Current Microbiology

, Volume 75, Issue 1, pp 11–19 | Cite as

Antimicrobial Mechanism of Oleanolic and Ursolic Acids on Streptococcus mutans UA159

  • Soon-Nang Park
  • Yun Kyong Lim
  • Mi-Hwa Choi
  • Eugene Cho
  • Iel Soo Bang
  • Jung Min Kim
  • Sug-Joon Ahn
  • Joong-Ki Kook


Triterpenoid saponin derivatives oleanolic acid (OA) and ursolic acid (UA), but not betulinic acid (BA), were previously found to have strong antimicrobial activity against Streptococcus mutans. OA and UA inhibited the transcription of genes related to peptidoglycan biosynthesis, thereby preventing bacterial growth. However, it is not clear whether this is the only pathway involved in the antimicrobial activity of these compounds against S. mutans. Therefore, we used quantitative real-time PCR (qPCR) and microarray analyses to examine the expression of genes related to essential metabolic pathways in S. mutans UA159 following incubation with OA, UA, or BA. An oligonucleotide array consisting of 5363 probes was designed to survey 1928 of the 1963 genes in the genome of S. mutans UA159. Genes that showed >2-fold changes in expression in response to the treatment conditions were annotated, and selected target genes involved in central metabolism were analyzed by qPCR. Microarray analysis confirmed that the gene expression patterns of the OA- and UA-treated cells differed from that of the BA-treated culture, indicating differences in the antimicrobial mechanism. In particular, the expression of pfk and pykF, coding for glycolysis regulatory proteins phosphofructokinase and pyruvate kinase, respectively, were significantly decreased in the OA and UA groups (P < 0.05), as were genes involved in fatty acid and amino acid synthesis. In addition, the microarray analysis confirmed previous qPCR results showing that peptidoglycan synthesis is down-regulated in the OA- and UA-treated groups. OA and UA also appear to decrease the generation of organic acids by S. mutans UA159, which would have an anticaries effect. Overall, these findings suggest that OA and UA affect multiple genes involved in the central metabolism of S. mutans, with inhibition of glycolysis, fatty acid synthesis, amino acid synthesis, and peptidoglycan synthesis, all contributing to their antimicrobial activity.



S. mutans UA 159 was generously provided by Dr. Robert A. Burne, Department of Oral Biology, College of Dentistry, University of Florida. This study was supported by a grant from the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare and Family Affairs, Republic of Korea (A091074) and, in part, by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning (2017M3A9B8065844).

Compliance with Ethical Standards

Conflict of interest

The authors have no conflict of interest to declare.

Supplementary material

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  1. 1.
    Ajdić D, McShan WM, McLaughlin RE, Savić G, Chang J, Carson MB, Primeaux C, Tian R, Kenton S, Jia H, Lin S, Qian Y, Li S, Zhu H, Najar F, Lai H, White J, Roe BA, Ferretti JJ (2002) Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc Natl Acad Sci USA 99:14434–14439. doi: 10.1073/pnas.172501299 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Banas JA (2004) Virulence properties of Streptococcus mutans. Front Biosci 9:1267–1277. doi: 10.2741/1305 CrossRefPubMedGoogle Scholar
  3. 3.
    Banno N, Akihisa T, Tokuda H, Yasukawa K, Higashihara H, Ukiya M, Watanabe K, Kimura Y, Hasegawa J, Nishino H (2004) Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor-promoting effects. Biosci Biotechnol Biochem 68:85–90. doi: 10.1271/bbb.68.85 CrossRefPubMedGoogle Scholar
  4. 4.
    Dashper SG, Reynolds EC (1992) pH regulation by Streptococcus mutans. J Dent Res 71:1159–1165CrossRefPubMedGoogle Scholar
  5. 5.
    Fontanay S, Grare M, Mayer J, Finance C, Duval RE (2008) Ursolic, oleanolic and betulinic acids: antibacterial spectra and selectivity indexes. J Ethnopharmacol 120:272–276. doi: 10.1016/j.jep.2008.09.001 CrossRefPubMedGoogle Scholar
  6. 6.
    Fozo EM, Quivey RG Jr (2004) The fabM gene product of Streptococcus mutans is responsible for the synthesis of monounsaturated fatty acids and is necessary for survival at low pH. J Bacteriol 186:4152–4158. doi: 10.1128/JB.186.13.4152-4158.2004 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Horiuchi K, Shiota S, Hatano T, Yoshida T, Kuroda T, Tsuchiya T (2007) Antimicrobial activity of oleanolic acid from Salvia officinalis and related compounds on vancomycin-resistant enterococci (VRE). Biol Pharm Bull 30:1147–1149. doi: 10.1248/bpb.30.1147 CrossRefPubMedGoogle Scholar
  8. 8.
    Kim MJ, Kim CS, Ha WH, Kim BH, Lim YK, Park SN, Cho YJ, Kim M, Ko JH, Kwon SS, Ko YM, Kook JK (2010) Antimicrobial effects of oleanolic acid against Streptococcus mutans and Streptococcus sobrinus isolated from a Korean population. Int J Oral Biol 35:191–195Google Scholar
  9. 9.
    Kim MJ, Kim CS, Park JY, Lim YK, Park SN, Ahn SJ, Jin DC, Kim TH, Kook JK (2011) Antimicrobial effects of ursolic acid against mutans streptococci isolated from Koreans. Int J Oral Biol 36:7–11Google Scholar
  10. 10.
    Liu J (1995) Pharmacology of oleanolic acid and ursolic acid. J Ethnopharmacol 49:57–68CrossRefPubMedGoogle Scholar
  11. 11.
    Ovesná Z, Vachálková A, Horváthová K, Tóthová D (2004) Pentacyclic triterpenoic acids: new chemoprotective compounds. Neoplasma 51:327–333PubMedGoogle Scholar
  12. 12.
    Park SN, Ahn SJ, Kook JK (2015) Oleanolic acid and ursolic acid inhibit peptidoglycan biosynthesis in Streptococcus mutans UA159. Braz J Microbiol 46:613–617. doi: 10.1590/S1517-838246246220130209 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Torres-Santos EC, Lopes D, Oliveira RR, Carauta JP, Falcao CA, Kaplan MA, Rossi-Bergmann B (2004) Antileishmanial activity of isolated triterpenoids from Pourouma guianensis. Phytomedicine 11:114–120. doi: 10.1078/0944-7113-00381 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Soon-Nang Park
    • 1
  • Yun Kyong Lim
    • 1
  • Mi-Hwa Choi
    • 1
  • Eugene Cho
    • 1
  • Iel Soo Bang
    • 2
    • 3
  • Jung Min Kim
    • 4
  • Sug-Joon Ahn
    • 5
  • Joong-Ki Kook
    • 1
    • 3
  1. 1.Korean Collection for Oral Microbiology and Department of Oral Biochemistry, School of DentistryChosun UniversityGwangjuRepublic of Korea
  2. 2.Department of Oral Microbiology, School of DentistryChosun UniversityGwangjuRepublic of Korea
  3. 3.Oral Biology Research InstituteChosun UniversityGwangjuRepublic of Korea
  4. 4.Genoplan Korea, Inc. and NAR Center, Inc.SeoulRepublic of Korea
  5. 5.Dental Research Institute and Department of Orthodontics, School of DentistrySeoul National UniversitySeoulRepublic of Korea

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