Effect of Spoilage by Lipolytic Fungi Strains on Free Fatty Acids (FFA) Formation in Fermented and Dry Cocoa Beans

Tapé Joëlle Stéphanie, Houphouët Kouakou Richard, Koné Koumba Maï, M. Pauline, Guibert Benjamin, Métayer Isabelle, M. Didier, Guéhi Tagro Simplice
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Abstract

Aim: High free fatty acids (FFA) content of cocoa beans is high lead to alter quality of butter fraction and chocolate manufactured. During primary post-harvest processing, cocoa beans harbor lipase producing mold speciess could trigger the accumulation of FFA. This study investigated the improvement of the quality of raw cocoa material sourced from Côte d’Ivoire regarding its high FFA concentration. Methods: Fermented and dry cocoa were sampled on-farm level in some main cocoa producing regions of Côte d'Ivoire. Contaminating mold strains were isolated using Sabouraud chloramphenicol agar incubated at 25°C for 7 days. Morphological identification of mold isolates was preformed from younger fungal culture. Biochemical study about lipolytic properties of mold isolates was carried out using Cazpeck-Dox Agar medium enriched with olive oil as carbon source and rhodamine B as indicator of lipase activity. Extraction, PCR amplification using a specific transcribed spacer primer (ITS4/ITS5) and sequencing of DNA were performed for fungal species molecular identification. Results: Approximately 154 mold isolates belonging to 8 genera were found in tested cocoa beans. Only Rhizopus oryzae, Absidia corymbifera, Mucor heimalis, Aspergillus niger, A. tubingensis, A. aculeatus, A. flavus, A.  tamari, A. fumigatus, Nigrospora sphaerica, Curvularia geniculate and Penicillium chrysogenum species produced greater lipolytic properties. A. corymbifera, A. niger, A. fumigatus and R. orysae were the most frequent and predominant species. Cocoa beans inoculated with R. oryzae, A. corymbifera or A. niger recorded higher FFA content above 1.75%. Conclusion: Reducing of the mold species growth during cocoa primary post-harvest processing could sustainably produce raw cocoa material with less FFA.
溶脂真菌对发酵和干燥可可豆游离脂肪酸形成的影响
目的:可可豆中游离脂肪酸(FFA)含量高,是影响黄油和巧克力质量的主要因素。在采收后的初级加工过程中,可可豆港口脂肪酶产生霉菌可以触发游离脂肪酸的积累。本研究调查了产自Côte科特迪瓦的可可原料在其高游离脂肪酸浓度方面的质量改进。方法:在Côte科特迪瓦的一些主要可可产区进行农场水平的发酵可可和干可可取样。采用沙伯劳德氯霉素琼脂在25℃下孵育7 d分离污染霉菌菌株。从较年轻的真菌培养物中进行霉菌分离物的形态鉴定。以富含橄榄油的Cazpeck-Dox琼脂培养基为碳源,罗丹明B为脂肪酶活性指标,对霉菌分离物的脂解特性进行了生化研究。采用特异性转录间隔引物(ITS4/ITS5)进行提取、PCR扩增和DNA测序,对真菌种类进行分子鉴定。结果:在可可豆中分离出8属154株霉菌。只有米根霉、伞孢枯霉、黑毛霉、黑曲霉、塔binga .、acleatus、A. flavus、A. tamari、A. fumigatus、黑孢霉、弯曲曲霉和黄化青霉具有更强的溶脂性。恙螨、黑螨、烟螨和米螨是最常见和优势种。接种米曲霉、corymbifera和黑曲霉的可可豆中FFA含量均在1.75%以上。结论:在可可初级采收后加工过程中减少霉菌种类的生长,可以持续生产低游离脂肪酸的可可原料。
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