Enzyme preparation, processed coffee and use thereof

Abstract

An object of the present application is to provide a coffee beverage in which coagulation of coffee oil is inhibited. The present application relates to an enzyme preparation for inhibiting coagulation of coffee oil, comprising a lipase from the genus Candida. In addition, the present application relates to a manufacturing method of a processed coffee, comprising subjecting an enzyme preparation to coffee.

Description

Technical Field

[0001] This invention relates to enzyme preparations, processed coffee, methods for manufacturing processed coffee, and methods for inhibiting the aggregation of coffee oil. Background Technology

[0002] The consumption of coffee beverages has been increasing year by year, especially in the Chinese market, which has experienced explosive growth due to rising national income and changing consumer awareness. In the coffee beverage manufacturing process, the extracted coffee undergoes filtration and sterilization.

[0003] In the extraction and filtration processes of coffee beverage manufacturing, various enzymes have been studied for adding to increase the extract content or improve filterability. For example, mannanase is known to act on galactomannan contained in coffee to increase the extraction rate of coffee components and prevent precipitation. Additionally, cellulase is known to act on cellulose, the main component of the cell wall of coffee beans, to increase the solubility of coffee beans, thereby increasing the extraction rate and preventing precipitation.

[0004] In addition, there have been studies on adding enzymes to enhance the aroma or flavor of coffee during the coffee beverage manufacturing process. For example, Patent Document 1 discloses a method for producing coffee flavor by treating coffee oil with lipase. This patent document studies how to produce coffee-flavored products with a pleasant tongue texture, throat feel, and richness by acting lipase on oil derived from coffee beans.

[0005] Existing technical documents

[0006] Patent documents

[0007] Patent Document 1: Japanese Patent Application Publication No. 2007-61046 Summary of the Invention

[0008] Because coffee beans contain oils, during the preservation of coffee extracted from coffee beans, the oils continuously rise to the surface of the coffee and condense (phase separation), sometimes forming oil rings.

[0009] Therefore, the objective of this invention is to provide a coffee beverage that can inhibit the coagulation of coffee oil.

[0010] The following are examples of specific embodiments of the present invention.

[0011] [1] An enzyme preparation for inhibiting the aggregation of coffee oil, comprising a lipase from the genus Candida.

[0012] [2] A method of processing coffee by applying the enzyme preparation described in [1] to coffee.

[0013] [3] The coffee is processed according to [2], wherein the moisture content of the coffee is 80% by mass or more.

[0014] [4] A method for processing coffee, comprising acting a lipase from Candida spp. on coffee.

[0015] [5] The coffee processing method according to [4] wherein the moisture content of the coffee is 80% by mass or more.

[0016] [6] A method for inhibiting the aggregation of coffee oil includes acting a lipase from the genus Candida on coffee.

[0017] According to the present invention, a coffee beverage capable of inhibiting the aggregation of coffee oil can be provided. Furthermore, according to the present invention, a coffee beverage with excellent preservation stability can be provided by inhibiting the aggregation of coffee oil. Detailed Implementation

[0018] The present invention will now be described in detail. The descriptions herein are sometimes based on representative embodiments or specific examples, but the present invention is not limited to such embodiments. It should be noted that the numerical range indicated by “~” in this specification refers to the range including the values ​​described before and after “~” as lower and upper limits.

[0019] (Enzyme preparations)

[0020] This invention relates to an enzyme preparation containing lipase for inhibiting the aggregation of coffee oil. A preferred embodiment of the invention relates to an enzyme preparation containing lipase from the genus *Candida* for inhibiting the aggregation of coffee oil. In this embodiment, by acting the lipase from *Candida* on the coffee, the aggregation of coffee oil can be inhibited, thereby inhibiting the formation of oil rings. It should be noted that, in this specification, an oil ring refers to a ring-shaped oil layer formed on the liquid surface within the container due to the separation of coffee oil contained in stored coffee caused by hydrophobic interactions.

[0021] Enzyme preparations can be any preparation containing lipases from the Candida genus, and can be preparations composed of lipases from the Candida genus. Furthermore, enzyme preparations can be in any form, including powder, solid, gel, and liquid.

[0022] In addition, enzyme preparations may contain a carrier, in which case the enzyme within the enzyme preparation can be immobilized on a carrier such as a porous body. In immobilized enzymes, the enzyme and carrier can be covalently bonded, attracted by electrostatic interactions, or the enzyme can be embedded within the protein in a porous body. The carrier used for immobilized enzymes is not particularly limited, but is preferably insoluble. The insoluble carrier can be inorganic or organic.

[0023] <Lipase>

[0024] Lipases are enzymes that act on triglycerides and, depending on the specificity of the enzyme, catalyze the release of fatty acids to produce diacylglycerols (DAG) or monoacylglycerols (MAG), or the reverse reaction thereof.

[0025] Examples of lipases include those from the genus *Penicillium*, *Aspergillus*, *Rhizopus*, *Candida*, *Mucor*, *Rhizopus*, and *Penicillium*. In this embodiment, the lipase is preferably from the genus *Candida*, and particularly preferably from *Candida columnarum*. In this embodiment, by using a lipase from the genus *Candida*, the formation of oil rings can be more effectively suppressed. Furthermore, in this embodiment, by using a lipase from the genus *Candida*, the formation of off-flavors (e.g., spoiled coffee odor) can also be suppressed.

[0026] Lipases can be prepared using the culture medium of microorganisms that serve as the source of the aforementioned lipases. Specific preparation methods include recovering lipases from the culture medium or bacterial cells of these microorganisms. For example, when using lipase-secreting microorganisms, the bacterial cells can be recovered from the culture medium beforehand by filtration, centrifugation, etc., and the enzyme can then be separated and / or purified. Alternatively, when using non-lipase-secreting microorganisms, the bacterial cells can be recovered from the culture medium beforehand, and the enzyme can be extracted by pulverizing the cells using pressure treatment, ultrasonic treatment, etc., and then separated and / or purified. There are no particular limitations on the method for enzyme separation and / or purification; publicly available protein separation and / or purification methods can be used, such as centrifugation, UF concentration, salting out, and various chromatographic methods using ion exchange resins. The separated and / or purified enzymes can be powdered using methods such as freeze-drying or vacuum drying. Furthermore, appropriate excipients and / or drying aids can be used in these drying methods to achieve powdering. Alternatively, appropriate additives can be added to the isolated and / or purified enzymes and then filtered and sterilized to liquefy them.

[0027] Commercially available lipases can also be used. As a preferred example of a commercially available lipase, one could mention the lipase derived from *Candida columnare* manufactured by Amano Enzyme Co., Ltd.

[0028] When the enzyme preparation is applied to coffee, the amount of lipase added relative to the total mass of the coffee is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, and even more preferably 0.05% by mass or more. Furthermore, the amount of lipase added relative to the total mass of the coffee is preferably 20% by mass or less, more preferably 10% by mass or less.

[0029] Furthermore, the amount of lipase added is not particularly limited, but the enzyme activity added relative to 1g of coffee is preferably 0.1U or more, more preferably 0.5U or more, further preferably 1U or more, even more preferably 10U or more, even more preferably 50U or more, and even more preferably 100U or more. Additionally, the lipase added relative to 1g of coffee is preferably 50,000U or less, more preferably 30,000U or less, even more preferably 10,000U or less, even more preferably 5,000U or less, even more preferably 2,000U or less, and even more preferably 1,000U or less.

[0030] The enzyme activity of lipase was determined by the following method. First, 18g of polyvinyl alcohol I and 2g of polyvinyl alcohol II were weighed and added to 800mL of water. The mixture was heated at 75-80°C for approximately 1 hour while stirring until dissolved. A 1000mL solution was prepared with water as the dissolving agent. 150mL of the emulsion and 50mL of olive oil (Japanese Pharmacopoeia) were then emulsified using a homogenizer at temperatures below 10°C to prepare an olive oil emulsion. After preparation, the olive oil emulsion was refrigerated and allowed to stand for at least 1 hour before use.

[0031] Next, mix 4 mL of McIlvaine buffer (pH 7.0) and 5 mL of olive oil emulsion, and let stand at 37°C for 10–15 minutes. Add 1 mL of enzyme solution diluted to the specified concentration, mix, and let stand at 37°C for 30 minutes. Add 10 mL of ethanol / acetone mixture (1:1), mix, and then add 10 mL of 0.05 mol / L sodium hydroxide and 10 mL of ethanol / acetone mixture (1:1), and further add phenolphthalein reagent as an indicator. While performing nitrogen replacement, stir with a stirrer and titrate with 0.05 mol / L hydrochloric acid until the pH reaches 10.00, and calculate T. 30 mL. As a blank test, 4 mL of buffer solution and 5 mL of olive oil emulsion were mixed with 10 mL of ethanol / acetone mixture (1:1), and then 1 mL of water was added. The same operation was performed to determine T0 mL. For lipase activity, the amount of enzyme that caused an increase of 1 micromole of fatty acid within 1 minute when treated at 37°C with olive oil (Japanese Pharmacopoeia) as substrate was defined as 1 unit (1 U), and was calculated using the following formula.

[0032]

[0033] <Coffee>

[0034] Coffee is in liquid or semi-liquid form. For example, coffee can be categorized as an extract obtained from coffee raw materials (green coffee beans, roasted coffee beans, or substances obtained by grinding these coffee beans) using water (including warm water and steam), or a concentrate obtained by concentrating the extract. It should be noted that a concentrate can be a semi-liquid substance such as a paste obtained by concentrating coffee. Additionally, coffee can be instant coffee, made by dissolving a dried product (such as instant coffee powder) obtained by drying the aforementioned extract or concentrate in water (including warm water and steam). Furthermore, coffee also includes a slurry formed from dispersed coffee raw materials or a solution containing coffee raw material sediment.

[0035] In this embodiment, the coffee is preferably green coffee beans, roasted coffee beans, or an extract of substances obtained by grinding these coffee beans. The type of coffee beans used is not particularly limited; examples of cultivated tree varieties include Arabica, Robusta, and Liberica, while examples of coffee varieties include Mocha, Brazilian, Colombian, Guatemalan, Blue Mountain, Kona, Mandheling, and Kilimanjaro. The coffee beans can be of a single type or a blend. In this embodiment, roasted coffee beans are preferred, and the coffee is preferably an extract of roasted coffee beans.

[0036] In this embodiment, the coffee is preferably an extract obtained from roasted coffee beans or their powdered form using water (including warm water and steam). The coffee is preferably an water (including warm water and steam) extract, and the moisture content of the coffee is preferably 80% by mass or more, more preferably 85% by mass or more, further preferably 90% by mass or more, even more preferably 95% by mass or more, and particularly preferably 99% by mass or more.

[0037] The pH of the coffee is, for example, pH 3.0 to 7.0, preferably pH 3.5 to 6.5, more preferably pH 4.0 to 6.0, and even more preferably pH 4.2 to 5.0.

[0038] In this embodiment, the coffee is preferably coffee beans or an extract of a substance obtained by crushing coffee beans. The oil content of the coffee beans is preferably 1.0% by mass or more, more preferably 1.2% by mass or more, even more preferably 1.4% by mass or more, and particularly preferably 1.5% by mass or more.

[0039] (Processed coffee)

[0040] This embodiment relates to processed coffee obtained by applying the above-described enzyme preparation to coffee. The processed coffee contains an enzyme-treated product comprising some components of coffee; in this embodiment, fatty acids are preferably included.

[0041] In this embodiment, the coffee used to activate the enzyme is preferably an aqueous extract (including warm water and steam). Therefore, the moisture content of the processed coffee is preferably 80% by mass or more, more preferably 85% by mass or more, even more preferably 90% by mass or more, and even more preferably 95% by mass or more, and particularly preferably 99% by mass or more.

[0042] The acid value of processed coffee is preferably 1.10 mg·KOH or higher. Furthermore, the acid value of processed coffee is preferably 10.0 mg·KOH or lower, more preferably 8.00 mg·KOH or lower, and even more preferably 5.00 mg·KOH or lower. The acid value of processed coffee can be calculated by neutralization titration using potassium hydroxide and a pH indicator.

[0043] It should be noted that processed coffee in this specification includes a slurry made from dispersed coffee raw materials or a solution containing coffee raw material sediment, as well as solid coffee obtained by drying processed coffee.

[0044] (Beverages containing processed coffee)

[0045] This embodiment relates to a beverage containing processed coffee, which includes the processed coffee described above. Any ingredients may be appropriately present in the beverage containing processed coffee of this embodiment. For example, as arbitrary ingredients, milk (animal milk, plant milk), flavorings, sweeteners, acidulants, bittering agents, colorings, antioxidants, pH adjusters, vitamins, amino acids, minerals, defoamers, emulsifiers (glycerol fatty acid esters, sucrose fatty acid esters, lecithin, saponins, etc.), thickening polysaccharides (pectin, carboxymethyl cellulose, etc.), salts (table salt, calcium salts, phosphates, etc.) may be added.

[0046] To further enhance the effect of inhibiting the formation of oil rings, pH adjusters can be used in beverages containing processed coffee. There are no limitations on the pH adjuster, as long as it is a substance that exhibits alkalinity when dissolved in water. Specifically, examples include baking soda (sodium bicarbonate), sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, trisodium phosphate, and tripotassium phosphate. These pH adjusters can be used individually or in combination. From the viewpoint of further enhancing the effect, baking soda (sodium bicarbonate) is preferred among these pH adjusters.

[0047] When adding a pH adjuster to a beverage containing processed coffee, there is no particular limitation on the amount of pH adjuster used. For example, the amount of pH adjuster per 100 parts by weight of coffee is preferably 1 to 1000 parts by weight, more preferably 1 to 500 parts by weight, even more preferably 1 to 100 parts by weight, and particularly preferably 1 to 50 parts by weight.

[0048] (The method of making coffee)

[0049] This embodiment relates to a method of manufacturing coffee processing, including the application of lipase from the Candida genus to coffee.

[0050] The reaction time, temperature, and pH of the reaction solution for the enzyme to act on the coffee are not particularly limited. The reaction temperature is, for example, 5–75°C, preferably 10–75°C, more preferably 15–75°C, further preferably 20–60°C, even more preferably 25–55°C, even more preferably 30–55°C, and particularly preferably 40–55°C. The pH of the reaction solution is, for example, 3.0–10.0, preferably 3.0–7.0, more preferably 4.0–7.0. The reaction time is, for example, 1 minute to 24 hours, preferably 2 minutes to 12 hours, more preferably 10 minutes to 6 hours. Using the above reaction conditions can more effectively suppress the formation of oil rings and also more effectively suppress the formation of off-flavors (e.g., spoiled coffee odor) in coffee. Furthermore, in this embodiment, when enzyme treatment is performed at a lower temperature below 15°C, the formation of oil rings can also be more effectively suppressed, and the formation of off-flavors (e.g., spoiled coffee odor) in coffee can also be more effectively suppressed. It should be noted that these reaction conditions can be appropriately selected depending on the type of coffee used, etc. It should also be noted that the optimal reaction conditions can be determined through preliminary experiments.

[0051] One embodiment of the coffee processing method includes the following steps (1) and (2). It should be noted that an enzyme inactivation step may be added after step (2).

[0052] (1) The process of preparing coffee

[0053] (2) The process of treating the prepared coffee with an enzyme preparation containing lipase from Candida spp.

[0054] By using the manufacturing method described above, it is possible to produce processed coffee that suppresses the formation of oil rings and suppresses off-flavors (e.g., spoiled coffee odor).

[0055] The coffee prepared in step (1) above is preferably an extract obtained by using water (including warm water and steam) from roasted coffee beans or their powdered form. In addition, the moisture content of the coffee prepared in step (1) is preferably 80% by mass or more, more preferably 85% by mass or more, even more preferably 90% by mass or more, even more preferably 95% by mass or more, and particularly preferably 99% by mass or more.

[0056] (Methods to inhibit aggregation)

[0057] This embodiment relates to a method for inhibiting the aggregation of coffee oil, comprising acting a lipase from the Candida genus on coffee. In this aggregation inhibition method, by acting the aforementioned enzyme preparation on coffee, the aggregation of coffee oil can be inhibited, thereby inhibiting the formation of oil rings.

[0058] Example

[0059] The following examples and comparative examples illustrate the features of the present invention in more detail. The materials, amounts, proportions, processing contents, processing steps, etc., shown in the following examples can be appropriately modified without departing from the spirit of the present invention. Therefore, the scope of the present invention should not be construed as limited to the specific examples shown below.

[0060] (Materials used)

[0061] [Table 1]

[0062]

[0063] [Table 2]

[0064]

[0065] (Enzyme activity assay method)

[0066] Weigh 18g of polyvinyl alcohol I and 2g of polyvinyl alcohol II, add 800mL of water, and heat at 75-80℃ for about 1 hour while stirring until dissolved. Prepare a 1000mL solution with water as the dissolving agent. Emulsify 150mL of the emulsion and 50mL of olive oil (Japanese Pharmacopoeia) using a homogenizer at temperatures below 10℃ to prepare an olive oil emulsion. After preparation, refrigerate the olive oil emulsion and let it stand for at least 1 hour before use.

[0067] Mix 4 mL of McIlvaine buffer (pH 7.0) and 5 mL of olive oil emulsion, and let stand at 37°C for 10–15 minutes. Add 1 mL of enzyme solution diluted to the specified concentration, mix, and let stand at 37°C for 30 minutes. Add 10 mL of ethanol / acetone mixture (1:1) and mix. Add 10 mL of 0.05 mol / L sodium hydroxide and 10 mL of ethanol / acetone mixture (1:1), and further add phenolphthalein reagent as an indicator. While performing nitrogen replacement, stir with a stirrer and titrate with 0.05 mol / L hydrochloric acid until the pH reaches 10.00. Set this titration amount as T. 30mL. As a blank test, 4 mL of buffer solution and 5 mL of olive oil emulsion were mixed with 10 mL of ethanol / acetone mixture (1:1), then 1 mL of water was added, and 0.05 mol / L hydrochloric acid was added dropwise until the pH reached 10.00. This titration volume was set as T0 mL. For lipase activity, the amount of enzyme that caused an increase of 1 micromolar fatty acid within 1 minute when treated at 37°C with olive oil (Japanese Pharmacopoeia) as a substrate was defined as 1 unit (1 U), and was calculated using the following formula.

[0068]

[0069] In the above formula, f is the coefficient of 0.05 mol / L hydrochloric acid (for quantitative measurement), and n is the dilution ratio of 1 ml of sample.

[0070] [Example 1]

[0071] (The method of making coffee)

[0072] Connect the coffee maker to a power source and press the heating button to preheat. Place 9.0g of ground coffee beans into the portafilter, level it, and then load it into the coffee maker. Once the preheated water temperature reaches 100-104°C, press the extraction button to begin extraction. Start timing from the moment the first drop of coffee (extract) flows out, and press the extraction button again at the 11-second mark to stop water flow. After extraction, coffee will continue to flow out; stop coffee recovery after 1 minute. The recovered coffee is 48.6g. Add 26.1g of hot water to adjust the Brix to 1.0%. This yields 74.7g of coffee. Repeat the above steps to obtain the desired amount of coffee.

[0073] Coffee was dispensed into 95 mL reaction vessels. Lipase from *Candida columnare* was added to the coffee at a concentration of 0.1% w / v (503 U lipase activity relative to 1 g of coffee). The reaction was carried out in a 30°C water bath for 50 minutes. After the reaction, the mixture was treated in a boiling water bath for 10 minutes to inactivate the enzyme, and a sample was collected for acid value determination. The processed coffee was then transferred to 100 mL glass bottles and stored at 5°C, observing for the formation of oil rings (aggregates of coffee oil).

[0074] [Comparative Example 1]

[0075] Coffee was prepared in the same manner as in Example 1, except that no lipase from Candida columnare was added, and coffee was obtained in the same manner as in Example 1 (control).

[0076] [Comparative Examples 2-3]

[0077] Lipase from Rhizopus deltae (Comparative Example 2) and lipase from Rhizopus oryzae (1) (Comparative Example 3) were added instead of lipase from Candida columnare. Otherwise, the processed coffee was obtained in the same manner as in Example 1.

[0078] [Evaluation 1] Oil ring formation

[0079] The coffee or processed coffee obtained in the examples and comparative examples were stored at 5°C, and the presence or absence of oil rings was observed on days 1, 7, 14, and 21. The results were evaluated according to "+++: thick oil ring formed, ++: oil ring formed, +: few oil rings formed, -: no oil ring formed".

[0080] [Table 3]

[0081] Observation of the oil ring in processed coffee

[0082]

[0083] "Evaluation 2" Calculation of relative acid value and sensory test

[0084] The acid values ​​of the coffee or processed coffee obtained in the Examples and Comparative Examples were calculated using neutralization titration with potassium hydroxide and a pH indicator. The acid value was calculated based on the weight (mg) of potassium hydroxide required to neutralize the free fatty acids contained in 1g of the sample. Furthermore, the acid value of the untreated coffee was set as 100%, and the relative acid value of the processed coffee liquid was calculated and recorded in Table 4. In addition, after storing the processed coffee at 5°C for 14 days, an evaluation of the coffee's off-odors was conducted by a panel of four members. A score was given for "strongly perceived off-odor of spoiled coffee (stale coffee smell)" (3 points), "perceived off-odor of spoiled coffee" (2 points), "slightly perceived off-odor of spoiled coffee" (1 point), and "no perceived off-odor of spoiled coffee" (0 points). The average score is recorded in Table 4 below.

[0085] [Table 4]

[0086] Sensory test results of the relative acidity of freshly enzyme-treated coffee and processed coffee stored at 5°C for 2 weeks.

[0087]

[0088] Compared to Comparative Examples 2-3, although the relative acid value increased (due to the formation of fatty acids), no off-odor was detected in Example 3. Compared to Comparative Example 2, although the relative acid value did not increase significantly, an off-odor was detected in Comparative Example 3. This is believed to be due to the substrate specificity of the enzyme causing changes in the generated fatty acids, thus affecting the perception of the off-odor.

[0089] [Example 2]

[0090] Coffee was dispensed into 95 mL reaction vessels. Lipase from *Candida columnare* was added to the coffee at a concentration of 0.1% w / v (503 U lipase activity relative to 1 g of coffee). The reaction was carried out in a 30°C water bath for 50 minutes. After the reaction, the coffee was treated in a boiling water bath for 10 minutes to inactivate the enzyme, and a sample was collected for acid value determination. The processed coffee was transferred to 100 mL glass bottles and stored at 5°C, observing for the formation of oil rings (aggregates of coffee oil). After one week of storage, the processed coffee was shaken and then stored at 5°C for further observation of oil rings.

[0091] [Comparative Example 4]

[0092] Coffee was prepared in the same manner as in Example 1, except that no lipase from Candida columnarum was added, and the coffee was observed in the same manner as in Example 2 (control).

[0093] [Evaluation 1] Oil ring formation

[0094] The coffee or processed coffee obtained in Example 2 and Comparative Example 4 were stored at 5°C, and the presence or absence of oil rings was observed on days 0, 7, 14, 21, 28, 35, 42, and 72. The results were evaluated according to the following criteria: "+++: thick oil ring formed, ++: oil ring formed, +: few oil rings formed, -: no oil ring formed".

[0095] [Table 5]

[0096] Observation of the oil ring in processed coffee

[0097]

[0098] As shown in the table above, it demonstrates that by incorporating an industrially feasible process (shaking during handling), the period during which no oil rings form can be stored for 21 days to over 28 days. It also shows that longer storage periods (72 days) are possible.

[0099] [Example 3]

[0100] The coffee solution listed in Table 1 was dispensed into 95 mL containers. Lipase from *Candida columnare* was added at a concentration of 0.05% w / v (252 U lipase activity relative to 1 g of coffee). The reaction was carried out precisely in a constant temperature bath at 10°C for 1 hour. After the reaction, the mixture was treated in a boiling water bath for 10 minutes to inactivate the enzyme. The processed coffee was then transferred to 100 mL glass bottles and stored at 5°C, observing for the formation of oil rings (coffee oil aggregates).

[0101] [Comparative Example 5]

[0102] Using the coffee liquid described in Table 1, the coffee was observed in the same manner as in Example 3 (control), except that no lipase from Candida columnarum was added.

[0103] [Evaluation 1] Oil ring formation

[0104] The coffee or processed coffee obtained in Example 3 and Comparative Example 5 were stored at 5°C, and the presence or absence of oil rings was observed on day 28. The results were evaluated according to the following criteria: "+++: thick oil ring formed, ++: oil ring formed, +: few oil rings formed, -: no oil ring formed".

[0105] [Table 6]

[0106] Observation of the oil ring in processed coffee

[0107]

[0108] It was found that lipase treatment at 10℃ also had a certain effect on reducing the formation of oil rings.

Claims

1. An enzyme preparation for inhibiting the aggregation of coffee oil, comprising a lipase from the genus Candida.

2. A method of processing coffee by applying the enzyme preparation of claim 1 to coffee.

3. The processed coffee according to claim 2, wherein, The coffee has a moisture content of 80% by mass or more.

4. A method for processing coffee, comprising acting a lipase from the genus Candida on the coffee.

5. The method of producing processed coffee according to claim 4, wherein, The coffee has a moisture content of 80% by mass or more.

6. A method for inhibiting the aggregation of coffee oil, comprising acting a lipase from the genus Candida on the coffee.

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