scholarly journals MICROBIAL SUCCESS OF ETHANOL, LACTIC ACID AND ACETIC ACID DURING FERMENTATION OF COCOA SEEDS

2019 ◽  
Author(s):  
mulono apriyanto bin sugeng rijanto

The objectives of the study are: 1) to determine the composition of the original cocoa bean pulp as a substrate for fermentation; 2) evaluating the effect of variations in random cocoa bean fermentation techniques on microbial populations. The stages of research carried out are as follows (1) testing the composition and moisture content of asalan cocoa beans as a fermentation substrate. (2) Fermented cocoa beans with 3 variations of fermentation techniques namely first treatment without addition of inoculum (control), second using S. cerevisiae (FNCC 3056) inoculum, L. lactis (FNC 0086) and A. aceti (FNCC 0016), respectively. - about 108 cfu / g is given simultaneously at the beginning of fermentation (IA). (3) gradual administration of inoculum yeast at the beginning of fermentation, lactic acid bacteria at 24 hours and acetic acid bacteria at 48 hours with a microbial population equal to the second treatment (IB). Fermentation is carried out for 120 hours. The temperature is set during fermentation, respectively 35 oC (first 24 hours), 45 oC (second 24 hours), 55 oC (third 24 hours) and 35 oC (last 48 hours). The results showed that during the fermentation of random cocoa beans showed that all treatments increased ethanol consumption in line with the increasing population of S. cerevisiae at the beginning of fermentation. Furthermore, L. lactis increases followed by lactic acid, at the end of A. aceti fermentation increases with acetic acid. From the results of this study it can be concluded that the rehydration of asalan cocoa beans can improve the composition of the pulp as a fermentation substrate. Microbial population shows that microbial succession has been demonstrated by the gradual addition of the inoculum.

2019 ◽  
Author(s):  
mulono apriyanto bin sugeng rijanto

The research objectives were: 1) to know the composition of cocoa bean pulp as substrate for fermentation; 2) evaluate the effect of variationof cocoa seed fermentation technique on microbial population. Stages of research conducted are as follows (1) testing the composition and water content of cocoa bean pulp as a fermentation substrate. (2) Fermented cocoa beans with 3 variations of fermentation technique ie first treatment withoutaddition of inoculum (control), both using inoculum S. cerevisiae (FNCC 3056), L. lactis (FNC 0086) and A. aceti (FNCC 0016), respectively - about 108cfu/g is given simultaneously at the beginning of fermentation (IA). (3) gradual inoculum administration of yeast at the begi nning of fermentation, lacticacid bacteria at 24 hours and acetic acid bacteria at 48 h with microbial population equal to second treatment (IB). Fermentation is carried out for 120hours. Temperatures are adjusted during fermentation, respectively 35 oC (first 24 hours), 45 oC (24 second hours), 55 oC (24 hours three) and 35 oC(last 48 hours). The third stage of fermented cocoa beans from the three treatments was roasted and analyzed for their volati le compounds. The resultsshowed that during the fermentation of cocoa beans showed that all treatments increased the ethanol kosentarsi in line with the increasing population ofS. cerevisiae at the beginning of fermentation. Next L. lactis increased followed by lactic acid, at the end of A. aceti fermentation increased followed byacetic acid. From the results of this study it can be concluded that the rehydration of cocoa bean pulp can improve the composition of pulp asfermentation substrate. The microbial population indicated that there was a microbial succession shown in the gradual addition of inoculum treatment.


2007 ◽  
Vol 73 (6) ◽  
pp. 1809-1824 ◽  
Author(s):  
Nicholas Camu ◽  
Tom De Winter ◽  
Kristof Verbrugghe ◽  
Ilse Cleenwerck ◽  
Peter Vandamme ◽  
...  

ABSTRACT The Ghanaian cocoa bean heap fermentation process was studied through a multiphasic approach, encompassing both microbiological and metabolite target analyses. A culture-dependent (plating and incubation, followed by repetitive-sequence-based PCR analyses of picked-up colonies) and culture-independent (denaturing gradient gel electrophoresis [DGGE] of 16S rRNA gene amplicons, PCR-DGGE) approach revealed a limited biodiversity and targeted population dynamics of both lactic acid bacteria (LAB) and acetic acid bacteria (AAB) during fermentation. Four main clusters were identified among the LAB isolated: Lactobacillus plantarum, Lactobacillus fermentum, Leuconostoc pseudomesenteroides, and Enterococcus casseliflavus. Other taxa encompassed, for instance, Weissella. Only four clusters were found among the AAB identified: Acetobacter pasteurianus, Acetobacter syzygii-like bacteria, and two small clusters of Acetobacter tropicalis-like bacteria. Particular strains of L. plantarum, L. fermentum, and A. pasteurianus, originating from the environment, were well adapted to the environmental conditions prevailing during Ghanaian cocoa bean heap fermentation and apparently played a significant role in the cocoa bean fermentation process. Yeasts produced ethanol from sugars, and LAB produced lactic acid, acetic acid, ethanol, and mannitol from sugars and/or citrate. Whereas L. plantarum strains were abundant in the beginning of the fermentation, L. fermentum strains converted fructose into mannitol upon prolonged fermentation. A. pasteurianus grew on ethanol, mannitol, and lactate and converted ethanol into acetic acid. A newly proposed Weissella sp., referred to as “Weissella ghanaensis,” was detected through PCR-DGGE analysis in some of the fermentations and was only occasionally picked up through culture-based isolation. Two new species of Acetobacter were found as well, namely, the species tentatively named“ Acetobacter senegalensis” (A. tropicalis-like) and “Acetobacter ghanaensis” (A. syzygii-like).


2021 ◽  
Author(s):  
Romel E. Guzmán-Alvarez ◽  
José G. Márquez-Ramos

Cocoa bean fermentation is a spontaneous process driven by an ordered microbial succession of a wide range of yeasts, lactic acid and acetic acid bacteria, some aerobic sporeforming bacteria and various species of filamentous fungi. The process of cocoa fermentation is a very important step for developing chocolate flavor precursors which are attributable to the metabolism of succession microbial. The microbial ecology of cocoa has been studied in much of the world. In Venezuela, studies have been carried out with Criollo, Forastero, and Trinitario cocoa, fermented under various conditions, the results obtained coinciding with the reported scientific information. Fermentation must be associated with the type of cocoa available, carried out knowing the final processing and derivative (paste, butter, powder). The results shown in this chapter correspond to investigations carried out with cocoa from three locations in Venezuela. The quantification, identification, isolation, functionality of the most representative microbiota involved in the fermentation of these grains was sought. This to give possible answers to the fermentation times and improvement of the commercial quality. Likewise, generate greater interest on the part of the producers in carrying out the fermentation.


Fermentation ◽  
2021 ◽  
Vol 7 (1) ◽  
pp. 42
Author(s):  
Lucie Farrera ◽  
Alexandre Colas de la Noue ◽  
Caroline Strub ◽  
Benjamin Guibert ◽  
Christelle Kouame ◽  
...  

Acetic acid bacteria are involved in many food and beverage fermentation processes. They play an important role in cocoa bean fermentation through their acetic acid production. They initiate the development of some of the flavor precursors that are necessary for the organoleptic quality of cocoa, and for the beans’ color. The development of starter cultures with local strains would enable the preservation of the microbial biodiversity of each country in cocoa-producing areas, and would also control the fermentation. This approach could avoid the standardization of cocoa bean fermentation in the producing countries. One hundred and thirty acetic acid bacteria were isolated from three different cocoa-producing countries, and were identified based on their 16S rRNA gene sequence. The predominate strains were grown in a cocoa pulp simulation medium (CPSM-AAB) in order to compare their physiological traits regarding their specific growth rate, ethanol and lactic acid consumption, acetic acid production, and relative preferences of carbon sources. Finally, the intraspecific diversity of the strains was then assessed through the analysis of their genomic polymorphism by (GTG)5-PCR fingerprinting. Our results showed that Acetobacter pasteurianus was the most recovered species in all of the origins, with 86 isolates out of 130 cultures. A great similarity was observed between the strains according to their physiological characterization and genomic polymorphisms. However, the multi-parametric clustering results in the different groups highlighted some differences in their basic metabolism, such as their efficiency in converting carbon substrates to acetate, and their relative affinity to lactic acid and ethanol. The A. pasteurianus strains showed different behaviors regarding their ability to oxidize ethanol and lactic acid into acetic acid, and in their relative preference for each substrate. The impact of these behaviors on the cocoa quality should be investigated, and should be considered as a criterion for the selection of acetic acid bacteria starters.


Fermentation ◽  
2021 ◽  
Vol 7 (3) ◽  
pp. 192
Author(s):  
Endang Sutriswati Rahayu ◽  
Rokhmat Triyadi ◽  
Rosyida N. B. Khusna ◽  
Titiek Farianti Djaafar ◽  
Tyas Utami ◽  
...  

Cocoa bean fermentation is an important process in the manufacturing of cocoa products. It involves microbes, such as lactic acid bacteria, yeast, and acetic acid bacteria. The presence of mold in cocoa bean fermentation is undesired, as it reduces the quality and may produce mycotoxins, which can cause poisoning and death. Aspergillus niger is a fungus that produces ochratoxin A, which is often found in dried agricultural products such as seeds and cereals. In this study, we applied indigenous Candida famata HY-37, Lactobacillus plantarum HL-15, and Acetobacter spp. HA-37 as starter cultures for cocoa bean fermentation. We found that the use of L. plantarum HL-15 individually or in combination Candida famata HY-37, Lactobacillus plantarum HL-15, and Acetobacter spp. HA-37 as a starter for cocoa bean fermentation can inhibit the growth of A. niger YAC-9 and the synthesis of ochratoxin A during fermentation and drying. With biological methods that use indigenous Lactobacillus plantarum HL-15 individually or in combination with Candida famata HY-37 and Acetobacter spp. HA-37, we successfully inhibited contamination by ochratoxin-A-producing fungi. Thus, the three indigenous microbes should be used in cocoa bean fermentation to inhibit the growth of fungi that produce mycotoxins and thus improve the quality.


2010 ◽  
Vol 76 (23) ◽  
pp. 7708-7716 ◽  
Author(s):  
Timothy Lefeber ◽  
Maarten Janssens ◽  
Nicholas Camu ◽  
Luc De Vuyst

ABSTRACT The composition of cocoa pulp simulation media (PSM) was optimized with species-specific strains of lactic acid bacteria (PSM-LAB) and acetic acid bacteria (PSM-AAB). Also, laboratory fermentations were carried out in PSM to investigate growth and metabolite production of strains of Lactobacillus plantarum and Lactobacillus fermentum and of Acetobacter pasteurianus isolated from Ghanaian cocoa bean heap fermentations, in view of the development of a defined starter culture. In a first step, a selection of strains was made out of a pool of strains of these LAB and AAB species, obtained from previous studies, based on their fermentation kinetics in PSM. Also, various concentrations of citric acid in the presence of glucose and/or fructose (PSM-LAB) and of lactic acid in the presence of ethanol (PSM-AAB) were tested. These data could explain the competitiveness of particular cocoa-specific strains, namely, L. plantarum 80 (homolactic and acid tolerant), L. fermentum 222 (heterolactic, citric acid fermenting, mannitol producing, and less acid tolerant), and A. pasteurianus 386B (ethanol and lactic acid oxidizing, acetic acid overoxidizing, acid tolerant, and moderately heat tolerant), during the natural cocoa bean fermentation process. For instance, it turned out that the capacity to use citric acid, which was exhibited by L. fermentum 222, is of the utmost importance. Also, the formation of mannitol was dependent not only on the LAB strain but also on environmental conditions. A mixture of L. plantarum 80, L. fermentum 222, and A. pasteurianus 386B can now be considered a mixed-strain starter culture for better controlled and more reliable cocoa bean fermentation processes.


2012 ◽  
Vol 78 (15) ◽  
pp. 5395-5405 ◽  
Author(s):  
Gilberto Vinícius de Melo Pereira ◽  
Maria Gabriela da Cruz Pedrozo Miguel ◽  
Cíntia Lacerda Ramos ◽  
Rosane Freitas Schwan

ABSTRACTSpontaneous cocoa bean fermentations performed under bench- and pilot-scale conditions were studied using an integrated microbiological approach with culture-dependent and culture-independent techniques, as well as analyses of target metabolites from both cocoa pulp and cotyledons. Both fermentation ecosystems reached equilibrium through a two-phase process, starting with the simultaneous growth of the yeasts (withSaccharomyces cerevisiaeas the dominant species) and lactic acid bacteria (LAB) (Lactobacillus fermentumandLactobacillus plantarumwere the dominant species), which were gradually replaced by the acetic acid bacteria (AAB) (Acetobacter tropicaliswas the dominant species). In both processes, a sequence of substrate consumption (sucrose, glucose, fructose, and citric acid) and metabolite production kinetics (ethanol, lactic acid, and acetic acid) similar to that of previous, larger-scale fermentation experiments was observed. The technological potential of yeast, LAB, and AAB isolates was evaluated using a polyphasic study that included the measurement of stress-tolerant growth and fermentation kinetic parameters in cocoa pulp media. Overall, strainsL. fermentumUFLA CHBE8.12 (citric acid fermenting, lactic acid producing, and tolerant to heat, acid, lactic acid, and ethanol),S. cerevisiaeUFLA CHYC7.04 (ethanol producing and tolerant to acid, heat, and ethanol), andAcetobacter tropicalisUFLA CHBE16.01 (ethanol and lactic acid oxidizing, acetic acid producing, and tolerant to acid, heat, acetic acid, and ethanol) were selected to form a cocktail starter culture that should lead to better-controlled and more-reliable cocoa bean fermentation processes.


Author(s):  
Maria Denis Lozano Tovar ◽  
Geraldine Tibasosa ◽  
Carlos Mario González ◽  
Karen Ballestas Alvarez ◽  
Martha Del Pilar Lopez Hernandez ◽  
...  

Microbial activity involved in the cocoa beans fermentation process is essential to maintain and improve the organoleptic and nutritional qualities of chocolate; therefore, the aim of this investigation was to search and select microbial isolates with the potential to improve the quality of cocoa beans. Fermentation experimentswere conducted on farms located in Maceo (Antioquia), San Vicente de Chucurí (Santander), and Rivera and Algeciras (Huila), Colombia. Yeast, lactic acid bacteria (LAB), acetic acid bacteria (AAB), and mesophilic aerobic microorganisms were obtained from different fermentation batches. The growth of these microorganismswas tested in six treatments as follows: 50% cocoa pulp agar (CPA), high concentrations of glucose (10%), ethanol (5%), and acetic acid (7%), an acidic pH of 3.0, and a high temperature of 50oC for 24 h. The isolates with the highest growth were identified by 18S and 16S rRNA gene analysis, revealing a high diversity ofspecies associated with cocoa fermentation, including eight species of yeasts (Debaryomyces hansenii, Meyerozyma guillermondii, Wickerhanomyces anomalus, Pichia guillermondii, Pichia kudriavzevii, Trichosporon asahii, Candida parapsilosis, and Pichia manshurica), six species of LAB (Pediococcus acidilactici, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus farraginis, Lactobacillus rhamnosus, and Leuconostoc mesenteroides), four species of AAB (Gluconobacter japonicus, Acetobacter tropicalis, Acetobacter pasteurianus, and Acetobacter malorum/tropicalis), and three species of Bacillus spp. (Bacillusaryabhattai /megaterium, Bacillus subtilis, and Bacillus coagulans). In general, microbial populations increased in cocoa batches after 12 h of fermentation and decreased after 84-96 h. All the yeast isolates grew in 10% glucose and CPA, 85.7% in 5% ethanol, and 95% at a pH of 3.0. All the yeast isolates were affectedby 7% acetic acid and incubation at 50oC for 24 h. Eighty-five percent of the LAB grew in 10% glucose, 100% in 5% ethanol, 42.8% in CPA, 64% at a pH of 3.0, and 35.7% grew after being exposed to 50oC for 24 h; all were affected by 7% acetic acid. As for the AAB, 100% grew in 10% glucose, 71% in 7% ethanol, 100% grew in CPA, in 7% acetic acid, and at a pH of 3.0, while 100% were affected by incubation at 50oC. Three yeast isolates, W. anomalus, D. hansenii and M. guillermondii, three LAB isolates, P. acidilactici, L. brevis, and L. plantarum, and three AAB isolates, A. tropicalis, A. pasteurianus and G. japonicus, were selected as promising strains to be used in a microbial starter culture for cocoa bean fermentation to improve the organoleptic quality of cocoa.


Author(s):  
O.G.G. Almeida ◽  
E.C.P De Martinis

Metagenomic studies about cocoa fermentation have mainly reported on the analysis of short reads for determination of Operational Taxonomic Units. However, it is also important to determine MAGs, which are genomes deriving from the assembly of metagenomics. For this research, all the cocoa metagenomes from public databases were downloaded, resulting in five datasets: one from Ghana and four from Brazil. Besides, in silico approaches were used to describe putative phenotypes and metabolic potential of MAGs. A total of 17 high-quality MAGs were recovered from these microbiomes, as follows: (i) fungi - Yamadazyma tenuis (n=1); (ii) lactic acid bacteria - Limosilactobacillus fermentum (n=5), Liquorilactobacillus cacaonum (n=1) , Liquorilactobacillus nagelli (n=1), Leuconostoc pseudomesenteroides (n=1) and Lactiplantibacillus plantarum subsp. plantarum (n=1); (iii) acetic acid bacteria - Acetobacter senegalensis (n=2) and Kozakia baliensis (n=1) and (iv) Bacillus subtilis (n=1) Brevundimonas sp. (n=2) and Pseudomonas sp. (n=1). Medium-quality MAGs were also recovered from cocoa microbiomes, including some detected for the first time in this environment ( Liquorilactobacillus vini , Komagataeibacter saccharivorans and Komagataeibacter maltaceti ) and other previously described ( Fructobacillus pseudoficulneus and Acetobacter pasteurianus ). Taken all together, the MAGs were useful to provide an additional description of the microbiome of cocoa fermentation, revealing previously overlooked microorganisms, with prediction of key phenotypes and biochemical pathways. Importance The production of chocolate starts with the harvesting of cocoa fruits and the spontaneous fermentation of the seeds, in a microbial succession that depends on yeasts, lactic acid bacteria and acetic acid bacteria in order to eliminate bitter and astringent compounds present in the raw material, which will be further roasted and grinded to originate the cocoa powder that will enter the food processing industry. The microbiota of cocoa fermentation is not completely know, and yet it advanced from culture-based studies to the advent of Next Generation DNA sequencing, with the generation of a myriad of data, that need bioinformatic approaches to be properly analysed. Although the majority metagenomic of studies have been based on short reads (OTUs), it is also important to analyse entire genomes to determine more precisely possible ecological roles of different species. Metagenome-assembled genomes (MAGs) are very useful for this purpose, and in this paper, MAGs from cocoa fermentation microbiomes were described, as well the possible implications of their phenotypic and metabolic potentials are discussed.


Sign in / Sign up

Export Citation Format

Share Document