microbiol. 33(5):337–352.
28.
RedanzSMasilamaniRCullinNGiacamanRAMerrittJKrethJ. 2018. Distinct regulatory role of carbon catabolite protein a (CcpA) in oral streptococcal spxB expression. J Bacteriol. 200(8). pii: e00619-17.
29.
SaitoMSekiMIidaKNakayamaHYoshidaS. 2007. A novel agar medium to detect hydrogen peroxide-producing bacteria based on the Prussian blue-forming reaction. Microbiol Immunol. 51(9):889–892.
30.
SchlaferSMeyerRLDigeIReginaVR. 2017. Extracellular DNA contributes to dental biofilm stability. Caries Res. 51(4):436–442.
SedewitzBSchleiferKHGotzF. 1984. Purification and biochemical characterization of pyruvate oxidase from Lactobacillus plantarum. J Bacteriol. 160(1):273–278.
33.
SkjorlandKGjermoPRollaG. 1978. Effect of some polyvalent cations on plaque formation in vivo. Scand J Dent Res. 86(2):103–107.
34.
StacyAEverettJJorthPTrivediURumbaughKPWhiteleyM. 2014. Bacterial fight-and-flight responses enhance virulence in a polymicrobial infection. Proc Natl Acad Sci USA. 111(21):7819–7824.
35.
TakahashiNNyvadB. 2011. The role of bacteria in the caries process: ecological perspectives. J Dent Res. 90(3):294–303.
36.
TittmannKWilleGGolbikRWeidnerAGhislaSHubnerG. 2005. Radical phosphate transfer mechanism for the thiamin diphosphate- and FAD-dependent pyruvate oxidase from Lactobacillus plantarum: kinetic coupling of intercofactor electron transfer with phosphate transfer to acetyl-thiamin diphosphate via a transient FAD semiquinone/hydroxyethyl-ThDP radical pair. Biochemistry. 44(40):13291–13303.
37.
VogelCMarcotteEM. 2012. Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet. 13(4):227–232.
38.
WhitchurchCBTolker-NielsenTRagasPCMattickJS. 2002. Extracellular DNA required for bacterial biofilm formation. Science. 295(5559):1487.
39.
ZengLMartinoNCBurneRA. 2012. Two gene clusters coordinate galactose and lactose metabolism in Streptococcus gordonii. Appl Environ Microbiol. 78(16):5597–5605.
40.
ZhengLYItzekAChenZYKrethJ. 2011. Oxygen dependent pyruvate oxidase expression and production in Streptococcus sanguinis. Int J Oral Sci. 3(2):82–89.
41.
ZhouPLiXHuangIHQiF. 2017. Veillonella catalase protects the growth of Fusobacterium nucleatum in microaerophilic and Streptococcus gordonii–resident environments. Appl Environ Microbiol. 83(19). pii: e01079-17.
42.
ZhouXXuXLiJHuDHuTYinWFanYZhangX. 2018. Oral health in China: from vision to action. Int J Oral Sci. 10(1):1.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
