Evaluation of Cocoa Bean Shell Antimicrobial Activity: A Tentative Assay Using a Metabolomic Approach for Active Compound Identification^{[ # ]}

Abstract

Cocoa bean shell is one of the main by-products of chocolate manufacturing and possesses several compounds with biofunctionalities. It can function as an antibacterial agent, and its action is mostly reported against Streptococcus mutans. However, only a few studies have investigated the cocoa bean shell compounds responsible for this activity. This study aimed to evaluate several extracts of cocoa bean shells from different geographical origins and cocoa varieties and estimate their antimicrobial properties against different fungal and bacterial strains by determining their minimal inhibitory concentration. The results demonstrated antimicrobial activity of cocoa bean shell against one of the tested strains, S. mutans. Cocoa bean shell extracts were further analysed via LC-HRMS for untargeted metabolomic analysis. LC-HRMS data were analysed (preprocessing and statistical analyses) using the Workflow4Metabolomics platform. The latter enabled us to identify possible compounds responsible for the detected antimicrobial activity by comparing the more and less active extracts. Active extracts were not the most abundant in polyphenols but contained higher concentrations of two metabolites. After tentative annotation of these metabolites, one of them was identified and confirmed to be 7-methylxanthine. When tested alone, 7-methylxanthine did not display antibacterial activity. However, a possible cocktail effect due to the synergistic activity of this molecule along with other compounds in the cocoa bean shell extracts cannot be neglected. In conclusion, cocoa bean shell could be a functional ingredient with benefits for human health as it exhibited antibacterial activity against S. mutans. However, the antimicrobial mechanisms still need to be confirmed.

Key words

cocoa bean shell - Theobroma cacao - Malvaceae - antimicrobial activity - Streptococcus mutans - metabolomics

^# Dedicated to Professor Arnold Vlietinck on the occasion of his 80th birthday.

Publikationsverlauf

Eingereicht: 29. Dezember 2020

Angenommen nach Revision: 23. April 2021

Artikel online veröffentlicht:
21. Mai 2021

• References

• 1 Rios J, Recio M. Medicinal plants and antimicrobial activity. J Ethnopharmacol 2005; 100: 80-84
• 2 Barbieri R, Coppo E, Marchese A, Daglia M, Sobarzo-Sanchez E, Nabavi SF, Nabavi SM. Phytochemicals for human disease: An update on plant-derived compounds antibacterial activity. Microbiol Res 2017; 196: 44-68
• 3 World Health Organization. Global antimicrobial resistance surveillance system (GLASS) report: early implementation 2017–2018. Accessed December 10, 2020 at: https://apps.who.int/iris/bitstream/handle/10665/279656/9789241515061-eng.pdf?ua=1
• 4 Daglia M. Polyphenols as antimicrobial agents. Curr Opin Biotechnol 2012; 23: 174-181
• 5 Oliveira Ribeiro S, Fontaine V, Mathieu V, Zhiri A, Baudoux D, Stévigny C, Souard F. Antibacterial and cytotoxic activities of ten commercially available essential oils. Antibiotics (Basel) 2020; 9: 717
• 6 Guil-Guerrero J, Ramos L, Moreno C, Zúñiga-Paredes J, Carlosama-Yepez M, Ruales P. Antimicrobial activity of plant-food by-products: A review focusing on the tropics. Livest Sci 2016; 189: 32-49
• 7 Widsten P, Cruz CD, Fletcher GC, Pajak MA, McGhie TK. Tannins and extracts of fruit byproducts: antibacterial activity against foodborne bacteria and antioxidant capacity. J Agric Food Chem 2014; 62: 11146-11156
• 8 Rojo-Poveda O, Barbosa-Pereira L, Zeppa G, Stévigny C. Cocoa bean shell – A by-product with nutritional properties and biofunctional potential. Nutrients 2020; 12: 1123
• 9 Hernández-Hernández C, Morales-Sillero A, Fernández-Prior MÁ, Fernández-Bolaños J, de la Paz Aguilera-Herrera M, Rodríguez-Gutiérrez G. Extra virgin olive oil jam enriched with cocoa bean husk extract rich in theobromine and phenols. LWT-Food Sci Technol 2019; 111: 278-283
• 10 Rojo-Poveda O, Barbosa-Pereira L, Mateus-Reguengo L, Bertolino M, Stévigny C, Zeppa G. Effects of particle size and extraction methods on cocoa bean shell functional beverage. Nutrients 2019; 11: 867
• 11 Cantele C, Rojo-Poveda O, Bertolino M, Ghirardello D, Cardenia V, Barbosa-Pereira L, Zeppa G. In vitro bioaccessibility and functional properties of phenolic pompounds from enriched beverages based on cocoa bean shell. Foods 2020; 9: 715
• 12 Nsor-Atindana J, Zhong F, Mothibe KJ, Bangoura ML, Lagnika C. Quantification of total polyphenolic content and antimicrobial activity of cocoa (Theobroma cacao L.) bean shells. Pak J Nutr 2012; 11: 574
• 13 Ooshima T, Osaka Y, Sasaki H, Osawa K, Yasuda H, Matsumura M, Sobue S, Matsumoto M. Caries inhibitory activity of cacao bean husk extract in in-vitro and animal experiments. Arch Oral Biol 2000; 45: 639-645
• 14 Osawa K, Miyazaki K, Shimura S, Okuda J, Matsumoto M, Ooshima T. Identification of cariostatic substances in the cacao bean husk: their anti-glucosyltransferase and antibacterial activities. J Dent Res 2001; 80: 2000-2004
• 15 Taguri T, Tanaka T, Kouno I. Antibacterial spectrum of plant polyphenols and extracts depending upon hydroxyphenyl structure. Biol Pharm Bull 2006; 29: 2226-2235
• 16 Adi-Dako O, Ofori-Kwakye K, Manso SF, Boakye-Gyasi ME, Sasu C, Pobee M. Physicochemical and antimicrobial properties of cocoa pod husk pectin intended as a versatile pharmaceutical excipient and nutraceutical. J Pharm (Cairo) 2016; 2016: 7608693
• 17 Santos R, Oliveira D, Sodré G, Gosmann G, Brendel M, Pungartnik C. Antimicrobial activity of fermented Theobroma cacao pod husk extract. Genet Mol Res 2014; 13: 7725-7735
• 18 Todorovic V, Milenkovic M, Vidovic B, Todorovic Z, Sobajic S. Correlation between antimicrobial, antioxidant activity, and polyphenols of alkalized/nonalkalized cocoa powders. J Food Sci 2017; 82: 1020-1027
• 19 Ho VTT, Zhao J, Fleet G. Yeasts are essential for cocoa bean fermentation. Int J Food Microbiol 2014; 174: 72-87
• 20 Collar C, Rosell CM, Muguerza B, Moulay L. Breadmaking performance and keeping behavior of cocoa-soluble fiber-enriched wheat breads. Food Sci Technol Int 2009; 15: 79-87
• 21 Rojo-Poveda O, Barbosa-Pereira L, Orden D, Stévigny C, Zeppa G, Bertolino M. Physical properties and consumer evaluation of cocoa bean shell-functionalized biscuits adapted for diabetic consumers by the replacement of sucrose with tagatose. Foods 2020; 9: 814
• 22 Hamada S, Slade HD. Biology, immunology, and cariogenicity of Streptococcus mutans . Microbiol Rev 1980; 44: 331-384
• 23 Smullen J, Koutsou G, Foster H, Zumbé A, Storey D. The antibacterial activity of plant extracts containing polyphenols against Streptococcus mutans . Caries Res 2007; 41: 342-349
• 24 Bubonja-Sonje M, Giacometti J, Abram M. Antioxidant and antilisterial activity of olive oil, cocoa and rosemary extract polyphenols. Food Chem 2011; 127: 1821-1827
• 25 Friedman M. Overview of antibacterial, antitoxin, antiviral, and antifungal activities of tea flavonoids and teas. Mol Nutr Food Res 2007; 51: 116-134
• 26 Ito K, Nakamura Y, Tokunaga T, Iijima D, Fukushima K. Anti-cariogenic properties of a water-soluble extract from cacao. Biosci Biotechnol Biochem 2003; 67: 2567-2573
• 27 Ferrazzano GF, Amato I, Ingenito A, De Natale A, Pollio A. Anti-cariogenic effects of polyphenols from plant stimulant beverages (cocoa, coffee, tea). Fitoterapia 2009; 80: 255-262
• 28 Banas J, Vickerman M. Glucan-binding proteins of the oral streptococci. Crit Rev Oral Biol Med 2003; 14: 89-99
• 29 Devulapalle K, Mooser G. Glucosyltransferase inactivation reduces dental caries. J Dent Res 2001; 80: 466-469
• 30 Li B, Li X, Lin H, Zhou Y. Curcumin as a promising antibacterial agent: effects on metabolism and biofilm formation in S. mutans . Biomed Res Int 2018; 2018: 4508709
• 31 Antonio AG, Moraes RS, Perrone D, Maia LC, Santos KRN, Iório NL, Farah A. Species, roasting degree and decaffeination influence the antibacterial activity of coffee against Streptococcus mutans . Food Chem 2010; 118: 782-788
• 32 Chen X, Mukwaya E, Wong MS, Zhang Y. A systematic review on biological activities of prenylated flavonoids. Pharm Biol 2014; 52: 655-660
• 33 Farhadi F, Khameneh B, Iranshahi M, Iranshahy M. Antibacterial activity of flavonoids and their structure-activity relationship: An update review. Phytother Res 2019; 33: 13-40
• 34 Lessa OA, dos Santos Reis N, Leite SGF, Gutarra MLE, Souza AO, Gualberto SA, de Oliveira JR, Aguiar-Oliveira E, Franco M. Effect of the solid state fermentation of cocoa shell on the secondary metabolites, antioxidant activity, and fatty acids. Food Sci Biotechnol 2018; 27: 107-113
• 35 Huang CB, Alimova Y, Myers TM, Ebersole JL. Short-and medium-chain fatty acids exhibit antimicrobial activity for oral microorganisms. Arch Oral Biol 2011; 56: 650-654
• 36 Desbois AP, Smith VJ. Antibacterial free fatty acids: activities, mechanisms of action and biotechnological potential. Appl Microbiol Biotechnol 2010; 85: 1629-1642
• 37 Barbosa-Pereira L, Rojo-Poveda O, Ferrocino I, Giordano M, Zeppa G. Assessment of volatile fingerprint by HS-SPME/GC-qMS and E-nose for the classification of cocoa bean shells using chemometrics. Food Res Int 2019; 123: 684-696
• 38 Barbosa-Pereira L, Rojo-Poveda O, Ferrocino I, Giordano M, Zeppa G. Analytical dataset on volatile compounds of cocoa bean shells from different cultivars and geographical origins. Data Brief 2019; 25: 104268
• 39 CLSI (Clinical and Laboratory Standards Institute). M07-A10: Methods for Dilution antimicrobial Susceptibility Tests for Bacteria that grow aerobically, Approved Standard – Tenth edition. Wayne, PA, USA: Clinical and Laboratory Standards Institute; 2015
• 40 Rojo-Poveda O, Barbosa-Pereira L, El Khattabi C, Youl ENH, Bertolino M, Delporte C, Pochet S, Stévigny C. Polyphenolic and methylxanthine bioaccessibility of cocoa bean shell functional biscuits: metabolomics approach and intestinal permeability through caco-2 cell models. Antioxidants (Basel) 2020; 9: 1164
• 41 Giacomoni F, Le Corguillé G, Monsoor M, Landi M, Pericard P, Pétéra M, Duperier C, Tremblay-Franco M, Martin JF, Jacob D. Workflow4Metabolomics: a collaborative research infrastructure for computational metabolomics. Bioinformatics 2015; 31: 1493-1495
• 42 Souard F, Delporte C, Stoffelen P, Thévenot EA, Noret N, Dauvergne B, Kauffmann JM, Van Antwerpen P, Stevigny C. Metabolomics fingerprint of coffee species determined by untargeted-profiling study using LC-HRMS. Food Chem 2018; 245: 603-612
• 43 Rinaudo P, Boudah S, Junot C, Thévenot EA. Biosigner: a new method for the discovery of significant molecular signatures from omics data. Front Mol Biosci 2016; 3: 26