Effect of Fermentation on Bacteria Isolates and Phytochemical Properties of Cocoa (Theobroma cacao) Beans

Main Article Content

F. O. Ibitoye
A. R. Osaloni
V. T. Adebote


This investigation was carried out to assess the bacteria associated with cocoa beans at different stages of fermentation and determine the changes in the phytochemical constituents of the fermenting beans. The pour plate technique was used for bacterial isolation while phytochemicals were assessed based on standard qualitative chemical reactions. The total bacterial count on the cocoa beans reduced during fermentation from the initial 86.2±0.02 x 105 CFU/g (day 0) to 1.00±0.00 x 105 CFU/g on day 5. However, there was an increase in the lactic acid bacteria count from 48.7±0.03 x 105 CFU/g (day 0) to 111.7±0.03 x 105 CFU/g on day 3, then reduced to 51.4±0.01 x 105 CFU/g on day 5. Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus acidophilus were isolated from the cocoa beans at different stages of the fermentation. There was a gradual increase in the temperature of the fermenting cocoa mass from the initial 25.6°C recorded at the beginning of the fermentation to 42.8°C recorded on day 5. Also, the pH of the fermenting cocoa beans reduced significantly from 6.1 at the commencement of the fermentation to 3.2 on day 5. In the total titratable acidity assay, the acidity of the cocoa beans increased from 3.12% at the beginning of the fermentation to 7.12% on day 5. Further, in the phytochemical screening, only alkaloids, phenols, steroids and flavonoids were detected in the beans throughout fermentation period whereas tannin and saponin were not found in the beans at any stage of fermentation. The preset phytochemicals got reduced in intensity as fermentation advanced. From these results, it can therefore be concluded that fermentation helps to improve the taste quality and phytochemical properties of Nigerian cocoa beans.

Cocoa, microorganisms, fermentation, metabolites

Article Details

How to Cite
Ibitoye, F. O., Osaloni, A. R., & Adebote, V. T. (2020). Effect of Fermentation on Bacteria Isolates and Phytochemical Properties of Cocoa (Theobroma cacao) Beans. South Asian Journal of Research in Microbiology, 7(4), 1-8. https://doi.org/10.9734/sajrm/2020/v7i430176
Original Research Article


Afoakwa E, Paterson A, Fowler M. Factors influencing rheological and textural qualities in chocolate e a review. Trends in Food Science and Technology. 2014; 18(6):290-298.

Afoakwa EO, Paterson A, Fowler M, Ryan A. Flavor formation and character in cocoa and chocolate: A critical review. Critical Reviews in Food Science and Nutrition. 2008;48:840–857.

Serra Bonvehí J, Ventura Coll F. Factors affecting the formation of alkylpyrazines during roasting treatment in natural and alkalinized cocoa powder. Journal of Agricultural and Food Chemistry. 2002; 50(13):3743-3750.

Lima LJR, Almeida MH, Nout MJR, Zwietering MH. Theobroma cacao L., the food of the gods: quality determinants of commercial cocoa beans, with particular reference to the impact of the fermentation. Critical Reviews in Food Science and Nutrition. 2011;51(8):731-761.

Rohsius C, Matissek R, Lieberei R. Free amino acid amounts in raw cocoas from different origins. European Food Research and Technology. 2006;222:432-438.

Arlorio M, Locatelli M, Travaglia F, Coïsson JD, Del Grosso E, Minassi A. Roasting impact on the contents of clovamide (N-caffeoyl-L-DOPA) and the antioxidant activity of cocoa beans (Theobroma cacao L.). Food Chemistry. 2008;106(3):967-975.

Sukha DA, Butler DR, Umaharan PE, Boult E. The use of an optimized organoleptic assessment protocol to describe and quantify different flavour attributes of cocoa liquors made from Ghana and Trinitario beans. European Journal of Food and Technology. 2008; 226:405–413.

Ardhana M, Fleet G. The microbial ecology of cocoa bean fermentations in Indonesia. International Journal of Food Microbiology. 2003;86:87–99.

de Brito ES, García NHP, Gallão MI, Cortelazzo AL, Fevereiro PS, Braga MR. Structural and chemical changes in cocoa (Theobroma cacao L) during fermentation, drying and roasting. Journal of the Science of Food and Agriculture. 2001;81(2):281-288.

Camu N, De Winter T, Addo SK, Takarama JS, Bernaert H, De Vuyst L. Fermentation of cocoa beans: Influence of microbial activities and polyphenol concentrations on the flavour of chocolate. Journal of the Science of Food and Agriculture. 2008;88:2288–2297.

Edeoga HO, Eriata DO, Alkaloid tannin. Saponin contents of some Nigeria medicinal plants. Journal of Medicinal Aromatic Plant Science. 2001;23:344-349.

Ogundipe OO, Moody JO, Houghton PJ, Odelola HA. Bioactive chemical constituents from Alchornea laxiflora (Benth.) Pax and Hoffman. J. Ethno-pharmacol. 2001;74:275-280.

Misnawi S. Physic-chemical changes during cocaoa fermentation and key enzymes involved. Review penelitian kopi dan kakao 2008;24(1):54-71.

Papalexandratou Z, Vranckena G, De Bruyneb K, Vandammeb P, De Vuys L. Spontaneous organic cocoa bean box fermentations in Brazil are characterized by a restricted species diversity of lactic acid bacteria and acetic acid bacteria. Food Microbiology. 2011;28:1326–1338.

Association of Official Analytical Chemist. Official methods of analysis, 18th Ed. Washington DC; 2005.

Sofowora A. Phytochemical screening of medicinal plants and traditional medicine in Africa. 2nd edition Spectrum Books Limited, Nigeria. 1993;150–156.

Schwan RF, Wheals AE. The microbiology of cocoa fermentation and its role in chocolate quality. Critical Reviews in Food Science and Nutrition. 2014;44:205–221.

Carr J, Davies P, Dougan J. Cocoa fermentation in Ghana and Malaysia. In 7th International Cocoa Research Conference. 2009:573-576.

LagunesGálvez S, Loiseau G, Paredes JL, Barel M, Guiraud JP. Study on the microflora and biochemistry of cocoa fermentation in the Dominican Republic. International Journal of Food Microbiology. 2007;114(1):124-130.

Nazaruddin R, Seng L, Hassan O, Said M. Effect of pulp preconditioning on the content of polyphenols in cocoa beans (Theobroma cacao) during fermentation. Industrial Crops and Products. 2006; 24(1):87-94.

Afoakwa EO, Quao J, Takrama J, Budu AS, Saalia FK. Effect of pulp preconditioning on acidification, proteolysis of sugars and free fatty acids concentration during fermentation of cocoa beans. Intr J. of Food science and nutrition. 2011;62(7):755-764.

Wollgast J, Anklam E. Review on polyphenols in Theobroma cacao: changes in composition during the manufacture of chocolate and methodology for identification and quantification. Food Research International. 2000;33:423-447.