Cocoa Residues as Support Material for Acetobacter pasteurianus Starter Culture Freeze-Drying

Lamine SAMAGACI^1, , Hadja OUATTARA¹, Victoria KADET¹, Syntyche BOGUI², Théodore DJENI² and Sébastien NIAMKE¹

¹Biotechnology Laboratory, UFR Biosciences, Félix HOUPHOUËT-BOIGNY University, Abidjan, Côte d'Ivoire

²Food Biotechnology and Microbiology Laboratory, UFR Sciences and Food Technology, Nangui Abrogua University, Abidjan, Côte d'Ivoire

Cite this paper:
Lamine SAMAGACI, Hadja OUATTARA, Victoria KADET, Syntyche BOGUI, Théodore DJENI and Sébastien NIAMKE. Cocoa Residues as Support Material for Acetobacter pasteurianus Starter Culture Freeze-Drying. Journal of Applied & Environmental Microbiology. 2025; 13(1):7-11. doi: 10.12691/jaem-13-1-2

Abstract

Microbial starters are of great economic importance to the food industry and many other sectors. These organisms ensure that food is processed to produce reproducible products of consistent quality. Maintaining their stability and viability during processing and storage is therefore a priority. Of all the methods proposed for their preservation, freeze-drying remains the most suitable for bacteria. In this study, cocoa pod flour and/or pulp was used as a cryoprotectant, in comparison with mannitol, the cryoprotectant that best protects acetic bacteria. Acetobacter pasteurianus strains were grown in YEPG broth and centrifuged. The pellets were then collected and mixed with 20% mannitol, cocoa pulp juice, cocoa pericarp flour, and a combination of cocoa pulp juice and cocoa pericarp flour. The samples were freeze-dried, and the survival rate after freeze-drying and during storage as well as acid production were assessed. The results show that the combination of cocoa pericarp and cocoa pulp juice preserved the strains better during lyophilization and storage (79.24 ± 0.32%) than the control (76.05 ± 0.37%) and the other trials. In addition, the acid production of the strain was better preserved by the combination of cocoa pericarp flour and cocoa pulp juice than in any of the other trials. Cocoa pericarp flour combined with cocoa pulp juice could be used as an alternative cryoprotectant for freeze-drying of Acetobacter pasteurianus. This could enable the valorisation of cocoa residues, such as pod and pulp.

Keywords:
Acetobacter pasteurianus cocoa pulp cocoa pericarp cryoprotectants

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References:

[1]	Raspor P., Goranovič D., "Biotechnological applications of acetic acid bacteria", Critical reviews in biotechnology, 28 (2), 101-24, 2008.
[2]	Lynch K.M., Zannini E., Wilkinson S., Daenen L., Arendt E.K., "Physiology of acetic acid bacteria and their role in vinegar and fermented beverages", Comprehensive reviews in food science and food safety, 18 (3), 587-625, 2019.
[3]	Mizzi J., Gaggìa F., Bozzi Cionci N., Di Gioia D., Attard E., "Selection of Acetic Acid Bacterial Strains and Vinegar Production From Local Maltese Food Sources", Frontiers in Microbiology, 13, 897825, 2022.
[4]	Román-Camacho J.J., García-García I., Santos-Dueñas I.M., García-Martínez T., Mauricio J.C., "Latest trends in industrial vinegar production and the role of acetic acid bacteria: classification, metabolism, and applications—a comprehensive review", Foods, 12 (19), 3705, 2023.
[5]	Sengun I.Y., Kilic G., Charoenyingcharoen P., Yukphan P., Yamada Y., "Investigation of the microbiota associated with traditionally produced fruit vinegars with focus on acetic acid bacteria and lactic acid bacteria", Food Bioscience, 47, 101636, 2022.
[6]	Bartowsky E.J., Henschke P.A., "Acetic acid bacteria spoilage of bottled red wine—A review", International Journal of Food Microbiology, 125 (1), 60-70, 2008.
[7]	De Vero L., Giudici P., "Genus-specific profile of acetic acid bacteria by 16S rDNA PCR-DGGE", International Journal of Food Microbiology, 125 (1), 96-101, 2008.
[8]	Sengun I.Y., Karabiyikli S., "Importance of acetic acid bacteria in food industry", Food control, 22 (5), 647-56, 2011.
[9]	Yassunaka Hata N.N., Surek M., Sartori D., Vassoler Serrato R., Aparecida Spinosa W., "Role of acetic acid bacteria in food and beverages", Food technology and biotechnology, 61 (1), 85-103, 2023.
[10]	Coulibaly P., Goualié B., Samagaci L., Ouattara H., Niamké S., "Screening of thermotolerant acetic acid bacteria involved in cocoa fermentation in six major cocoa producing regions in côte d’ivoire", Biotechnology Journal International, 21 (2), 1-15, 2018.
[11]	Soumahoro S., Ouattara H.G., Droux M., Nasser W., Niamke S.L., Reverchon S., "Acetic acid bacteria (AAB) involved in cocoa fermentation from Ivory Coast: species diversity and performance in acetic acid production", Journal of food science and technology, 57, 1904-16, 2020.
[12]	Deppenmeier U., Hoffmeister M., Prust C., "Biochemistry and biotechnological applications of Gluconobacter strains", Applied Microbiology and Biotechnology, 60, 233-42, 2002.
[13]	Bossart L., "Contribution à l’optimisation du séchage en lit fluidisé. Thèse, Université Libre de Bruxelles, Belgique.", 2006.
[14]	Bossart L., Halloin V., "Séchage des levures en lit fluidisé. Chemical Engineering and Processing, 45, 1019–1028. ", 2006.
[15]	Santivarangkna C., Kulozik U., Foerst P., "Alternative drying processes for the industrial preservation of lactic acid starter cultures", Biotechnology progress, 23 (2), 302-15, 2007.
[16]	Ndoye B., Weekers F., Diawara B., Guiro A.T., Thonart P., "Survival and preservation after freeze-drying process of thermoresistant acetic acid bacteria isolated from tropical products of Subsaharan Africa", Journal of food engineering, 79 (4), 1374-82, 2007.
[17]	Santivarangkna C., Higl B., Foerst P., "Protection mechanisms of sugars during different stages of preparation process of dried lactic acid starter cultures", Food microbiology, 25 (3), 429-41, 2008.
[18]	Kadet V., Samagaci L., Ouattara H., Ahoussi J.-M., Ettien Y., Niamké S., "Soy Flour, a Support for Freeze Drying of Acetobacter Pasteurianus Starter for Cocoa Fermentation in Côte d’Ivoire", European Journal of Agriculture and Food Sciences, 6 (3), 27-32, 2024.
[19]	Dubois M., "Use of phenol reagent for the determination of total sugar", Anal Chem, 28, 350, 1956.
[20]	Bernfeld P., " Amylases α and β. In: Methods in Enzymology, ed. By Colowick S.P. and Kalpan N. O., Academic Press: New York, (1): 149-158", 1955.
[21]	Cui S., Hu M., Sun Y., Mao B., Zhang Q., Zhao J., et al., "Effect of Trehalose and Lactose Treatments on the Freeze-Drying Resistance of Lactic Acid Bacteria in High-Density Culture", Microorganisms, 11 (1), 48, 2022.
[22]	Mendoza G.M., Pasteris S.E., Otero M.C., Nader-Macias F.M.E., "Survival and beneficial properties of lactic acid bacteria from raniculture subjected to freeze-drying and storage", Journal of Applied Microbiology, 116, 157-66, 2013.
[23]	Martín M.J., Lara-Villoslada F., Ruiz M.A., Morales M.E., "Microencapsulation of bacteria: A review of different technologies and their impact on the probiotic effects", Innovative Food Science & Emerging Technologies, 27, 15-25, 2015.
[24]	Coulibaly H.W. Mise en place d'un starter lyophilisé pour la fermentation alcoolique de la biere de sorgho. Ph.D. Thesis, Université Nangui Abrogoua, Côte d'Ivoire, 2016.
[25]	Leite P.B., Maciel L.F., Opretzka L.C.F., Soares S.E., Bispo E.d.S., "Phenolic compounds, methylxanthines and antioxidant activity in cocoa mass and chocolates produced from" witch broom disease" resistant and non resistant cocoa cultivars", Ciência e agrotecnologia, 37, 244-50, 2013.
[26]	Celik O., O’Sullivan D., "Factors influencing the stability of freeze-dried stress-resilient and stress-sensitive strains of bifidobacteria", Journal of dairy science, 96 (6), 3506-16, 2013.
[27]	Chen B., Wang X., Li P., Feng X., Mao Z., Wei J., et al., "Exploring the protective effects of freeze-dried Lactobacillus rhamnosus under optimized cryoprotectants formulation", LWT-Food Science and Technology, 173, 114295, 2023.
[28]	Turner S., Senaratna T., Touchell D., Bunn E., Dixon K., Tan B., "Stereochemical arrangement of hydroxyl groups in sugar and polyalcohol molecules as an important factor in effective cryopreservation", Plant Science, 160 (3), 489-97, 2001.
[29]	Shafiei R., Delvigne F., Thonart P., "Flow-cytometric assessment of damages to Acetobacter senegalensis during freeze-drying process and storage", Acetic Acid Bacteria, 2 (1), e10-e, 2013.
[30]	Lei P., Chen H., Ma J., Fang Y., Qu L., Yang Q., et al., "Research progress on extraction technology and biomedical function of natural sugar substitutes", Frontiers in Nutrition, 9, 952147, 2022.
[31]	Valladares-Diestra K.K., de Souza Vandenberghe L.P., Soccol C.R., "A biorefinery approach for pectin extraction and second-generation bioethanol production from cocoa pod husk", Bioresource technology, 346, 126635, 2022.
[32]	de Souza Vandenberghe L.P., Valladares-Diestra K.K., Bittencourt G.A., de Mello A.F.M., Vásquez Z.S., de Oliveira P.Z., et al., "Added-value biomolecules’ production from cocoa pod husks: A review", Bioresource technology, 344, 126252, 2022.
[33]	Bickel Haase T., Schweiggert-Weisz U., Ortner E., Zorn H., Naumann S., "Aroma properties of cocoa fruit pulp from different origins", Molecules, 26 (24), 7618, 2021.
[34]	Rajan R., Matsumura K., "Development and application of cryoprotectants", Advances in Experimental Medicine and Biology, 1081, 339-54, 2018.
[35]	Jocelyne R.E., Béhiblo K., Ernest A.K., "Comparative study of nutritional value of wheat, maize, sorghum, millet, and fonio: some cereals commonly consumed in Côte d’Ivoire", European Scientific Journal, 16 (21), 118-31, 2020.