Methods for fermenting cocoa beans and their uses

Abstract

We provide technology that can control the fermentation of cocoa beans. [Solution] The method for fermenting cocoa beans includes an adjustment step of adjusting the initial pH of the cocoa pulp containing the cocoa beans to between 3.8 and 6.0, and a fermentation step of fermenting the cocoa beans after the adjustment step.

Description

Technical Field

[0001] The present disclosure relates to a method for fermenting cocoa beans.

Background Art

[0002] Cocoa beans are known as raw materials for chocolate and cocoa. Cocoa beans are wrapped in pulp (cocoa pulp) inside the fruit of the cocoa tree (cocoa pod). The cocoa pulp in the state of wrapping the cocoa beans is taken out from the cocoa pod, and then the fermentation of the cocoa beans proceeds by the action of microorganisms. If the fermentation of the cocoa beans does not proceed or the fermentation is not properly controlled, the cocoa beans may become inedible due to spoilage or mold growth, so it is required to promote appropriate fermentation. For example, Patent Document 1 proposes a method for fermenting cocoa beans, which is characterized in that the fermentation of cocoa beans is carried out using a yeast starter.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] The technique described in Patent Document 1 promotes the fermentation of cocoa beans by adding yeast as a starter. However, in the situation where the fermentation of cocoa beans is often carried out in the cocoa production area, there is a problem that it is difficult to control the fermentation, and there is room for improvement from the viewpoint of practical application. For this reason, other techniques capable of controlling the fermentation of cocoa beans are required.

Means for Solving the Problems

[0005] The present disclosure can be realized in the following forms.

[0006] (1) According to one embodiment of the present disclosure, a method for fermenting cocoa beans is provided. This method for fermenting cocoa beans includes an adjustment step of adjusting the initial pH of cocoa pulp containing cocoa beans to 3.8 or more and 6.0 or less, and a fermentation step of fermenting the cocoa beans after the adjustment step. According to this embodiment of the method for fermenting cocoa beans, the fermentation of cocoa beans can be controlled.

[0007] (2) In the cocoa bean fermentation method described in (1) above, the initial pH of the cocoa pulp may be adjusted to 4.0 or higher and 5.5 or lower in the adjustment step. This form of cocoa bean fermentation method allows for more effective control of the cocoa bean fermentation.

[0008] (3) In the cocoa bean fermentation method described in (1) or (2) above, the pH may be adjusted in the adjustment step using at least one selected from the group consisting of NaHCO3, CaCO3, and a phosphate buffer. This form of cocoa bean fermentation method allows for easy control of the fermentation of cocoa beans.

[0009] (4) In the cocoa bean fermentation method described in any one of the above items (1) to (3), the pH may be adjusted using NaHCO3 in the adjustment step. This form of cocoa bean fermentation method can suppress the occurrence of problems due to components remaining in the fermented cocoa beans.

[0010] (5) In the cocoa bean fermentation method described in any one of the above items (1) to (4), in the preparation step, 0.25% by mass or more and 1.5% by mass or less of NaHCO3 may be mixed with the mass of the cocoa pulp. This form of cocoa bean fermentation method allows for more effective control of the cocoa bean fermentation.

[0011] (6) In the method for fermenting cocoa beans described in any one of the above items (1) to (5), the preparation step further comprises adding at least one microorganism selected from the group consisting of lactic acid bacteria, yeast, and acetic acid bacteria, in an amount of 10% relative to the mass of the cocoa pulp. 5 CFU / g or more (10) 9The mixture may be mixed to a CFU / g or less. This method of fermenting cocoa beans allows for more effective control of the fermentation process.

[0012] (7) In the method for fermenting cocoa beans described in (6) above, the microorganisms may include lactic acid bacteria. This form of method for fermenting cocoa beans can further promote lactic acid fermentation, and thus the fermentation of cocoa beans can be controlled more effectively.

[0013] (8) In the method for fermenting cocoa beans described in (6) or (7) above, the microorganism may include acetic acid bacteria. According to this form of method for fermenting cocoa beans, the amount of malic acid produced can be increased by mixing in acetic acid bacteria. As a result, the pH in the fermentation process can be lowered, and the aroma of the fermented cocoa beans can be improved.

[0014] (9) In the cocoa bean fermentation method described in any one of the above paragraphs (6) to (8), the adjustment step may involve adjusting the initial pH of the cocoa pulp before mixing in the microorganisms. This form of cocoa bean fermentation method makes it possible to suppress the influence of the mixed microorganisms on the pH adjustment process.

[0015] (10) Another embodiment of the present disclosure provides a method for producing fermented cocoa beans. This method for producing fermented cocoa beans includes an adjustment step of adjusting the initial pH of cocoa pulp containing cocoa beans to 3.8 or higher and 6.0 or lower, and a fermentation step of fermenting the cocoa beans after the adjustment step. This embodiment of the method for producing fermented cocoa beans allows for control of the fermentation of cocoa beans, thereby suppressing a decrease in the production efficiency of fermented cocoa beans.

[0016] (11) Another embodiment of the present disclosure provides a method for producing cocoa nibs, which includes a roasting step of roasting fermented cocoa beans obtained by the manufacturing method described in (10) above. This embodiment of the method for producing cocoa nibs allows for control of the fermentation of the cocoa beans, thereby suppressing a decrease in the efficiency of cocoa nib production.

[0017] (12) According to another aspect of the present disclosure, there is provided a method for producing cocoa mass, including a grinding step of grinding the cocoa nibs obtained by the production method described in (11) above. According to the method for producing cocoa mass of this aspect, since the fermentation of cocoa beans can be controlled, it is possible to suppress a decrease in the production efficiency of cocoa mass.

[0018] (13) According to another aspect of the present disclosure, there is provided a method for producing cocoa butter or cocoa powder, including a pressing step of pressing the cocoa mass obtained by the production method described in (12) above. According to the method for producing cocoa butter or cocoa powder of this aspect, since the fermentation of cocoa beans can be controlled, it is possible to suppress a decrease in the production efficiency of cocoa butter or cocoa powder.

[0019] (14) According to another aspect of the present disclosure, there is provided a method for producing chocolate, including a mixing step of mixing cocoa butter into the cocoa mass obtained by the production method described in (12) above. According to the method for producing chocolate of this aspect, since the fermentation of cocoa beans can be controlled, it is possible to suppress a decrease in the production efficiency of chocolate.

[0020] Note that the present disclosure can be realized in various forms. For example, it can be realized in forms such as a method for producing a cocoa mass blended product including a step of blending the cocoa mass obtained by the above production method, a method for producing a cocoa blended product including a step of blending the cocoa obtained by the above production method, and a method for producing a chocolate blended product including a step of blending the chocolate obtained by the above production method.

Brief Description of the Drawings

[0021] [Figure 1] It is an explanatory diagram showing the relationship between the amount of NaHCO3 added and the initial pH. [Figure 2] It is an explanatory diagram showing the relationship between the amount of NaHCO3 added and the pH during the fermentation process. [Figure 3]It is an explanatory diagram showing the relationship between the amount of NaHCO3 added and the amount of lactic acid produced during the fermentation process. [Figure 4] It is an explanatory diagram showing the HPLC analysis result of the blank. [Figure 5] It is an explanatory diagram showing the HPLC analysis result of the sample inoculated with acetic acid bacteria. [Figure 6] It is an explanatory diagram showing an enlarged part of FIG. 5.

Embodiments for Carrying Out the Invention

[0022] [[ID=**16]] Generally, the fermentation of cacao beans is often carried out in the cacao-producing areas. The fermentation of cacao beans first involves harvesting cacao pods from the log of Theobroma cacao L., and then removing the cacao pulp that wraps the cacao beans from the cacao pods, and the fermentation is carried out by depositing the cacao pulp. Examples of fermentation methods include the heap method and the box method. In the heap method, the cacao pulp is piled up and covered with banana leaves or sheets to allow the fermentation to proceed. In the box method, the cacao pulp is placed in a wooden box or the like for fermentation to proceed. Multiple types of microorganisms act on the fermentation. In the initial stage of fermentation, the fermentation proceeds in an anaerobic environment with little oxygen. At this stage, yeasts and lactic acid bacteria grow, and the sugars in the cacao pulp are converted into ethanol by the action of yeasts and into lactic acid by the action of lactic acid bacteria. Then, by performing stirring and mixing, oxygen is supplied to the cacao beans and the environment switches to an aerobic one. At this stage, acetic acid bacteria grow, and the ethanol produced by yeasts is converted into acetic acid by the action of acetic acid bacteria, and the lactic acid produced by lactic acid bacteria is converted into other organic acids. The microorganisms that contributed to the fermentation die and decrease in the later stage of fermentation due to the metabolites they produced, the rise in temperature, etc. And general bacteria such as spore bacteria slowly grow. The metabolites such as alcohols and organic acids produced along with such changes in microorganisms have a great influence on the quality of cacao beans and the flavor of chocolate and the like.

[0023] It should be noted that there may be some inaccuracies in the translation due to the complexity of the technical content. It is recommended to consult relevant professionals for more accurate translation in actual patent translation work.According to one embodiment of the present disclosure, a method for fermenting cocoa beans is provided. This method for fermenting cocoa beans includes an adjustment step of adjusting the initial pH of cocoa pulp containing cocoa beans to between 3.8 and 6.0, and a fermentation step of fermenting the cocoa beans after the adjustment step. In the present disclosure, "initial pH of cocoa pulp" means the pH of cocoa pulp removed from cocoa pods before being subjected to fermentation by the heap method, box method, or the like. The initial pH of cocoa pulp is measured by bringing a pH electrode into contact with the cocoa pulp.

[0024] In the preparation process, the initial pH of the cocoa pulp is adjusted to 6.0 or lower to prevent excessive promotion of fermentation by lactic acid bacteria during the fermentation process. This suppresses over-fermentation of cocoa beans, thereby preventing the fermented cocoa beans and products using them as raw materials from becoming too acidic. Furthermore, adjusting the initial pH of the cocoa pulp to 6.0 or lower prevents the enzymes of the microorganisms contributing to fermentation from deviating from the optimal pH, thus preventing fermentation from stalling. Additionally, in the preparation process, the initial pH of the cocoa pulp is adjusted to 3.8 or higher to promote fermentation by lactic acid bacteria during the fermentation process. In the preparation process, from the viewpoint of more effectively controlling the fermentation of cocoa beans, it is preferable to adjust the initial pH of the cocoa pulp between 4.0 and 6.0, and more preferably between 4.0 and 5.5.

[0025] In the adjustment process, the pH can be adjusted by mixing an alkaline substance or a buffering agent that has a buffering effect in the weakly acidic to neutral range with the cocoa pulp. Examples of alkaline substances include NaHCO3 (sodium bicarbonate), CaCO3 (calcium carbonate), K2CO3 (potassium carbonate), and Ca(OH)2 (calcium hydroxide). Examples of buffering agents include phosphate buffers. In the adjustment process, from the viewpoint of easily controlling the fermentation of cocoa beans, it is preferable to adjust the pH using at least one selected from the group consisting of NaHCO3, CaCO3, and phosphate buffers. From the viewpoint of suppressing problems caused by components remaining in the fermented cocoa beans, it is more preferable to adjust the pH using NaHCO3.

[0026] When NaHCO3 is used in the preparation process, from the viewpoint of effectively controlling the fermentation of cocoa beans, it is preferable to mix 0.25% to 1.5% by mass of NaHCO3 with respect to the mass of cocoa pulp, and more preferably 0.3% to 1.3% by mass of NaHCO3. In this disclosure, "mass of cocoa pulp" does not include the mass of cocoa beans inside the cocoa pulp.

[0027] In the preparation process, at least one microorganism selected from the group consisting of lactic acid bacteria, yeast, and acetic acid bacteria is added in a quantity of 10% relative to the mass of the cocoa pulp. 5 CFU / g or more (10) 9 It is preferable to mix the cocoa beans so that the CFU / g is 100% or less. By mixing in microorganisms that contribute to fermentation, the time required for microbial growth can be shortened. As a result, the fermentation of cocoa beans can be accelerated, and thus the fermentation of cocoa beans can be controlled more effectively. From the viewpoint of further promoting lactic acid fermentation and controlling the fermentation of cocoa beans more effectively, it is preferable that the microorganisms mixed in the preparation step include lactic acid bacteria. In other words, in the preparation step, microorganisms containing lactic acid bacteria should be mixed in an amount of 100% relative to the mass of cocoa pulp. 5 CFU / g or more (10) 9It is preferable to mix the microorganisms so that the CFU / g is 10% or less. Furthermore, it is preferable that the microorganisms mixed in the preparation step include acetic acid bacteria. In other words, in the preparation step, the microorganisms containing acetic acid bacteria are mixed in an amount equal to 10% of the mass of cocoa pulp. 5 CFU / g or more (10) 9 It is preferable to mix the ingredients so that the CFU / g level is 30% or less. As shown in the examples described later, the amount of malic acid produced can be increased by adding acetic acid bacteria. As a result, the pH in the fermentation process can be lowered. Lowering the pH suppresses the germination of cocoa beans and promotes autodigestion of cocoa beans, thereby suppressing the occurrence of bitterness and astringency while increasing the amount of precursors that contribute to flavor. In addition, increasing the amount of malic acid increases the amount of precursors that contribute to flavor, thereby improving the aroma of fermented cocoa beans. As a result, the aroma of cocoa nibs, cocoa mass, cocoa butter, cocoa powder, chocolate, etc. produced using fermented cocoa beans can be improved. The microorganisms mixed in the preparation process may be one type or a combination of two or more types.

[0028] The lactic acid bacteria mixed in the preparation process are not particularly limited, but examples include Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus collinoides, Lactobacillus mali, Lactobacillus casei, Lactococcus lactis, Leuconostoc mesenteroides, Pediococcus pentosaceus, Streptococcus thermophilus, and Oenococcus oeni. However, it is preferable to include at least one of Lactobacillus rhamnosus and Leuconostoc mesenteroides. The acetic acid bacteria mixed in the preparation process are not particularly limited, but examples include Acetobacter aceti, Acetobacter pasteurianus, Acetobacter ghanensis, Acetobacter ascendens, Acetobacter rancens, Acetobacter xylinum, and Glucononbacter oxydans, but it is preferable that they include Acetobacter aceti.The yeasts mixed in the preparation process are not particularly limited, but examples include Saccharomyces spp. such as Saccharomyces serevusiae, Kluyveromyces spp. such as Kluyveromyces marxianus and Kluyveromyces lactis, Candida spp. such as Candida krusei and Candida zempinina, Galactomyces spp. such as Galactomyces geotrichum, Wickerhamomyces spp. such as Wickerhamomyces anomalus, Torulaspora spp. such as Torulaspora delbrueckii, Pichia spp. such as Pichia kudriavzevii and Pichia membranafaciens, Debaryomyces spp. such as Debaryomyces hansenii, Hanseniaspora spp., Kloeckera spp., and Saccharomycopsis. Examples include Schizosaccharomyces spp., Schizosaccharomyces spp., Torulopsis spp., etc., but it is preferable to include Saccharomyces serevusiae.

[0029] When microorganisms are added to cocoa pulp during the preparation process, the microorganisms may be mixed after adjusting the initial pH of the cocoa pulp, or the initial pH of the cocoa pulp may be adjusted after mixing the microorganisms, or the mixing of microorganisms and pH adjustment may be performed simultaneously. In the preparation process, it is preferable to adjust the initial pH of the cocoa pulp before mixing the microorganisms, from the viewpoint of suppressing the influence of the pH adjustment work on the microorganisms being mixed.

[0030] The fermentation process may be carried out under static conditions or under stirring conditions after the preparation process, but it is preferable that the entire mixture be stirred at least once between 24 and 96 hours after the completion of the preparation process to switch from anaerobic to aerobic conditions. Stirring is also preferable from the viewpoint of suppressing uneven fermentation. The conditions for fermentation in the fermentation process are not particularly limited, but for example, the fermentation may be carried out under conditions such as a fermentation temperature of 10°C to 50°C, preferably 20°C to 45°C, a fermentation time of 1 day to 10 days, preferably 2 days to 7 days, more preferably 3 days to 5 days. In addition, other arbitrary substances such as fruit pulp or juice may be added during the fermentation process.

[0031] In the fermentation process, the amount of lactic acid produced (g) relative to the mass (kg) of cocoa pulp is preferably 8 g / kg to 32 g / kg, preferably 13 g / kg to 30 g / kg, and more preferably 18 g / kg to 28 g / kg. The amount of lactic acid produced can be measured by centrifugation of the fermented product (15,000 rpm, 4°C, 5 minutes), collection of the supernatant, and then appropriate dilution with distilled water before measurement by high-performance liquid chromatography (HPLC). The measurement conditions for HPLC can be those described in A Ohnishi et.al., RSC Adv., 2012, 2, 8332-8340 (DOI: 10.1039 / C2RA20590D).

[0032] The cocoa bean fermentation method disclosed herein allows for control of cocoa bean fermentation. Furthermore, because the fermentation of cocoa beans can be controlled by a simple method, it can be easily controlled even when fermentation takes place in the cocoa-producing region. In addition, the cocoa bean fermentation method disclosed herein allows for the progression of cocoa bean fermentation, thereby suppressing fermentation failures. As a result, spoilage and mold growth can be suppressed, thus preventing a decline in the quality of fermented cocoa beans. Moreover, the cocoa bean fermentation method disclosed herein prevents excessive fermentation, thus preventing a decline in the quality of fermented cocoa beans. As a result, a decline in the flavor of fermented cocoa beans and products made from fermented cocoa beans can be prevented, thereby improving their palatability.

[0033] Another form of this disclosure provides a method for producing fermented cocoa beans. This method includes an adjustment step of adjusting the initial pH of cocoa pulp containing cocoa beans to 3.8 or higher and 6.0 or lower, and a fermentation step of fermenting the cocoa beans after the adjustment step. The adjustment step and the fermentation step can each be carried out in the same manner as the cocoa bean fermentation method described above. From the viewpoint of suppressing mold growth during storage and transportation, the method for producing fermented cocoa beans of this disclosure preferably includes a drying step after the fermentation step in which the cocoa beans are dried to reduce their moisture content. The drying step is not particularly limited, but may be, for example, sun drying or hot air drying. The method for producing fermented cocoa beans of this disclosure allows for control of the fermentation of cocoa beans, thereby suppressing a decrease in the production efficiency of fermented cocoa beans.

[0034] Another form of this disclosure provides a method for producing cocoa nibs. This method for producing cocoa nibs includes a roasting step of roasting fermented cocoa beans obtained by the method for producing fermented cocoa beans of this disclosure. The roasting step is not particularly limited, and the beans may be roasted after removing the shell and germ from the fermented cocoa beans, or the fermented cocoa beans may be roasted as they are. The roasting conditions are not particularly limited, and for example, the roasting may be carried out at a temperature of 90°C to 140°C and a roasting time of 30 minutes to 120 minutes. The method for producing cocoa nibs of this disclosure allows for control of the fermentation of cocoa beans, thereby suppressing a decrease in the efficiency of cocoa nib production.

[0035] Another form of the present disclosure provides a method for producing cocoa mass. This method for producing cocoa mass includes a grinding step of grinding cocoa nibs obtained by the method for producing cocoa nibs of the present disclosure. The grinding step is not particularly limited, but could include, for example, a method of making a paste using a grinder. The method for producing cocoa mass of the present disclosure allows for control of the fermentation of cocoa beans, thereby suppressing a decrease in the efficiency of cocoa mass production.

[0036] Other forms of this disclosure provide a method for producing cocoa butter or cocoa powder. This method for producing cocoa butter or cocoa powder includes a pressing step of pressing the cocoa mass obtained by the method for producing cocoa mass of this disclosure. The pressing step is not particularly limited, but could include, for example, a method of extracting the fats and oils using a press. Cocoa butter is obtained by refining these fats and oils as needed. Cocoa powder is obtained by grinding the solids (cocoa cake) from which the fats and oils have been removed. The method for producing cocoa butter or cocoa powder of this disclosure allows for control of the fermentation of cocoa beans, thereby suppressing a decrease in the efficiency of producing cocoa butter or cocoa powder.

[0037] A method for producing chocolate is provided in other forms of this disclosure. This method for producing chocolate includes a mixing step of mixing cocoa butter with cocoa mass obtained by the method for producing cocoa mass of this disclosure. The mixing step is not particularly limited and may include sugar and milk as needed. The method for producing chocolate of this disclosure allows for control of the fermentation of cocoa beans, thereby suppressing a decrease in the efficiency of chocolate production. [Examples]

[0038] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0039] <Experiment 1> Confirmation of pH by adding a pH adjuster The pH of cocoa pulp after adding a pH adjuster was determined. First, cocoa pulp was extracted from harvested cocoa pods. 0.1 g of NaHCO3 was added to 20 g of cocoa pulp. After stirring for 5 minutes, the pH was measured by contacting the cocoa pulp with a pH electrode (manufactured by HORIBA). The addition of NaHCO3, stirring, and pH measurement were repeated until the total amount of added NaHCO3 reached 0.6 g. The pH of cocoa pulp without added NaHCO3 was also measured in the same manner. The sample size (n) in Experiment 1 was 2.

[0040] Figure 1 is an explanatory diagram showing the relationship between the amount of NaHCO3 added and the initial pH. In Figure 1, the horizontal axis represents the amount of NaHCO3 added (g), and the vertical axis represents pH. From the results shown in Figure 1, the following was found: The pH of cocoa pulp could be increased by adding NaHCO3, and the pH increased as the amount added increased. More specifically, the pH of cocoa pulp without added NaHCO3 was approximately 3.5, and the pH increased by approximately 1.0 for every 0.1g of NaHCO3 added per 20g of cocoa pulp. This effect of NaHCO3 addition on pH occurred in a manner dependent on the amount added, up to approximately pH 8.0. Therefore, it was shown that the initial pH of cocoa pulp fermentation can be easily and appropriately controlled by adding NaHCO3.

[0041] <Experiment 2> Confirmation of lactic acid production by adding pH adjuster The amount of lactic acid produced when fermentation was carried out with the addition of a pH adjuster was confirmed. For fermentation, the test strain of lactic acid bacteria (Lacticaseibacillus rhamnosus) was added. As the test medium, MRS (de Man Rogosa Sharpe) synthetic medium with the composition shown in Table 1 below was used. Note that all values ​​in Table 1 represent mass percent. For pre-culturing, the test strain was inoculated into the test medium and cultured anaerobically for 24 to 48 hours. The anaerobic environment was maintained using Aneropack (registered trademark, manufactured by Mitsubishi Gas Corporation). After culturing, the grown bacterial cells were collected in sterile water and a cell-detached suspension was prepared.

[0042] [Table 1]

[0043] To 20 g of cocoa pulp, 0-0.6 g of NaHCO3 was added and stirred for 5 minutes. The initial pH was then measured by contacting the cocoa pulp with a pH electrode (HORIBA). Subsequently, a suspension of cells detached from the test strain was inoculated into the cocoa pulp so that the OD660 ≈ 0.1, and cultured at 37°C. Stirring was performed on the third day of culture. pH and lactic acid production were measured over time during the fermentation process. pH was measured by contacting the cocoa pulp with a pH electrode (HORIBA). Lactic acid production was measured by HPLC. The fermented product was centrifuged (15000 rpm, 4°C, 5 minutes), the supernatant was collected, filtered through a glass fiber filter (Whatman GF / C) (Whatman is a registered trademark), and then diluted appropriately with distilled water before being subjected to HPLC. HPLC was performed according to the method described in A Ohnishi et.al., RSC Adv., 2012, 2, 8332-8340 (DOI: 10.1039 / C2RA20590D). More specifically, measurements were performed using an HPLC (PU-2080 Intelligent pump, JASCO), a Gelpack column (GL-C610HS, particle size 6 μm, inner diameter 7.8 mm, length 300 mm, Hitachi Chemical Co., Ltd.), a post-column reactor (RU 2080-51 reaction coil unit, JASCO), an intelligent UV / VIS detector (UV-2070, JASCO), and a data processor (Chromatograph 21, SIC). 3 mM HClO4 (flow rate 0.5 mL / min) was used as the mobile phase. Lactic acid was detected by a post-labeling method using 0.2 mM bromothymol blue and 15 mM Na2HPO4 (flow rate 1.5 mL / min). The column temperature was 60°C and the detection wavelength was 445 nm.

[0044] Figure 2 is an explanatory diagram showing the relationship between the amount of NaHCO3 added and the pH during the fermentation process. In Figure 2, the horizontal axis represents the culture time (hr), the vertical axis represents the pH, and the values ​​in each legend represent the amount of NaHCO3 added (g). From the results shown in Figure 2, the following was found: In samples to which 0.05g to 0.5g of NaHCO3 was added to 20g of cocoa pulp, the initial pH, which increased in proportion to the amount of NaHCO3 added, decreased significantly by the third day of culture. After that, a tendency for the pH to gradually decrease as the culture time increased was observed. This is thought to be due to organic acids such as lactic acid and acetic acid produced by fermentation. In addition, in samples to which NaHCO3 was not added, the initial pH was low, so the change in pH during culture was small. This is thought to be due to the slow progress of fermentation and the small amount of lactic acid and acetic acid produced. Furthermore, in a sample where 0.6g of NaHCO3 was added to 20g of cocoa pulp, the initial pH was around 7.0, and the decrease in pH was small even with prolonged incubation time. This is thought to be because the buffering effect of the added NaHCO3 kept the pH at a high value, remaining outside the optimal pH for the lactic acid bacteria enzymes, thus hindering lactic acid fermentation.

[0045] Figure 3 is an explanatory diagram showing the relationship between the amount of NaHCO3 added and the amount of lactic acid produced during the fermentation process. In Figure 3, the horizontal axis represents the culture time (hr), the vertical axis represents the amount of lactic acid produced (g / kg of cocoa pulp), and the numerical values ​​in each legend represent the amount of NaHCO3 added (g). From the results shown in Figure 3, the following was found: In samples to which 0.05g to 0.5g of NaHCO3 was added to 20g of cocoa pulp, the amount of lactic acid produced was higher compared to samples to which NaHCO3 was not added, and the amount of lactic acid produced tended to increase with higher initial pH. Furthermore, in samples to which 0.6g of NaHCO3 was added to 20g of cocoa pulp, the amount of lactic acid produced was lower than in samples to which NaHCO3 was not added.

[0046] As shown in Figures 2 and 3, it was found that the amount of lactic acid produced can be controlled by adjusting the initial pH of the cocoa pulp. Therefore, it is thought that by adjusting the initial pH of the cocoa pulp, it is possible to prevent poor fermentation and spoilage of cocoa beans, as well as to control the fermentation of cocoa beans to the desired level.

[0047] <Experiment 3> Fermentation test by adding pH adjuster and inoculating with acetic acid bacteria A fermentation test was conducted by adding a pH adjuster and inoculating with acetic acid bacteria. The acetic acid bacteria strain used was Acetobacter pasteurianus TK4-OL. The culture medium used had the composition shown in Table 2 below. Note that all values ​​in Table 2 represent mass percent. For pre-culturing, the culture medium (agar medium) was inoculated with the culture strain and cultured for 24 to 48 hours. After culturing, the grown bacterial cells were collected in sterile water and a cell-detached suspension was prepared.

[0048] [Table 2]

[0049] 100g of cocoa pulp was placed in a beaker, and NaHCO3 was added to adjust the initial pH to 5.5. Then, 10g portions were dispensed into test tubes. pH was measured by contacting a pH electrode (HORIBA) with the cocoa pulp. The pH of the cocoa pulp before NaHCO3 addition was approximately 3.5. Subsequently, a suspension of cells detached from the test strain was inoculated into the cocoa pulp to an OD660 ≈ 0.1, and cultured at 37°C. As a blank, fermentation of cocoa pulp was carried out under the same conditions except that acetic acid bacteria were not added. On days 4, 8, 12, 16, and 20 of fermentation, the test tubes were stirred, and pH measurement and organic acid analysis were performed. pH was measured by contacting a pH electrode (HORIBA) with the cocoa pulp. Organic acid analysis was performed by HPLC. The fermented product was centrifuged (15,000 rpm, 4°C, 5 minutes), and the supernatant was collected. This supernatant was then filtered through a glass fiber filter (Whatman GF / C) (Whatman is a registered trademark) and subjected to undiluted HPLC. HPLC was performed according to the method described in A Ohnishi et.al., RSC Adv., 2012, 2, 8332-8340 (DOI: 10.1039 / C2RA20590D). More specifically, the measurements were performed using an HPLC (PU-2080 Intelligent pump, JASCO), a Gelpack column (GL-C610HS, particle size 6 μm, inner diameter 7.8 mm, length 300 mm, Hitachi Chemical Co., Ltd.), a post-column reactor (RU 2080-51 reaction coil unit, JASCO), an intelligent UV / VIS detector (UV-2070, JASCO), and a data processor (Chromatocorder 21, SIC). 3 mM HClO4 (flow rate 0.5 mL / min) was used as the mobile phase. The column temperature was 60°C, and the detection wavelength was 445 nm.

[0050] The pH measurement results are shown in Table 3 below. According to the results in Table 3, it was found that the pH decreased further over time after inoculation with acetic acid bacteria. This suggests the possibility that acetic acid fermentation is occurring.

[0051] [Table 3]

[0052] Figure 4 is an explanatory diagram showing the HPLC analysis results of the blank sample. In Figure 4, the peak positions of citric acid, malic acid, succinic acid, lactic acid, and acetic acid are indicated by arrows. As shown in Figure 4, in the blank sample, lactic acid increased and malic acid decreased on day 16 of fermentation.

[0053] Figure 5 is an explanatory diagram showing the results of HPLC analysis of a sample inoculated with acetic acid bacteria. Figure 6 is an enlarged explanatory diagram of a part of Figure 5. In Figure 5, the peak positions of citric acid, malic acid, and succinic acid are indicated by arrows, and in Figure 6, the peak positions of lactic acid and acetic acid are indicated by arrows. In addition, the spectra of the standard substances used in the calibration curve are also shown in Figures 5 and 6. As shown in Figure 5, in the sample inoculated with acetic acid bacteria, malic acid increased significantly with the progression of fermentation time. Also, as shown in Figure 6, a tendency for lactic acid and acetic acid to decrease with the progression of fermentation time was observed. This indicates that lactic acid and acetic acid were consumed by metabolism.

[0054] Table 4 below summarizes the HPLC analysis results for the blank and acetic acid bacteria inoculated samples. Since lactic acid and acetic acid were measured undiluted in this experiment, quantitative results are shown for lactic acid and acetic acid, while the qualitative results for citric acid, malic acid, and succinic acid indicate the degree of content. In Table 4, "◎" indicates a relatively high content, "○" indicates a moderate content, and "△" indicates a relatively low content. Shaded areas in Table 4 indicate that the analysis results were outside the calibration curve. As shown in Table 4, the production of malic acid increased significantly upon inoculation with acetic acid bacteria. The decrease in pH shown in Table 3 is also considered to be due to the increase in malic acid.

[0055] [Table 4]

[0056] The present invention is not limited to the embodiments described above, and can be realized in various configurations without departing from its spirit. For example, the technical features in the embodiments and examples corresponding to the technical features in each form described in the summary of the invention can be replaced or combined as appropriate in order to solve some or all of the above-described problems, or to achieve some or all of the above-described effects. Furthermore, if a technical feature is not described as essential in this specification, it can be deleted as appropriate.

Claims

1. A method of fermenting cocoa beans, The process involves adjusting the initial pH of the cocoa pulp containing the cocoa beans to between 3.8 and 6.0, Following the preparation step, a fermentation step is performed in which the cocoa beans are fermented. A method of fermenting cocoa beans, including [specific ingredient / method].

2. In the method for fermenting cocoa beans according to claim 1, In the aforementioned adjustment step, the initial pH of the cocoa pulp is adjusted to between 4.0 and 5.

5. The fermentation method for cocoa beans.

3. In the method for fermenting cocoa beans according to claim 1, In the aforementioned adjustment step, NaHCO ３ And, CaCO ３ The pH is adjusted using at least one selected from the group consisting of a phosphate buffer and a phosphate buffer. The fermentation method for cocoa beans.

4. In the method for fermenting cocoa beans according to claim 1, In the aforementioned adjustment step, NaHCO ３ Use to adjust the pH. The fermentation method for cocoa beans.

5. In the method for fermenting cocoa beans according to claim 4, In the aforementioned preparation step, 0.25% by mass or more and 1.5% by mass or less of NaHCO3 is added relative to the mass of the cocoa pulp. ３ Mix them together. The fermentation method for cocoa beans.

6. In the method for fermenting cocoa beans according to claim 1, In the aforementioned preparation step, at least one microorganism selected from the group consisting of lactic acid bacteria, yeast, and acetic acid bacteria is added in an amount of 10% relative to the mass of the cocoa pulp. ５ CFU / g or more: 10 ９ Mix so that the CFU / g level is 30% or less. The fermentation method for cocoa beans.

7. In the method for fermenting cocoa beans according to claim 6, The aforementioned microorganisms include lactic acid bacteria, The method of fermenting cocoa beans.

8. In the method for fermenting cocoa beans according to claim 6, The aforementioned microorganisms include acetic acid bacteria, The fermentation method for cocoa beans.

9. In the method for fermenting cocoa beans according to claim 6, In the aforementioned preparation step, the initial pH of the cocoa pulp is adjusted, and then the microorganisms are mixed in. The method of fermenting cocoa beans.

10. A method for producing fermented cocoa beans, A preparation process to adjust the initial pH of the cocoa pulp containing the cocoa beans to between 3.8 and 6.0, Following the preparation step, a fermentation step is performed in which the cocoa beans are fermented. A method for producing fermented cocoa beans, including [the specified ingredient].

11. The method includes a roasting step of roasting the fermented cocoa beans obtained by the manufacturing method described in claim 10, A method for producing cocoa nibs.

12. The method includes a grinding step of grinding the cocoa nibs obtained by the manufacturing method described in claim 11, A method for producing cocoa mass.

13. The method includes a pressing step of pressing the cocoa mass obtained by the manufacturing method described in claim 12, A method for producing cocoa butter or cocoa powder.

14. The method includes a mixing step of mixing cocoa butter with cocoa mass obtained by the manufacturing method described in claim 12. Chocolate manufacturing method.

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