Effects of incubation and polyphenol oxidase enrichment on colour, fermentation index, procyanidins and astringency of unfermented and partly fermented cocoa beans

Incubation of unfermented and partly fermented cocoa beans in acetate buffer, pH 5.5, at 45 °C increased yellowness, total colour differences and fermentation index value of the cocoa bean powders and decreased cocoa procyanidins (monomers to pentamers), and their astringency. Fermentation index and (–)‐epicatechin content, equivalent to those of fully fermented beans, were reached by unfermented beans after 4–8‐h incubation, but not by partly fermented beans even after 16 h. During incubation of partly fermented cocoa beans enriched with polyphenol oxidase, yellowness and fermentation index value were increased, whilst (–)‐epicatechin was decreased. Tyrosinase had a less significant effect in yellow colour formation, but showed a significant reduction of (–)‐epicatechin and increase in fermentation index compared with crude cocoa polyphenol oxidase. However, both enzymes have similar effects on procyanidin degradation and astringent taste reduction. Incubation of cocoa beans for 16 h increased the cut test score of unfermented and partly fermented beans by 50 and 30%, respectively.     

Oxidation of polyphenols in unfermented and partly fermented cocoa beans by cocoa polyphenol oxidase and tyrosinase

Unfermented and partly fermented dried cocoa beans have an excessively astringent and bitter flavour owing to their high polyphenol content. Studies on the remaining polyphenol oxidase activity and the oxidation of polyphenols in these beans have been conducted in relation to two factors, ie incubation time (0, 1, 2, 4, 8 and 16 h) and pH of incubation (3.5, 4.5, 5.5 and 6.5). Owing to unsuccessful polyphenol oxidation during incubation of partly fermented beans, incubation–enrichment treatments were carried out by combining incubation time (0, 8, 16, 24 and 32 h) and addition of crude cocoa polyphenol oxidase and purified tyrosinase from mushroom at 88 and 8800 units g^?1 and pH 5.5. Results showed that the remaining polyphenol oxidase activities of unfermented and partly fermented dried beans were 1 and 0.08% with specific activities of 9 and 1% respectively. The polyphenols of unfermented beans were effectively oxidised by incubation at 45 °C and pH 3.5–6.5 without enzyme enrichment; while those of partly fermented beans required enzyme enrichment. Both crude cocoa polyphenol oxidase and tyrosinase could be used for enzyme enrichment, but tyrosinase seemed to be more effective. © 2002 Society of Chemical Industry     

Aroma precursors and methylpyrazines in underfermented cocoa beans induced by endogenous carboxypeptidase

Cocoa-specific aroma precursors and methylpyrazines in underfermented cocoa beans obtained from fermentation induced by indigenous carboxypeptidase have been investigated. Fermentation conditions and cocoa bean components were analyzed during 0 to 3 d of fermentation. Underfermented cocoa beans were characterized as having hydrophilic peptides and free hydrophobic amino acids much higher than unfermented ones. These 2 key components of cocoa aroma precursors may be produced from the breakdown of proteins and polypeptides by endogenous carboxypeptidase during the fermentation process. The enzyme was activated during fermentation. Polypeptides of 47, 31, and 19 kDa were observed in the samples throughout the 3-d fermentation period; however, only the first 2 polypeptides were remarkably reduced during fermentation. Since the 1st day of fermentation, underfermented cocoa beans contained methylpyrazines, a dominant group of cocoa-specific aroma. This might be due to microbial activities during fermentation, observed through a decrease of pH value and an increase of temperature of cocoa beans. The concentration of tetramethylpyrazines was significantly increased during the 3 d of fermentation. This may increase the cocoa-specific flavor to the beans.     

Comparison of enzyme activities involved in flavour precursor formation in unfermented beans of different cocoa genotypes

The activities of endoprotease, aminopeptidase, carboxypeptidase and invertase (cotyledon and pulp) were studied in unfermented beans of 10 genotype samples with different flavour characteristics (high and low cocoa flavour). Analysis of variance showed that significant differences in enzyme activities exist between certain genotypes. Aminopeptidase and endoprotease activities in beans of the PA7 genotype were higher than in all others. Principal component analysis (PCA) showed that the PA7 genotype (high cocoa flavour) was very different from the UIT1 genotype (low cocoa flavour). Although significant differences exist, no simple and general relationship is established between the flavour potential of a genotype and the level of key enzyme activities in unfermented beans. Carboxypeptidase is of key importance for peptide and free amino acid formation, but differences in enzyme activity could not be correlated to flavour potential of the genotype. It is suggested that the level of enzyme activities present in unfermented beans is not a limiting factor for optimal formation of flavour precursors during the fermentation process. © 2000 Society of Chemical Industry     

Enzyme activities in cocoa beans during fermentation

The activities of endoprotease, aminopeptidase, carboxypeptidase, invertase (cotyledon and pulp), polyphenol oxidase and glycosidases were studied during heap fermentation of ICS-95 cocoa beans. These enzymes are of key importance in flavour precursor formation and in pigment degradation during cocoa fermentation. Optimal extraction and assay conditions were established to characterise the enzymes reactions and to quantify and compare enzyme activities during cocoa fermentation. The enzymes exhibited large differences in pH optima and stability during fermentation. Aminopeptidase, invertase (cotyledon and pulp) and polyphenol oxidase were strongly inactivated, carboxypeptidase was partly inactivated, whereas endoprotease and glycosidases remained active throughout the fermentation. Since many enzymes are inactivated during fermentation, it is generally recognised that the actual period of enzyme action is short. Although our results confirmed total inactivation for some enzymes, we show that several key enzymes are not completely inactivated during fermentation. Therefore, some enzyme reactions can continue throughout the whole fermentation process. Only polyphenol oxidase was strongly inactivated during sun and artificial drying of the beans. The other enzymes were stable during the drying process. Enzymes like endoprotease and glycosidases are still active in properly fermented and dried beans. © 1998 SCI.     

发酵和焙烤对可可豆多酚、黄酮和风味品质的影响

本实验采用高效液相色谱仪（high performance liquid chromatography,HPLC）、色差仪、电子鼻分别检测未发酵、发酵和焙烤海南可可豆的总酚含量、总黄酮含量、色度和可可豆风味差异。结果表明：未经焙烤的未发酵豆总酚（464.03 mg/10 g）和总黄酮（126.86 mg/10 g）含量明显高于发酵豆总酚（211.86 mg/10 g）和总黄酮（61.98 mg/10 g）含量。105～145 ℃焙烤30 min,未发酵豆总酚和总黄酮含量分别为419.5～129.8 mg/10 g和77.8～16.8 mg/10 g;发酵豆总酚和总黄酮含量分别为182.53～86.25 mg/10 g和34.7～7.0 mg/10 g。其中,125 ℃焙烤20～40 min,未发酵豆总酚和黄酮含量分别为353.74～289.45 mg/10 g和42.86～32.20 mg/10 g;发酵豆总酚和黄酮含量分别为152.08～123.55 mg/10 g和25.12～21.14 mg/10 g。在105～145 ℃的温度焙烤下没食子酸含量变化显著。可可豆色度值的范围：L*值集中在40.0～47.0之间,a*值集中在5.0～6.8之间,b*值集中在4.0～8.5之间。未发酵和发酵可可豆之间,以及不同温度焙烤可可豆之间的电子感官风味分析结果差异较大。     

Free amino acid amounts in raw cocoas from different origins

Raw cocoa is the processed and traded form of the cocoa seed. Fresh seeds undergo fermentation and a drying process before they are prepared for transport and shipping. Depending on the local situation in the producer region the seeds are collected from big estates and are fermented and dried in big lots or they originate from small farmer's crop trees. In the subsequent transport and trading chain raw cocoas may be mixed and stored before they finally are sold and brought to the consumer countries. Local or regional variations in cocoa plant material, fermentation procedures and drying processes finally result in a typical traded good with respect to the amino acids, which form an important part of the flavour precursors. These free amino acids and their composition result from the fermentation procedure of fresh seeds. In the course of the fermentation specific cleavage of cocoa storage proteins delivers the amino acid patterns. In this study the variation of free amino acid amount and distribution of 108 commercially fermented and traded cocoa samples and two Theobroma grandiflorum (Willd. ex Spreng.) K. Schum samples were determined. This examination showed clearly, that content and distribution of free amino acids in raw cocoa from different origins vary greatly (5–25 mg g⁻¹ fatfree dry matter), in some cases country and even region-specific differences were apparent. It is important to notice typical, region-specific variations in the amounts and compositions of free amino acids.     

Fermentation of cocoa beans: influence of microbial activities and polyphenol concentrations on the flavour of chocolate

BACKGROUND: Spontaneous cocoa bean fermentation is characterised by a succession of microbial activities. Cocoa flavour precursors are developed during fermentation and drying of cocoa beans. Polyphenols and alkaloids contribute to astringency and bitterness of cocoa and chocolate. RESULTS: Population dynamics, metabolite target analyses, and chocolate production were performed for seven independent spontaneous cocoa bean heap fermentations in Ghana. Although the same micro‐organisms were involved in these heaps, carried out at different farms or in different seasons, heap temperatures and microbial metabolite concentrations were different. This could be due to heterogeneity and size of the heaps, but was mainly ascribed to microbial variability. Indeed, differences in microbial activity could be linked with the flavour of chocolates made from the corresponding dried, fermented cocoa beans. Whereas the polyphenol and alkaloid contents of cocoa beans were crop‐ and heap‐dependent, epicatechin and theobromine levels decreased during fermentation due to diffusion out of the bean cotyledons and polyphenol oxidation and condensation. Residual levels were responsible for the degree of bitterness of the final chocolates. CONCLUSION: Differences in microbial activities between different heap fermentations can result in dried fermented cocoa beans and chocolates with different flavour characteristics. Hence, fermentation control may direct the flavour of chocolate. Copyright © 2008 Society of Chemical Industry     

Variations of peroxidase activity in cocoa (Theobroma cacao L.) beans during their ripening,fermentation and drying

An increase of peroxidase activity in the seeds of cocoa (Theobroma cacao L.) during their ripening has been determined. An additional increase of the peroxidase activity (about 10 times) is observed during the fermentation and drying of the beans. The residual activity determined in the cocoa beans after sun-drying was higher than that in unfermented ripe seeds. The major cocoa isoperoxidase was shown to be an acidic enzyme with pI 4.7. Using isoelectrofocusing, the appearance of two basic isoenzymes of the peroxidase with pI 8.6 and 9.0 during the process of the fermentation has been detected. ©     

Factors influencing quality variation in cocoa (Theobroma cacao) bean flavour profile — A review

This review examined the factors that influence flavour volatiles of cocoa beans and the volume of work that needs to be done on these factors and their impact on the flavour volatiles of commercial cocoa beans. Cocoa bean flavour is one of the most important quality attributes as flavour is central to acceptability of cocoa beans and cocoa products such as chocolate. The complex composition of cocoa bean flavour depends on bean genotype, postharvest treatments such as pulp pre-conditioning, fermentation and drying, industrial processes such as roasting as well as the type of soil and age of cocoa tree. The bean genotype determines the chemical composition of the bean, specifically the contents of bean storage proteins, polysaccharides, and polyphenols. This determines the quantities and type of precursors formed during fermentation and drying processes leading to flavour formation, hence, influencing both flavour type and intensity. Cocoa bean fermentation and drying result in the breakdown of the storage proteins by endogenous proteases into amino acids and short chain oligopeptides while the polysaccharides are also degraded by invertase to glucose and fructose. The amino acids, oligopeptides, glucose and fructose react with each other during the roasting process to produce the typical cocoa flavour volatiles. Polyphenols are also oxidized by polyphenol oxidase during fermentation and drying which reduce the astringency and bitterness of the beans, thus, enhancing the flavour of cocoa beans. However, the extent to which other factors such as age of the cocoa tree and soil chemical compositions influence the formation of flavour precursors and their relationships with final flavour quality remains unclear. With increasing demand for sustainable production of high quality cocoa beans, greater understanding of factors contributing to the variations in flavour character would have significant commercial implications.     

Performance of different drying methods and their effects on the chemical quality attributes of raw cocoa material

Studies were carried out to investigate the impact of different drying processes on the chemical quality traits of raw cocoa beans. The pH of less fermented cocoa is higher than the well‐fermented cocoa’s. The sun‐dried beans pH ranged from 4.5 to 5.5, while the pH of both oven‐ and mixed‐dried beans was between 3.8 and 5.2. The sun‐dried beans contained lower volatile acidity than oven‐dried beans. Artificially dried beans resulted in higher free acidity content when compared to both sun‐ and mixed‐dried beans. Ammonium nitrogen content in raw cocoa beans is not influenced by the drying methods. Free fatty acid content increases slowly but remains below the critical value of 1.75% whatever the drying processes. While oven‐dried beans show the FFA content above 0.70% both of sun‐ and mixed‐dried beans are associated with FFA content below 0.70%.     

Effect of drying time,bean depth and temperature on free amino acid,peptide‐N,sugar and pyrazine concentrations of Malaysian cocoa beans

The effect of drying time on free amino acid, peptide‐N, sugar and pyrazine concentrations as well as the influence of bean depth and temperature on these compounds during cocoa drying was studied. Drying time, bean depth and temperature significantly decreased the concentration of free amino acids, peptide‐N, total reducing sugars and sucrose and increased the concentration of trimethyl‐, tetramethylpyrazine and total pyrazines in cocoa beans. The best drying treatment was obtained at the combination of bean depth/drying temperature of 8.3 cm/40 °C. This was based on the fact that it produced significantly high concentrations of hydrophobic free amino acids, peptide‐N and total reducing sugars and significantly low concentration of trimethyl‐, tetramethylpyrazine and total pyrazines. Drying treatment of 1.5 cm/60 °C significantly produced the lowest concentration of free amino acids, peptide‐N and total reducing sugars and the highest concentration of pyrazines. © 1999 Society of Chemical Industry     

Effect of farm and industrial processing on the amino acid profile of cocoa beans

An investigation into the amino acid profiles of unfermented and fermented cocoa nibs, as well as process-line cocoa nibs (P-LCN) and processed cocoa cake samples (PCCS) was carried out. In the unfermented cocoa nibs, Glu (128 mg/g crude protein i.e. 128 mg/gcp) was the most abundant amino acid whilst the most concentrated essential amino acid in the same sample was Leu (72.2 mg/g crude protein); in the fermented cocoa nibs, a similar trend was observed, with respective values of Glu (153 mg/gcp) and Leu (62.4 mg/gcp). Lys (181 mg/gcp) was most abundant amino acid in P-LCN and Ile (63.3 mg/gcp) was the second most abundant; also, in PCCS, Lys (52.7 mg/gcp) was the most abundant amino acid but Asp (43.7 mg/gcp) was the second most abundant. The total amino acid content was (mg/gcp), 641 (unfermented nibs), 708 (fermented nibs), 635 (P-LCN) and 368 (PCCS), with corresponding essential amino acids of 300, 287, 478 and 185 (all with His), respectively. Based on whole hen’s egg, the limiting amino acids for the samples were: Ser (unfermented nibs), Met (fermented nibs), Ala (P-LCN) and Val (PCCS), whereas under provisional amino acid scoring pattern, they were: Met + Cys (unfermented nibs), Met + Cys (fermented nibs), Thr (P-LCN) and Val (PCCS). Prolonged and high heat treatments appeared to have reduced the essential amino acids of the PCCS as compared to the P-LCN. Significant differences existed between contents of essential amino acids and non-essential amino acids at p < 0.05 in unfermented cocoa nibs, fermented cocoa nibs and P-LCN.     

Ghanaian Cocoa Bean Fermentation Characterized by Spectroscopic and Chromatographic Methods and Chemometrics

Abstract: Export of cocoa beans is of great economic importance in Ghana and several other tropical countries. Raw cocoa has an astringent, unpleasant taste, and flavor, and has to be fermented, dried, and roasted to obtain the characteristic cocoa flavor and taste. In an attempt to obtain a deeper understanding of the changes in the cocoa beans during fermentation and investigate the possibility of future development of objective methods for assessing the degree of fermentation, a novel combination of methods including cut test, colorimetry, fluorescence spectroscopy, NIR spectroscopy, and GC-MS evaluated by chemometric methods was used to examine cocoa beans sampled at different durations of fermentation and samples representing fully fermented and dried beans from all cocoa growing regions of Ghana. Using colorimetry it was found that samples moved towards higher a* and b* values as fermentation progressed. Furthermore, the degree of fermentation could, in general, be well described by the spectroscopic methods used. In addition, it was possible to link analysis of volatile compounds with predictions of fermentation time. Fermented and dried cocoa beans from the Volta and the Western regions clustered separately in the score plots based on colorimetric, fluorescence, NIR, and GC-MS indicating regional differences in the composition of Ghanaian cocoa beans. The study demonstrates the potential of colorimetry and spectroscopic methods as valuable tools for determining the fermentation degree of cocoa beans. Using GC-MS it was possible to demonstrate the formation of several important aroma compounds such 2-phenylethyl acetate, propionic acid, and acetoin and the breakdown of others like diacetyl during fermentation. Practical Application: The present study demonstrates the potential of using colorimetry and spectroscopic methods as objective methods for determining cocoa bean quality along the processing chain. Development of objective methods for determining cocoa bean quality will be of great importance for quality insurance within the fields of cocoa processing and raw material control in chocolate producing companies.     

发酵牛肉干加工中理化特性与风味品质分析

为探究发酵牛肉干不同加工阶段理化性质和风味品质的变化,以牛臀肉为原料,接种木糖葡萄球菌和清酒乳杆菌制作发酵牛肉干,并对牛肉干在发酵前后及成品的营养成分、质构、色泽等理化特性及游离氨基酸、脂肪酸组成和挥发性风味物质对比分析。结果表明,与发酵前相比,接种复合发酵剂的牛肉干pH值、水分活度、亚硝酸盐残留量、硬度、咀嚼性均显著降低（P＜0.05）;红度值和游离氨基酸含量显著增加,且鲜味氨基酸占比最高;游离脂肪酸以棕榈酸、硬脂酸、油酸、亚油酸为主,其中油酸占比最大;发酵后牛肉检出47 种挥发性化合物,高于发酵前42 种,提高了牛肉干中醛、醇、酚、酯、酸、含氮及其他化合物的种类和相对含量。发酵牛肉干质构及色泽明显改善,游离氨基酸和脂肪酸含量增加,产品营养价值、风味品质及质量安全显著提高。     

Changes in Soya Bean Lipid Profiles during Kinema Production

The influence of processing on soya bean lipids during kinema production was studied. The crude lipid (CL), free fatty acid (FFA) and phytosterol contents of raw beans were 121 g, 6·9 g and 0·8 g kg⁻¹ dry matter, respectively. After soaking and cooking the moisture content of beans increased significantly from 121 to 749 g kg⁻¹, whilst the lipid contents were unchanged. During a 2 day fermentation at 37°C, the lipid contents increased significantly as indicated by capillary column gas chromatography (GC) analysis of petroleum ether extracts of lyophilised samples of unfermented and fermented beans. The fatty acids, palmitic (C16:0), stearic (C18:0), oleic (C18:1), linoleic (C18:2), linolenic (C18:3) and arachidic (C20:0) acids were identified. The major fatty acid present in all samples was the essential fatty acid, linoleic acid. All of the aforementioned fatty acids (except arachidic) were liberated during fermentation by microbial lipases, yielding approximately the same proportion of fatty acids found in unfermented beans. The unsaponifiable soya bean lipids identified by GC were campesterol, stigmasterol and β-sitosterol (ratio 1·2:1:3·4). Levels of these increased during fermentation by 50–61%, but remained in approximately the same proportions.     

Impact of starter cultures and fermentation techniques on the volatile aroma and sensory profile of chocolate

The sensory quality of chocolate is widely determined by the qualitative and quantitative composition of volatile compounds resulting from microbial metabolism during fermentation, and Maillard reactions taking place during drying, roasting and conching. The influence of applying mixed starter cultures on the formation of flavour precursors, composition of volatile aroma compounds and sensory profile was investigated in cocoa inoculated with cultures encompassing a highly aromatic strain of Pichia kluyveri or a pectinolytic strain of Kluyveromyces marxianus, and compared to commercially fermented heap and tray cocoa. Although only minor differences in the concentration of free amino acids and reducing sugars was measured, identification and quantification by dynamic headspace gas chromatography–mass spectrometry (HS/GC–MS) revealed pronounced differences in the composition of volatiles in roasted cocoa liquors and finished chocolates. 19 of the 56 volatile compounds identified in the chocolates were found in significantly higher amounts in the tray fermented sample, whilst significantly higher amounts of 2-methoxyphenol was measured in the two inoculated chocolates. The P. kluyveri inoculated chocolate was characterized by a significantly higher concentration of phenylacetaldehyde and the K. marxianus inoculated chocolate by significantly higher amounts of benzyl alcohol, phenethyl alcohol, benzyl acetate and phenethyl acetate compared to a spontaneously fermented control. Sensory profiling described the heap and tray fermented chocolates as sweet with cocoa and caramel flavours, whilst the inoculated chocolates were characterized as fruity, acid and bitter with berry, yoghurt and balsamic flavours. The choice of fermentation technique had the greatest overall impact on the volatile aroma and sensory profile, but whilst the application of starter cultures did affect the volatile aroma profile, differences were too small to significantly change consumer perception of the chocolates as compared to a spontaneously fermented control.     

Analysis of vicilin (7S)‐class globulin in cocoa cotyledons from various genetic origins

Cocoa cotyledons contain vicilin (7S)‐class globulin (VCG), a major storage protein. It is the native source of oligopeptides and free amino acids which have been identified as precursors of cocoa‐specific aroma and are formed through proteolysis during fermentation. High‐resolution electrophoresis of native proteins isolated from ripe, unfermented cocoa cotyledons harvested from different cultivars was used to determine genetic differences of the genotypes. Flavour differences have been reported to exist after standard fermentation in cocoa beans harvested from various genotypes. In this paper, SDS‐PAGE and 2D IEF/SDS‐PAGE polypeptide separation patterns are shown which were separately isolated from cotyledons of various genetic origins. Cotyledons from three cultivars belonging to genetically distant varieties and a hybrid, Criollo, Forastero and Trinitario, did not reveal any analytical identity differences of VCG subunit polypeptide bands. Additionally, proteins of cotyledons harvested from three of those clones which were reported to produce genotype‐specific flavour differences in raw cocoa after standard fermentation, SCA 12, UIT1 and PBC 140, when analysed in the same way, did not indicate differences. Thus the cotyledon storage proteins from various genetically different cocoa trees are, within methodological limits, the same. We conclude that aroma differences in raw cocoa harvested from various genotypes are the result of other genetic, physiological or curing‐related factors, but are not due to genetic differences of aroma precursors derived from storage proteins. © 2002 Society of Chemical Industry     

Effect of mass and turning time on free amino acid,peptide-N,sugar and pyrazine concentration during cocoa fermentation

A response surface methodology (RSM) was used to determine the optimum condition for mass and turning time during cocoa fermentation. Mass and turning time were used as independent variables; concentrations of free amino acids, peptide-N, sugars and pyrazines were the dependent variables. The R² values for peptide-N, tetramethylpyrazine and total pyrazines were greater than 0·9. Both lower (10 kg) and higher (100 kg) cocoa mass together with lower (0 min) and higher (10 min) turning time gave products containing low concentrations of hydrophobic, total and other free amino acids, peptide-N, fructose, glucose and total reducing sugars; in contrast, those of acidic free amino acids gave higher concentrations. Trimethyl-, tetramethyl- and total pyrazines increased significantly (P<0·05) at higher mass (100 kg) and higher turning time (10 min). From the highest concentration of the important flavour precursors ie hydrophobic free amino acids, total reducing sugars and peptide-N, the recommended mass of cocoa beans and turning time for an optimum cocoa fermentation condition was 60 kg and 5 min, respectively. © 1998 Society of Chemical Industry.     

Acidification,proteolysis and flavour potential in fermenting cocoa beans

Cocoa fermentations in Ghana and Trinidad as well as anaerobic fermentation-like incubations of fresh cocoa beans in Germany were carried out under controlled conditions. Samples of beans were taken during the course of these treatments and determinations were made as to acidification (pH, acetic acid content), proteolysis (free α-amino nitrogen, peptide nitrogen and SDS electrophoresis of the protein peptides) and flavour potential (gas chromatography of the highly volatile compounds, in particular isopentanal and organoleptic analysis after thin layer roasting). A positive correlation between acidification, proteolysis and the development of flavour potential during anaerobic fermentation can be demonstrated in principle. However, the flavour potential is increased if the temperature rise is comparatively slow in both normal fermentation and laboratory incubation. Strong acidification and high accumulation of amino acids and peptides were not essential for a good flavour potential. The isopentanal content proved to be a useful indicator of the progress of normal fermentation in the tropics. These findings can be interpreted on the basis of earlier results about germination-like processes in the protein vacuoles, pre- and post-mortem subcellular structures and the special characteristics of acetic acid diffusion. Conclusions which are relevant to the practice of cocoa fermentation are discussed in more detail.