Preliminary analysis in functions of clinical <i>Streptococcus mutans</i> and <i>412c</i>-deficient mutant strains

WANG Peng; FAN Zixuan; QIAO Li; DAI Yunjie; YE Zhaoyang; LIANG Yan

doi:10.7619/jcmp.20221342

Journal of Clinical Medicine in Practice > 2022 > 26(20): 70-77. > DOI: 10.7619/jcmp.20221342

WANG Peng, FAN Zixuan, QIAO Li, DAI Yunjie, YE Zhaoyang, LIANG Yan. Preliminary analysis in functions of clinical Streptococcus mutans and 412c-deficient mutant strains[J]. Journal of Clinical Medicine in Practice, 2022, 26(20): 70-77. DOI: 10.7619/jcmp.20221342

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Preliminary analysis in functions of clinical Streptococcus mutans and 412c-deficient mutant strains

1.
Department of Dentistry and Endodontics, School of Stomatology Affiliated to Guizhou Medical University, Guiyang, Guizhou, 550004
2.
School of Public Health of Guizhou Medical University, Guiyang, Guizhou, 550004
3.
Research Center of Clinical Medicine, Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, 550004

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Received Date: April 24, 2022
Available Online: November 03, 2022

Graphical Abstract

Abstract

Abstract

Objective
To isolate, purify and identify clinical Streptococcus mutans (S.mutans) strains, construct their 412c-deficient mutant strains, and investigate on their proliferation and cariogenicity.
Methods
Dental plaque samples were randomly collected from healthy adults without caries, were anaerobically cultivated on saliva or S.mutans bacitracin AGAR (MSA) medium and anaerobion were screened out. The isolated S.mutans strains were identified by colony morphology, Gram staining, biochemical test and polymerase chain reaction (PCR). The linearized recombinant plasmids were transformed into clinical S.mutans strains for homologous recombination to obtain 412c (hit) gene defect strains. The growth rate and acid resistance in the brain heart immersion (BHI) medium with different pH values. In BHI medium containing 3% sucrose, the acid production capacity of the two kinds of strains was compared.
Results
The morphology of the dental enamel surface plaque of healthy adults without caries growing in MSA medium was dark blue hemispheric granular protrusions with rough surface and uneven edges. Using Gram staining, there were many long and short chained coccus-shaped bluish violet bacterium under an oil-microscopy. The isolated S.mutans was identified as "an equivalent S.mutans UA159" by biochemical test and PCR-Sanger sequencing of 412c gene segment. The hit-UP-pFW5-Down recombinant plasmids were transformed into the genetic competence of the isolated S. mutans strains. The 412c-deficient mutant strains were screened out. Similar to S.mutans ATCC 25175, the growth rate of the 412c-deficient mutant strains increased greatly compared with their parental S.mutans strains.
Conclusion
The clinical isolate of S.mutans was "equivalent S.mutans UA159" strain, in which the 412c gene is growth-phase regulated, 412c-deficient S.mutans strains have significantly higher growth rates than its parental strain and have more cariogenic potential.
- Streptococcus mutans,
- 412c gene,
- growth cycle regulation,
- acidogenicity,
- resistance to acid,
- cariogenicity

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References (20)

References

[1]	CHEN D R, LIN H C. Research updates: cariogenic mechanism of Streptococcus mutans[J]. Journal of Sichuan University: Medical Science Edition, 2022, 53(2): 208-213.
[2]	ABRANCHES J, ZENG L, KAJFASZ J K, et al. Biology of oral Streptococci[J]. Microbiol Spectr, 2018, 6(5): 10.
[3]	ZHANG Q, MA Q Z, WANG Y, et al. Molecular mechanisms of inhibiting glucosyltransferases for biofilm formation in Streptococcus mutans[J]. Int J Oral Sci, 2021, 13(1): 30. doi: 10.1038/s41368-021-00137-1
[4]	MA Q Z, PAN Y Y, CHEN Y, et al. Acetylation of glucosyltransferases regulates Streptococcus mutans biofilm formation and virulence[J]. PLoS Pathog, 2021, 17(12): e1010134. doi: 10.1371/journal.ppat.1010134
[5]	FAUSTOVA M O, ANANIEVA M M, BASARAB Y O, et al. Bacterial factors of cariogenicity (literature review)[J]. Wiad Lek, 2018, 71(2): 378-382.
[6]	AJDIC D, MCSHAN W M, MCLAUGHLIN R E, et al. Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen[J]. PNAS, 2002, 99(22): 14434-14439. doi: 10.1073/pnas.172501299
[7]	YE Z Y, HOU Q M, LI L F, et al. Crystallization and preliminary X-ray crystallographic analysis of SMU. 412c protein from the caries pathogen Streptococcus mutans[J]. Acta Crystallogr Sect F Struct Biol Cryst Commun, 2009, 65(4): 392-394. doi: 10.1107/S1744309109009464
[8]	赖扬帆, 王鹏, 乔里, 等. 变异链球菌hit基因缺陷菌株的构建[J]. 口腔疾病防治, 2021, 29(12): 801-808. https://www.cnki.com.cn/Article/CJFDTOTAL-GDYB202112002.htm
[9]	LI S S, WU C Z, QIAO X H, et al. Advances on mechanism and treatment of salivary gland in radiation injury[J]. West China J Stomatol, 2021, 39(1): 99-104.
[10]	BACHTIAR E W, BACHTIAR B M. Relationship between Candida albicans and Streptococcus mutans in early childhood caries, evaluated by quantitative PCR[J]. F1000Res, 2018, 7: 1645. doi: 10.12688/f1000research.16275.2
[11]	BEDOYA-CORREA C M, RINCÓN RODRÍGUEZ R J, PARADA-SANCHEZ M T. Genomic and phenotypic diversity of Streptococcus mutans[J]. J Oral Biosci, 2019, 61(1): 22-31. doi: 10.1016/j.job.2018.11.001
[12]	KRAKOWIAK A, KOCON-REBOWSKA B, DOLOT R, et al. New interactions between tumor suppressor Fhit protein and a nonhydrolyzable analog of its AP4 A substrate[J]. FEBS Lett, 2017, 591(3): 548-559. doi: 10.1002/1873-3468.12560
[13]	COYKENDALL A L, LIZOTTE P A. Streptococcus mutans isolates identified by biochemical tests and DNA base contents[J]. Arch Oral Biol, 1978, 23(5): 427-428. doi: 10.1016/0003-9969(78)90104-8
[14]	CAI J N, CHOI H M, JEON J G. Relationship between sucrose concentration and bacteria proportion in a multi species biofilm: short title: Sucrose challenges to a multi species biofilm[J]. J Oral Microbiol, 2021, 13(1): 1910443. doi: 10.1080/20002297.2021.1910443
[15]	COSTA OLIVEIRA B E, RICOMINI FILHO A P, BURNE R A, et al. The route of sucrose utilization by Streptococcus mutans affects intracellular polysaccharide metabolism[J]. Front Microbiol, 2021, 12: 636684.
[16]	CAI J N, JUNG J E, LEE M H, et al. Sucrose challenges to Streptococcus mutans biofilms and the curve fitting for the biofilm changes[J]. FEMS Microbiol Ecol, 2018, 94(7): fiy091.
[17]	纪莹, 吴婷婷, 王欣, 等. 婴幼儿龋病口腔微生物群失调与唾液免疫生物标志物的关系研究[J]. 实用临床医药杂志, 2021, 25(9): 41-45. doi: 10.7619/jcmp.20201977
[18]	BAKER J L, SAPUTO S, FAUSTOFERRI R C, et al. Streptococcus mutans SpxA2 relays the signal of cell envelope stress from LiaR to effectors that maintain cell wall and membrane homeostasis[J]. Mol Oral Microbiol, 2020, 35(3): 118-128.
[19]	SEKIYA M, IZUMISAWA S, IWAMOTO-KIHARA A, et al. Proton-pumping F-ATPase plays an important role in Streptococcus mutans under acidic conditions[J]. Arch Biochem Biophys, 2019, 666: 46-51.
[20]	SHANMUGAM K, SARVESWARI H B, UDAYASHANKAR A, et al. Guardian genes ensuring subsistence of oral Streptococcus mutans[J]. Crit Rev Microbiol, 2020, 46(4): 475-491.

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Preliminary analysis in functions of clinical Streptococcus mutans and 412c-deficient mutant strains

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