• Journal of the korean academy of Pediatric Dentistry
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• v.47 no.4
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• pp.397-405
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• 2020

The aim of this study is to compare cariogenic characteristics of fluoride-sensitive Streptococcus mutans [fluoride-sensitive (FS) S. mutans ] and fluoride-resistant Streptococcus mutans [fluoride-resistant (FR) S. mutans] in the presence of sucrose, and to evaluate its effect on cariogenic biofilm formation. S. mutans ATCC 25175 was continuously cultured in trypticase soy broth (TSB) containing NaF (70 ppm) for 40 days to generate FR S. mutans. FS and FR S. mutans were inoculated in TSB with or without 2% sucrose, and optical density and pH were measured every hour. An oral biofilm was formed using saliva bacteria and analyzed through confocal laser scanning microscopy and CFU count. Finally, the expression of glucosyltransferases genes of both S. mutans was investigated through RT-PCR. FR S. mutans exhibited slower growth and lower acidogenicity in the presence of sucrose compared to FS S. mutans . Both cariogenic and single species biofilm formation was lower in the presence of FR S. mutans, along with reduced number of bacteria. FR S. mutans showed significantly low levels of gtfB, gtfC, and gtfD expression compared to FS S. mutans . On the basis of results, FR S. mutans may be less virulent in the induction of dental caries.

• Journal of Life Science
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• v.29 no.1
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• pp.90-96
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• 2019

The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.1
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• pp.129-141
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• 2001

When oral microorganisms were sampled from occlusal surfaces of caries and non-caries teeth, $3.43\times10^5$ CFU and $3.47\times10^3$ CFU of bacteria were counted on MSB agar plates, respectively. All the 20 colonies isolated from a caries surface were Streptococcus mutans but, only two of 20 colonies were identified as Streptococcus mutans by API test. S. mutans SM1 from caries tooth and S. mutans SM2 from non-caries tooth showed the same results except for $\alpha-galactosidase$ activity on sugar fermentation tests and biochemical tests. For the bacterial replication, both SM1 and SM2 were actively multiplicated at pH 5.5. And the viability of SM1 was high at 20% of sucrose, while that of SM2 was high at 5% of sucrose in the media. SM1 actively replicated at 16mM of $CaCl_2$, 160mM of KCl, and 6.4mM of $MgCl_2$, and the replication of SM2 was increased at 16mM of $CaCl_2$, 40mM of KCl, 6.4mM of $MgCl_2$. At 1mM of sodium bicarbonate and sodium phosphate, both bacteria were actively multiplicated. SM1 and SM2 were actively replicated at 1mM and 10mM of Tris, respectively. For potassium phosphate buffer, SM1 grew well proportionally to the concentration up to 100mM, while the growth of SM2 were inhibited by the increase of concentration. The 4.6 kb of gtf gene was amplified with a pair of primer, gtfB-F961 and gtfC-R5574 by polymerase chain reaction from the chromosomal DNA of SM1 and SM2. When 4.6kb bands were eluted from gel and were treated with restriction enzyme, EcoR I produced the same RFLP like 0.8kb and 3.8kb of DNA fragments for S. mutans GS-5, SM1 and SM2. By Hind III, the PCR products weren't digested for S. mutans GS-5 and SM1, but 3 fragments such as 2.4kb, 1.8kb and 400bp were examined for SM2. These results indicated the difference between gtf genes of SM1 and SM2. BamH I treatment showed 4 fragments for SM1 and SM2, while the 3 fragments for S. mutans GS-5. The PCR products were not digested by Kpn I, Sma I, Xho I and Pst I.

• Journal of Applied Biological Chemistry
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• v.55 no.3
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• pp.173-178
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• 2012

Sucrose-glucan glucosyltransferase (Gtf) is an important enzyme involved in the cavity formation process where insoluble glucan is synthesized. In this study, we purified Gtf from Streptcoccus mutans Ingbritt through ammonium sulfate precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex column chromatographies. A 13-fold of purification was achieved with a total yield of 6.3%. The apparent molecular mass of the enzyme was determined to be 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal pH and temperature were established to be 6.0 and $40^{\circ}C$, respectively. The enzyme activity could be inhibited to 22-59% by 1 mM $Hg^{2+}$, $Cu^{2+}$ and $Al^{3+}$, and to 68% by 1 mM EDTA. It was also inhibited 40% by 2 mM xylitol and 35-45% by 0.05% soluble chitosan, glycol chitosan, and glycol chitin. This is the first report to reveal the inhibition effect of chitin derivatives on Gtf activity, which may be further applicable to develop gargles to overcome cavity.

• Proceedings of the KAIS Fall Conference
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• 2009.12a
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• pp.498-500
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• 2009

본 연구에서는 한국인 아동의 우식치아로부터 분리한 S. mutans K7으로부터 HtrA gene을 동정하고 HtrA expression이 산성환경하에서 S. mutans의 생육에 미치는 영향을 알아보았다. S. mutans K7의 HtrA mutant strain은 산성환경에서 parental strain과 비교하였을 때, 생육에 있어서 상당한 차이를 나타내었다. 또한 Biofilm formation에 관여하는 GtfB, 와 GtfC의 발현량도 현저히 줄어들었다. 그리고 HtrA mutant strain에 HtrA gene을 삽입하여 HtrA의 발현량을 회복하였을 경우에는 acid stress하에서 control과 같은 nomal growth phenotype을 회복하였다. 이러한 결과들은 S. mutans K7에서 HtrA가 acid stress동안에 중요한 역할을 담당함을 제시하고 있다.

• Journal of dental hygiene science
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• v.21 no.2
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• pp.119-126
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• 2021

Background: Dental caries is mainly composed of various cellular components and is deposited around the tooth surface and gums, causing a number of periodontal diseases. Streptococcus mutans is commonly found in the human oral cavity and is a significant contributor to tooth decay. The use of antibacterial ingredients in oral hygiene products has demonstrated usefulness in the management of dental caries. This study investigated the anticaries effect of the ethanol extract of Terminalia chebula (EETC) against S. mutans and their cytotoxicity to gingival epithelial cells. Methods: The EETC was prepared from T. chebula fruit using ethanol extraction. Disk diffusion, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and colony forming unit (CFU) were analyzed to investigate the antimicrobial activity of the EETC. Glucan formation was measured using the filtrate of the bacterial culture medium and sucrose. Gene expression was analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Cytotoxicity was analyzed via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Results: The antibacterial activity of the EETC was explored using disc diffusion and CFU measurements. The MIC and MBC of the EETC were 10 and 20 ㎍/ml, respectively. EETC treatment decreased insoluble glucan formation by S. mutans enzymes and also resulted in reduced glycosyltransferase B (gtf B), gtf C, gtf D, and fructosyltransferase (ftf), expressions on RT-PCR. In addition, at effective antibacterial concentrations, EETC treatment was not cytotoxic to gingival epithelial cells. Conclusion: These results demonstrate that the EETC is an effective anticaries ingredient with low cytotoxicity to gingival epithelial cells. The EETC may be useful in antibacterial oral hygiene products for the management of dental caries.

• Molecules and Cells
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• v.38 no.5
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• pp.402-408
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• 2015

Protein O-GlcNAcylation, dictated by cellular UDP-N-acetylglucosamine (UDP-GlcNAc) levels, plays a crucial role in posttranslational modifications. The enzyme GlcNAc kinase (NAGK, E.C. 2.7.1.59) catalyzes the formation of GlcNAc-6-phosphate, which is a major substrate for the biosynthesis of UDP-GlcNAc. Recent studies have revealed the expression of NAGK in different types of cells especially in neuronal dendrites. Here, by immunocytochemistry (ICC) and immunonucleochemistry (INC) of cultured rat hippocampal neurons, HEK293T and GT1-7 cells, we have showed that NAGK immuno-reactive punctae being present in the nucleoplasm colocalized with small nuclear ribonucleoprotein-associated protein N (snRNPN) and p54NRB, which are speckle and paraspeckle markers, respectively. Furthermore, NAGK IR cluster was also found to be colocalized with GTF2H5 (general transcription factor IIH, polypeptide 5) immuno reactive punctae. In addition, relative localization to the ring of nuclear lamin matrix and to GlcNAc, which is highly enriched in nuclear pore complexes, showed that NAGK surrounds the nucleus at the cytoplasmic face of the nuclear outer membrane. By in situ proximity ligation assay (PLA) we confirmed the colocalization of NAGK with snRNPN in the nucleus and in dendrites, while we also verified the interactions of NAGK with p54NRB, and with GTF2H5 in the nucleus. These associations between NAGK with speckle, paraspeckle and general transcription factor suggest its regulatory roles in gene expression.

• Journal of Microbiology and Biotechnology
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• v.2 no.4
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• pp.243-247
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• 1992

The thr operon of Escherichia coli TF427, an $\alpha$-amino-$\beta$-hydroxyvaleric acid (AHV)-resistant threonine overproducer, was cloned in a pBluescriptII $KS^＋$ plasmid by complementation of E. coli mutants. All clones contained a common 8.8 kb HindIII-generated DNA fragment and complemented the thrA, thrB, and thrC mutants by showing that these clones contained the whole thr operon. This thr operon was subcloned in the plasmid vectors pBR322, pUC18, and pECCG117, an E. coli/Corynebacterium glutamicum shuttle vector, to form recombinant plasmids pBTF11, pUTF25 and pGTF18, respectively. The subcloned thr operon was shown to be present in a 6.0 kb insert. A transformant of E. coli TF125 with pBTF11 showed an 8~11 fold higher aspartokinase I activity, and 15~20 fold higher L-threonine production than TF125, an AHV-sensitive methionine auxotroph. Also, it was found that the aspartokinase I activity of E. coli TF125 harboring pBTF11 was not inhibited by threonine and its synthesis was not repressed by threonine plus isoleucine.

• Journal of Ginseng Research
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• v.39 no.1
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• pp.54-60
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• 2015

Background: The present study was designed to prepare and find the optimum active preparation or fraction from Korea Red Ginseng inhibiting matrix metalloproteinase-13 (MMP-13) expression, because MMP-13 is a pivotal enzyme to degrade the collagen matrix of the joint cartilage. Methods: From total red ginseng ethanol extract, n-BuOH fraction (total ginsenoside-enriched fraction), ginsenoside diol-type-enriched fraction (GDF), and ginsenoside triol-type-enriched fraction (GTF) were prepared, and ginsenoside diol type-/F4-enriched fraction (GDF/F4) was obtained from Panax ginseng leaf extract. Results: The n-BuOH fraction, GDF, and GDF/F4 clearly inhibited MMP-13 expression compared to interleukin-$1{\beta}$-treated SW1353 cells (human chondrosarcoma), whereas the total extract and ginsenoside diol-type-enriched fraction did not. In particular, GDF/F4, the most effective inhibitor, blocked the activation of p38 mitogen-activated protein kinase (p38 MAPK), c-Jun-activated protein kinase (JNK), and signal transducer and activator of transcription-1/2 (STAT-1/2) among the signal transcription pathways involved. Further, GDF/F4 also inhibited the glycosaminoglycan release from interleukin-$1{\alpha}$-treated rabbit cartilage culture (30.6% inhibition at $30{\mu}g/mL$). Conclusion: Some preparations from Korean Red Ginseng and ginseng leaves, particularly GDF/F4, may possess the protective activity against cartilage degradation in joint disorders, and may have potential as new therapeutic agents.