Skip to main content
SpringerLink
Log in

Identification and determination of antibacterial constituents in residue discharged from garlic-processing plant

  • Original Paper
  • Published:
European Food Research and Technology Aims and scope Submit manuscript

Abstract

Activity-guided fractionation based on in vitro assays was used to identify antibacterial components in the residue wasted in the manufacturing process of aged garlic extract (AGE). Antimicrobial activity of the hexane extract of aged garlic-derived residue (AGR) was evaluated against clinically isolated Gram-positive and -negative bacteria including levofloxacin (LVX)-resistant strains. Among antimicrobial fractions separated from the hexane extract of AGR, several polysulfides, such as diallyl-tetrasulfide (DAS4), -pentasulfide (DAS5) and -hexasulfide (DAS6), were identified as active components, whose chemical structures were determined by means of LC–MS and NMR spectroscopy. We found that those sulfides exhibited antimicrobial activities against Gram-positive bacteria at minimum inhibitory concentrations (MICs) ranging from 32 to 80 μg/mL. Although these sulfides were less active than LVX against most of the bacteria examined, they were more potent against Staphylococcus aureus (S. aureus) including methicillin-resistant S. aureus (MRSA). Surprisingly, DAS5 significantly suppressed the biofilm formation of S. aureus by 60% at a concentration of 1.25 μg/mL, which is extremely lower than its MIC value. These results show that AGR contains several antimicrobial diallyl-polysulfides which have the potential to inhibit the growth and biofilm formation of bacteria and AGR may be a useful bioresource as a natural additive for antibiotic drugs or foods.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Verified Market Research (2018) Global plant extracts market by type, by application, by geography, size and forecast to 2025. Electron Rep (report ID: 5093)

  2. Wei Z, Yang Y, Yang R, Wang C (2012) Alkaline lignin extracted from furfural residues for pH-responsive pickering emulsions and their recyclable polymerization. Green Chem 14:3230–3236

    Article  CAS  Google Scholar 

  3. Akasaka H, Ueki K, Ueki A (2004) Effects of plant residue extract and cobalamin on growth and propionate production of Propionicimonas paludicola isolated from plant residue in irrigated rice field soil. Microbes environ 19(2):112–119

    Article  Google Scholar 

  4. Ubwa ST, Asemave K, Oshido B, Idoko A (2013) Preparation of biogas from plants and animal waste. Int J Sci Technol 2(6):480–485

    Google Scholar 

  5. Truong NM, Phung TT, Do TK, Nguyen VQ, Pham TTH, Nguyen TQ, Yusuf A, Xinyan F, Truong MV, Tran DK, Tran DX (2017) Potential use of plant waste from the moth orchid (phalaenopsis sogo yukidian “V3”) as an antioxidant source. Foods. https://doi.org/10.3390/foods6100085

    Article  Google Scholar 

  6. Xiong F, Dai CH, Hou FR, Zhu PP, He RH, Ma HL (2018) Study on the ageing method and antioxidant activity of black garlic resides. Czech J Food Sci 36(1): 88–97. https://doi.org/10.17221/420/2016-CJFS

  7. Kallel F, Driss D, Chaari F, Ellouz SC (2014) Garlic (Allium sativum L.) husk waste as a potential source of phenolic compounds: Influence of extracting solvents on its antimicrobial and antioxidant properties. Ind Crops Prod 62:34–41. https://doi.org/10.1016/j.indcrop.2014.07.047

    Article  CAS  Google Scholar 

  8. Amagase H (2006) Clarifying the real bioactive constituents of garlic. J Nutr 136(Suppl 3):716–725

    Article  Google Scholar 

  9. Ryu K, Ide N, Matsuura H, Itakura Y (2001) Nα-(1-deoxy-d-fructos-1-yl)-l-arginine, an antioxidant compound identified in aged garlic extract. J Nutr 131(3):972S–976S

    Article  CAS  PubMed  Google Scholar 

  10. Matsutomo T, Kodera Y (2016) Development of an analytic method for sulfur compounds in aged garlic extract with the use of a postcolumn high performance liquid chromatography method with sulfur specific detection. J Nutr 146(2S):450S–455S

    Article  CAS  PubMed  Google Scholar 

  11. Ichikawa M, Ryu K, Yoshida J, Ide N, Yoshida S, Sasaoka T, Sumi S (2002) Antioxidant effects of tetrahydro-beta-carboline derivatives identified in aged garlic extract. BioFactors 16:57–62

    Article  CAS  PubMed  Google Scholar 

  12. Matsutomo T, Stark TD, Hofmann T (2018) Targeted screening and quantitative analyses of antioxidant compounds in aged garlic extract. Euro Food Res Technol 244(10):1803–1814

    Article  CAS  Google Scholar 

  13. Imai J, Ide N, Nagae S, Moriguchi T, Matsuura H, Itakura Y (1994) Antioxidant and radical scavenging effects of aged garlic extract and its constituents. Planta Med 60(5):417–420

    Article  CAS  PubMed  Google Scholar 

  14. Nakagawa S, Kasuga S, Matsuura H (1989) Prevention of liver damage by aged garlic extract and its components in mice. Phytother Res 3(2):50–53

    Article  CAS  Google Scholar 

  15. Kyo E, Uda N, Kasuga S, Itakura Y (1079S) Immunomodulatory effects of aged garlic extract. J Nutr 131:1075S–1079S

    Article  CAS  PubMed  Google Scholar 

  16. Ushijima M, Takashima M, Kunimura K, Kodera Y, Morihara N, Tamura K (2018) Effects of S-1propenylcysteine, a sulfur compound in aged garlic extract, on blood pressure and peripheral circulation in spontaneously hypertensive rats. J Pharm Pharmacol 70(4):559–565

    Article  CAS  PubMed  Google Scholar 

  17. Koch HP, Lawson LD (1996) Garlic: the science and therapeutic application of Allium sativum L. and related species. Williams and Wilkins, Baltimore

    Google Scholar 

  18. Kim JW, Huh JE, Kyung SH, Kyung KH (2004) Antimicrobial activity of alk(en)yl sulfides found in essential oils of garlic and onion. Food Sci Biotechnol 13:235–239

    CAS  Google Scholar 

  19. Coralie OMM, David L, Catrin W, David W, Gareth E, Robert AS, Joanne C (2011) Effect of garlic and allium-derived products on growth and metabolism of Spironucleus vortens. Exp Parasitol 127(2):490–499

    Article  Google Scholar 

  20. Zwergal A (1952) Beitrag zur Kenntnis der Inhaltsstoffe des Knoblauchs Allium sativum L. Chem Abst 47:3224

    Google Scholar 

  21. Tsao SM, Yin M (2001) In vitro activity of garlic oil and four diallyl sulphides against antibiotic-resistant Pseudomonas aeruginosa and Klebsiella pneumoniae. J Antimicrob Chemother 47:665–670

    Article  CAS  PubMed  Google Scholar 

  22. Tsao SM, Yin M (2001) In-vitro antimicrobial activity of four diallyl sulphides occurring naturally in garlic and Chinese leek oils. J Med Microbiol 50:646–649

    Article  CAS  PubMed  Google Scholar 

  23. O’gara EA, Hill DJ, Maslin DJ (2000) Activities of garlic oil, garlic powder, and their diallyl constituents against Helicobacter pylori. Appl Environ Microbiol 66(5):2269–2273

    Article  PubMed  PubMed Central  Google Scholar 

  24. Oosthuizen C, Arbach M, Meyer D, Hamilton H, Lall C (2017) Diallyl polysulfides from allium sativum as immunomodulators, hepatoprotectors, and antimycobacterial agents. J Med Food 20(7):685–690. https://doi.org/10.1089/jmf.2016.0137

    Article  CAS  PubMed  Google Scholar 

  25. Bjarnsholt T, Jensen PO, Rasmussen TB, Christophersen L, Calum H (2005) Garlic blocks quorum sensing and promotes rapid clearing of pulmonary Pseudomonas aeruginosa infections. Microbiology 151:3873–3880

    Article  CAS  PubMed  Google Scholar 

  26. Christensen LD, van Gennip M, Jakobsen TH, Alhede M, Hougen HP (2012) Synergistic antibacterial efficacy of early combination treatment with tobramycin and quorum-sensing inhibitors against Pseudomonas aeruginosa in an intraperitoneal foreign-body infection mouse model. J Antimicrob Chemother 67:1198–1206

    Article  CAS  PubMed  Google Scholar 

  27. Jakobsen TH, van Gennip M, Phipps RK, Shanmugham MS, Christensen LD (2012) Ajoene, a sulfur rich molecule from garlic, inhibits genes controlled by quorum sensing. Antimicrob Agents Chemother 56(5):2314–2325

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. López D, Vlamakis H, Kolter R (2010) Biofilm. Cold Spring Harb Perspect Biol. https://doi.org/10.1101/cshperspect.a000398

    Article  PubMed  PubMed Central  Google Scholar 

  29. Muhsin J, Ufaq T, Tahir H, Saadia A (2015) Bacterial biofilm: its composition, formation and role in human infections. J Microbiol Biotechnol 4:1–14

    Google Scholar 

  30. Bassler BL, Small talk (2002) Cell-to-cell communication in bacteria. Cell 109(4):421–424

    Article  CAS  PubMed  Google Scholar 

  31. Nadell CD, Xavier JB, Levin SA, Foster KR (2008) The evolution of quorum sensing in bacterial biofilms. PLoS Biol 6(1):e14

    Article  PubMed  PubMed Central  Google Scholar 

  32. Xueqing W, Regiane R, Johanna FG (2015) Analyzing the antibacterial effects of food ingredienta: model experiments with allicin and garlic extracts on biofilm formation and viability of Staphylococcus epidermidis. Food Sci Nutr 3(2):158–168

    Article  Google Scholar 

  33. Li WR, Ma YK, Shi QS, Xie XB, Sun TL, Peng H, Huang XM (2018) Diallyl disulfide from garlic oil inhibits Pseudomonas aeruginosa virulence factors by inactivating key quorum sensing genes. Appl Microbiol Biotechnol 102(17):7555–7564

    Article  CAS  PubMed  Google Scholar 

  34. Ried K, Travica N, Saji A (2018) The effect of kyolic aged garlic extract on gut microbiota, inflammation, and cardiovascular markers in hypertensives: the GarGIC trial. Front Nutr 11(5):122. https://doi.org/10.3389/fnut.2018.00122

    Article  CAS  Google Scholar 

  35. Ried K, Travica N, Saji A (2016) The effect of aged garlic extract on blood pressure and other cardiovascular risk factors in uncontrolled hypertensives: the AGE at Heart trial. Integr Blood Press Control 27(9):9–21. https://doi.org/10.2147/IBPC.S93335

    Article  Google Scholar 

  36. Freeman F, Kodera Y (1995) Garlic chemistry: Stability of S-(2-propyl) 2-propen-1-sulfinothioate (allicin) in blood, solvents, and stimulated physiological fluids. J Agric Food Chem 43:2332–2338

    Article  CAS  Google Scholar 

  37. Baker A, Graz M, Saunders R, Evans GJS, Kaul S, Wirth T (2013) Flow synthesis of symmetrical di- and trisulfides using phase-transfer catalysis. J Flow Chem 3(4):118–121

    Article  Google Scholar 

  38. Müller A, Eller J, Albrecht F, Prochnow P, Kuhlmann K, Bandow JE, Slusarenko AJ, Leichert LI (2016) Allicin induces thiol stress in bacteria through S-allylmercapto modification of protein cysteines. J Biol Chem 291(22):11477–11490

    Article  PubMed  PubMed Central  Google Scholar 

  39. Reardon-Robinson ME, Osipiuk J, Chang C, Wu C, Jooya N, Joachimiak A, Das A, Ton-That H (2015) A disulfide bond-forming machine is linked to the sortase-mediated pilus assembly pathway in the Gram-positive bacterium Actinomyces oris. J Biol Chem 290:21393–21405

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Reardon-Robinson ME, Osipiuk J, Jooya N, Chang C, Joachimiak A, Das A, Ton-That H (2015) A thiol-disulfide oxidoreductase of the Gram-positive pathogen Corynebacterium diphtheriae is essential for viability, pilus assembly, toxin production and virulence. Mol Microbiol. https://doi.org/10.1111/mmi.13172

    Article  PubMed  PubMed Central  Google Scholar 

  41. Cady NC, McKean KA, Behnke J, Kubec R, Mosier AP, Kasper SH, Burz DS, Musah RA (2012) Inhibition of biofilm formation, quorum sensing and infection in pseudomonas aeruginosa by natural products-inspired organosulfur compounds. PLoS ONE. https://doi.org/10.1371/journal.pone.0038492

    Article  PubMed  PubMed Central  Google Scholar 

  42. Saquib SA, AlQahtani NA, Ahmad I, Kader MA, Al Shahrani SS, Asiri EA (2019) Evaluation and comparison of antibacterial efficacy of herbal extracts in combination with antibiotics on periodontal pathobionts: an in vitro microbiological study. Antibiotics (Basel). https://doi.org/10.3390/antibiotics8030089

    Article  Google Scholar 

  43. Williams CF, Vacca AR, Dunham L, Lloyd D, Coogan MP, Evans G, Graz M, Cable J (2016) The redox-active drug metronidazole and thiol-depleting garlic compounds act synergistically in the protist parasite Spironucleus vortens. Mol Biochem Parasitol 206(1–2):20–28. https://doi.org/10.1016/j.molbiopara.2016.03.001

    Article  CAS  PubMed  Google Scholar 

  44. Plata-Rueda A, Martínez LC, Santos MHD, Fernandes FL, Wilcken CF, Soares MA, Serrão JE, Zanuncio JC (2017) Insecticidal activity of garlic essential oil and their constituents against the mealworm beetle, Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae). Sci Rep 20(7):46406. https://doi.org/10.1038/srep46406

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors deeply thank Dr. Takami Oka of Wakunaga Pharmaceutical Co., Ltd, for his helpful advice, encouragement and critical reading of the manuscript. The authors are grateful to Keiichi Hiramatsu, PhD, at JuntendoUniversity (Tokyo, Japan) and Yoshihito Otsuka, PhD, at Kameda Medical Center (Chiba, Japan) for supplying bacterial isolates.

Author information

Authors and Affiliations

  1. Central Research Institute, Wakunaga Pharmaceutical Co., Ltd., 1624 Shimokotachi, Koda–cho, Akitakata, Hiroshima, 739-1195, Japan

    Masato Nakamoto, Kayo Kunimura, Hirotaka Amano & Junichiro Wakamatsu

  2. Drug Discovery Laboratory, Wakunaga Pharmaceutical Co., Ltd., 1624 Shimokotachi, Koda–cho, Akitakata, Hiroshima, 739-1195, Japan

    Kensuke Ohishi

Authors
  1. Masato Nakamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kensuke Ohishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kayo Kunimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hirotaka Amano
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Junichiro Wakamatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Masato Nakamoto.

Ethics declarations

Conflict of interest

The authors declare no conflict of interests.

Compliance with Ethics requirements

Neither human nor animal rights were violated in this work.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

217_2020_3460_MOESM1_ESM.doc

Chromatograms obtained by LC-MS and HPLC, MIC values of fractions against Gram-Negative strains, and data of safety studies using DAS1-6 are shown in supplementary material (DOCX 136 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nakamoto, M., Ohishi, K., Kunimura, K. et al. Identification and determination of antibacterial constituents in residue discharged from garlic-processing plant. Eur Food Res Technol 246, 1041–1049 (2020). https://doi.org/10.1007/s00217-020-03460-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00217-020-03460-9

Keywords

Search

Navigation