KR101738303B1 - Method for producing coffee green beans with enhanced chlorogenic acid content by germination and fermentation - Google Patents

Abstract

The present invention relates to a method for producing coffee green beans with enhanced chlorogenic acid content, coffee green beans with enhanced chlorogenic acid content manufactured by the method and a method for improving chlorogenic acid content in coffee green beans. The coffee green beans with enhanced chlorogenic acid content is manufactured by the following steps: (a) disinfecting Arabica coffee green beans, soaking the same in water and germinating the same; (b) sterilizing the germinated coffee green beans in the step (a); (c) inoculating Bacillus subtilis strains to the sterilized coffee green beans in the step (b) and fermenting the same; and (d) drying the fermented coffee green beans in the step (c).

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for producing coffee beans by germination and fermentation with improved chlorogenic acid content,

(A) disinfecting arabica seed coffee bean and then germinating by soaking in water; (b) sterilizing the germinated coffee bean germinated in step (a); (c) adding the sterilized coffee bean of step (b) to Bacillus subtilis subtilis ) and then fermenting the same; And (d) drying the fermented coffee beans of step (c). The method of producing coffee beans having improved chlorogenic acid content, the method of producing coffee beans having improved chlorogenic acid content prepared by the method, And a method for enhancing the chlorogenic acid content in coffee beans.

Coffee Rubiaceae (Rubiaceae) belonging to the subgenus kope (Coffee), varieties are grown commercially significant arabica (Arabica) and Robusta-divided into (Robusta canephora), Riverina and South (Liberica) 3 varieties. Coffee is a typical drink that is made by harmony of bitter taste, bitter taste, sour taste, and sweetness. It is the most widely consumed drink in the world. In Korea, the coffee market is continuously growing, such as proliferation of coffee specialty stores and increase in consumer consumption.

Coffee is well known to reduce the risk of neurological diseases such as Alzheimer's and Parkinson's and other metabolic diseases caused by oxidative stress as well as mental effects such as arousal and mood swings. Proper coffee intake has many positive effects on the health of adults, except children and pregnant women. It contains various physiologically active ingredients such as polyphenol compounds and tocopherol, and it contains chlorogenic acid, caffeine ), Melanoidin, and the like. In particular, chlorogenic acid is based on antioxidant activity and has functionalities such as prevention of liver fibrosis which is a leading index of cirrhosis. Therefore, there is a growing interest in health and consumers' demand for coffee containing natural antioxidants is on the rise.

Germination is sprouted on seeds. It is not easy to chew and hard to digest when eating grain, but when germinated, it softens and various digestive enzymes are produced, resulting in higher nutrient uptake rate. In addition, soluble fiber is three times as much as ordinary grains. Various vitamins, amino acids , Enzymes and other physiologically active substances are increased, so they are helpful in nutrition, and the content of antioxidants increases during germination.

Fermented foods are foods made by utilizing the fermentation action of microorganisms such as lactic acid bacteria and yeast. Depending on the kind of microorganism and the material of the food, the fermented food is various and has a unique characteristic and flavor depending on the kind of the fermented food. The functionality of the food raw material can be improved.

Korean Patent Laid-Open Publication No. 2016-0025090 discloses a method for producing fermented coffee. Korean Patent Registration No. 1014871 discloses a method for producing a fermented fermented coffee fermented with Kimchi lactic acid bacteria. However, the chlorinated acid content of the present invention This method is different from the method of producing coffee beans by germination and fermentation.

The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a coffee bean product which is optimized for manufacturing conditions such as germination, fermentation, sterilization and drying conditions so as to improve the content and flavor of chlorogenic acid while reducing the bitter taste, Which is excellent in flavor and palatability without increasing fermentation odor, and has improved chlorogenic acid content.

In order to solve the above-described problems, the present invention provides a method for producing a safflower seedlings comprising: (a) disinfecting arabica seed coffee bean and then germinating by soaking in water; (b) sterilizing the germinated coffee bean germinated in step (a); (c) adding the sterilized coffee bean of step (b) to Bacillus subtilis subtilis ) and then fermenting the same; And (d) drying the fermented coffee bean of the step (c). The present invention also provides a method for producing coffee beans having improved chlorogenic acid content.

Further, the present invention provides a coffee bean having improved chlorogenic acid content prepared by the above method.

The present invention also relates to a method for producing ginseng, comprising the steps of: (a) sterilizing arabica seed coffee bean and then germinating by soaking in water; (b) sterilizing the germinated coffee bean germinated in step (a); (c) adding the sterilized coffee bean of step (b) to Bacillus subtilis subtilis ) and then fermenting the same; And (d) drying the fermented coffee beans of step (c) to increase the content of chlorogenic acid. The present invention also provides a method for enhancing the content of chlorogenic acid in coffee beans.

The coffee green bean prepared by optimizing the fermentation strain of the present invention, the germination condition, the sterilization condition, the fermentation condition and the like has a high content of chlorogenic acid and is excellent in taste, flavor and preference without fermentation, Coffee beans can be provided.

Figure 1 is a schematic representation of the process of germination and germination of coffee bean.
Fig. 2 shows the 10 day germination of coffee bean.

In order to achieve the object of the present invention,

(a) disinfecting arabica seed coffee bean and then germinating by soaking in water;

(b) sterilizing the germinated coffee bean germinated in step (a);

(c) inoculating and boiling the Bacillus subtilis strain in the sterilized coffee bean of step (b); And

and (d) drying the fermented coffee bean of the step (c). The present invention also provides a method of producing coffee beans having improved chlorogenic acid content.

In the method of manufacturing coffee beans according to the present invention, the disinfection of step (a) can be preferably performed by immersing the coffee bean in 0.8 to 1.2% (v / v) sodium hypochlorite solution for 1 to 3 minutes, More preferably, coffee beans can be sterilized by immersing them in a 1% (v / v) sodium hypochlorite solution for 2 minutes. Disinfecting coffee beans with the above-mentioned conditions can effectively sterilize the coffee beans while maintaining the quality of the coffee beans. However, when the concentration and immersion time of the solution are less than the above range, there is a disadvantage that the effect of sterilizing is insignificant. If the time exceeds the above range, there is a problem that nutritional components of coffee bean are eluted.

Further, in the method for producing coffee beans of the present invention, the germination in step (a) may preferably be germinated at 28 to 32 ° C for 12 to 18 days, more preferably at 30 ° C for 15 days . It was possible to further increase the chlorogenic acid content of the prepared coffee bean sprouts compared to germinating the coffee bean sprouts under the above conditions at different temperatures and periods.

In addition, in the method for producing coffee beans of the present invention, the sterilization in step (b) can be sterilized preferably at 110 to 130 ° C for 10 to 20 minutes, more preferably at 121 ° C for 15 minutes . The germinated green beans were fermented after sterilization under the same conditions as above, so that the germinated green beans could be further enhanced in chlorogenic acid content compared to fermenting them without sterilization.

Further, in the method for producing coffee beans of the present invention, the inoculation of the step (c) is preferably carried out by a Bacillus subtilis subtilis) which coffee green beans sterilized strain of 8% to 12%, based on the weight of (v / w), preferably 10% (v / w) can be given. The chlorogenic acid content of the fermented green beans was higher than that inoculated with the strain under the above conditions, and the preference of the prepared coffee could be improved.

Further, in the method for producing coffee beans of the present invention, the fermentation in the step (c) may preferably be carried out at 38 to 42 ° C for 44 to 52 hours, more preferably at 40 ° C for 48 hours . Fermentation under the same conditions as above was reduced in fermentation yield and could be made from coffee bean which was further enhanced in chlorogenic acid content.

Further, in the process for producing coffee beans of the present invention, drying in step (d) may preferably be performed at 55 to 65 ° C for 16 to 20 hours, more preferably at 60 ° C for 18 hours . The dried coffee beans fermented under the above conditions could further improve the flavor of the prepared coffee as the content of chlorogenic acid was further increased.

The method for producing coffee beans having improved chlorogenic acid content of the present invention is more specifically

(a) dipping arabinogaster coffee bean sprouts in a 0.8-1.2% sodium hypochlorite solution for 1-3 minutes, soaking them in water at 28-32 ° C for 12-18 days;

(b) sterilizing the germinated coffee bean of step (a) at 110 to 130 ° C for 10 to 20 minutes;

(c) 8 ~ 12% ( v / w) 44 ~ 52 eseo inoculated after 38 ~ 42 ℃ was sterilized sterilized Bacillus subtilis (Bacillus subtilis) strain in which coffee green beans coffee green beans, based on the weight of the step (b) Fermenting for a period of time; And

(d) drying the fermented coffee bean of step (c) at 55 to 65 ° C for 16 to 20 hours,

More specifically,

(a) dipping arabinogam coffee beans in 1% hypochlorous acid sodium solution for 2 minutes, immersing in water, germinating at 30 ° C for 15 days;

(b) sterilizing the germinated coffee bean of step (a) at 121 DEG C for 15 minutes;

(c) inoculating 10% (v / w) of the coffee ground beef sterilized with the Bacillus subtilis strain to the sterilized coffee bean of step (b), and then fermenting at 40 ° C for 48 hours; And

(d) drying the fermented coffee bean of step (c) at 60 DEG C for 18 hours.

The present invention also provides a coffee bean having improved chlorogenic acid content prepared by the above method.

The present invention also relates to

(a) dipping arabinogaster coffee bean sprouts in a 0.8-1.2% sodium hypochlorite solution for 1-3 minutes, soaking them in water at 28-32 ° C for 12-18 days;

(b) sterilizing the germinated coffee bean of step (a) at 110 to 130 ° C for 10 to 20 minutes;

(c) 8 ~ 12% ( v / w) 44 ~ 52 eseo inoculated after 38 ~ 42 ℃ was sterilized sterilized Bacillus subtilis (Bacillus subtilis) strain in which coffee green beans coffee green beans, based on the weight of the step (b) Fermenting for a period of time; And

(d) drying the fermented coffee bean of step (c) at 55 to 65 ° C for 16 to 20 hours to increase the content of chlorogenic acid, thereby providing a method for enhancing the content of chlorogenic acid in coffee beans.

The method of enhancing the chlorogenic acid content in the coffee bean of the present invention is more specifically

(a) dipping arabinogam coffee beans in 1% hypochlorous acid sodium solution for 2 minutes, immersing in water, germinating at 30 ° C for 15 days;

(b) sterilizing the germinated coffee bean of step (a) at 121 DEG C for 15 minutes;

(c) inoculating 10% (v / w) of the coffee ground beef sterilized with the Bacillus subtilis strain to the sterilized coffee bean of step (b), and then fermenting at 40 ° C for 48 hours; And

(d) drying the fermented coffee bean of step (c) at 60 DEG C for 18 hours.

Hereinafter, embodiments of the present invention will be described in detail. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Manufacturing example  1: germination - sterilization - fermentation - coffee by drying Green bean  Produce

(a) The Arabica coffee green beans (Laos' Club Green Coffee) were stored at room temperature before use in the experiment, and the parchment was stripped manually by hand before germination experiments. The green beans from which the parchment was removed were immersed in a 1% (v / v) sodium hypochlorite solution for 2 minutes, sterilized, and rinsed with tap water filtered with a bacterial filter (0.22 μm). Test germination using a Petri dish was carried out under the conditions in an incubator, and germinated at 30 ± 1 ° C for 15 days while feeding the coffee beans twice a day with water filtered at a ratio of 1: 1 (w: v).

(b) The germinated coffee bean germinated in the step (a) was sterilized at 121 캜 for 15 minutes.

(c) The sterilized coffee bean of step (b) is cooled to 40 DEG C, and Bacillus subtilis Subtilis strain (3 × 10 ⁶ CFU / ml) was inoculated by 10% (v / w) of the weight of the sterilized coffee bean and fermented at 40 ° C. for 48 hours.

(d) The fermented coffee bean of step (c) was washed in running water and dried in a hot air drier at 60 ° C for 18 hours.

Comparative Example  1: Coffee by germination-drying Green beans  Produce

(a) The Arabica coffee green beans (Laos' Club Green Coffee) were stored at room temperature before use in the experiment, and the parchment was stripped manually by hand before germination experiments. The green beans from which the parchment was removed were immersed in a 1% (v / v) sodium hypochlorite solution for 2 minutes, sterilized, and rinsed with tap water filtered with a bacterial filter (0.22 μm). Test germination using a Petri dish was carried out under the conditions in an incubator, and germinated at 30 ± 1 ° C for 15 days while feeding the coffee beans twice a day with water filtered at a ratio of 1: 1 (w: v).

(b) The germinated coffee bean germinated in step (a) was washed in flowing water and dried in a hot air dryer at 60 ° C for 18 hours.

Comparative Example  2: Sterilization - Fermentation - Coffee by drying Green beans  Produce

(a) Water was added to arabica coffee green beans (Laos's Club Green Coffee) at a ratio of 1: 1 (w: v) to immerse the coffee beans in water completely and sterilized at 121 ° C for 15 minutes.

(b) wherein (a) and then step in the sterilization was coffee green beans were sterilized for Bacillus subtilis (Bacillus subtilis) strain coffee green beans 10% by weight (v / w) inoculation a mixture of was fermented at 40 ℃ for 48 hours. .

(c) The fermented coffee bean of step (b) was washed in flowing water and dried in a hot air dryer at 60 ° C for 18 hours.

Comparative Example  3: Coffee by germination-fermentation-drying Green bean  Produce

(a) The Arabica coffee green beans (Laos' Club Green Coffee) were stored at room temperature before use in the experiment, and the parchment was stripped manually by hand before germination experiments. The green beans from which the parchment was removed were immersed in a 1% (v / v) sodium hypochlorite solution for 2 minutes, sterilized, and rinsed with tap water filtered with a bacterial filter (0.22 μm). Test germination using a Petri dish was carried out under the conditions in an incubator, and germinated at 30 ± 1 ° C for 15 days while feeding the coffee beans twice a day with water filtered at a ratio of 1: 1 (w: v).

(b) 10% (v / w) of the germinated coffee bean germ in the step (a) was inoculated with the Bacillus subtilis strain germinated on the weight of the germinated coffee bean, and the mixture was fermented at 40 ° C for 48 hours .

(c) The fermented coffee bean of step (b) was washed in flowing water and dried in a hot air dryer at 60 ° C for 18 hours.

Experimental Method

(1) Content of chlorogenic acid

The content of chlorogenic acid was obtained by adding 1 ml of boiled water to 100 mg of ground coffee ground powder and mixing well for 1 minute. After allowing to stand for 5 minutes, centrifuge at 10,000 × G for 2 minutes. Take 100 μl of supernatant, add 5 μl of Carrez 1 solution, mix 5 μl of Carrez 2 solution, and mix. Then, 30 μl of methanol was mixed and centrifuged at 10,000 × G for 2 minutes. After centrifugation, 50 占 퐇 of the supernatant was diluted 10 占 to 450 占 퐇 Solvent A (distilled water) and 1 占 퐇 was taken and injected into a high performance liquid chromatograph (HPLC) to analyze chlorogenic acid. Qualitative and quantitative analysis was performed at 325 nm, the maximum absorption wavelength of chlorogenic acid. At this time, acetonitrile (solvent B) was used as a mobile phase. In addition, standard solution of chlorogenic acid standard solution was prepared by dissolving chlorogenic acid standard product in methanol and diluting the standard stock solution to 1,000 ppm, so as to be 100 ppm, as standard solution for calibration curve.

(2) Caffeine content

The caffeine content was analyzed by the same method as that of chlorogenic acid, but the qualitative and quantitative analysis was performed at 272 nm, the maximum absorption wavelength of caffeine. In addition, as the caffeine standard solution, a standard stock solution of caffeine standard dissolved in methanol was diluted to 1,000 ppm to obtain 100 ppm, which was used as the standard solution for the calibration curve.

Example  1: coffee Green bean Chlorogenic acid  And caffeine content

The results of the comparison of chlorogenic acid and caffeine contents of coffee bean sprouts prepared by the methods of Preparation Example 1 and Comparative Examples are shown in Table 1 below. The contents of chlorogenic acid and caffeine increased in% compared to untreated coffee beans.

Chlorogenic acid and caffeine content of coffee bean (%) division Chlorogenic acid (%) Caffeine(%) Green beans 100 100 Production Example 1 245 125 Comparative Example 1 170 130 Comparative Example 2 120 100 Comparative Example 3 182 112

As a result, there was no significant difference in the caffeine content of coffee beans from germinated and fermented treatments compared with untreated coffee beans. However, the content of chlorogenic acid in coffee beans increased significantly . In particular, it was confirmed that when the germinated coffee beans as in Comparative Example 1 and the coffee beans fermented as in Comparative Example 2 were subjected to both germination and fermentation processes, the content of chlorogenic acid was remarkably increased. In addition, it was confirmed that the coffee bean sprouts prepared through germination-sterilization-fermentation process as in Production Example 1 had a higher chlorogenic acid content than the coffee bean sprouts fermented immediately after germination as in Comparative Example 3.

Example  2: Sensory evaluation of coffee

20 people who are women (19 ~ 64 years), 20 people who are men (19 ~ 64 years), 5 women who are 65 years or older and 5 men who are 65 years or older , Flavor, body sensation, aftertaste and overall evaluation. (1 point: very poor, 2 points: poor, 3 points: 1 point: poor, 2 points: poor, and 3 points). The coffee grounds of Preparation Example 1 and Comparative Example were roasted, ground, ground, Normal, 4 points: good, 5 points: very good). The results are shown in Table 2.

Sensory evaluation of coffee division Sour taste sweetness bitter Flavor Body feel aftertaste Overall Production Example 1 4.0 3.7 3.8 4.2 4.1 4.2 4.1 Comparative Example 1 3.8 3.4 3.2 3.5 3.6 3.8 3.6 Comparative Example 2 3.7 3.5 3.1 3.4 3.3 3.5 3.5 Comparative Example 3 3.7 3.6 3.3 3.6 3.7 3.6 3.7

As a result, it was confirmed that the coffee using the coffee bean of Preparation Example 1 showed a score of 4 or higher in all the items, which is preferable to the comparative examples by the evaluators. Accordingly, it was found that the preference of coffee was improved by preparing coffee bean sprouts by the method of Production Example 1.

Example  3: Coffee according to germination condition Green bean Chlorogenic acid  content

Table 3 shows the results of comparing the chlorogenic acid content of the coffee beans produced by varying the germination temperature and the germination period in the step (a). The amount of chlorogenic acid was increased in% compared to untreated green beans.

Chlorogenic acid content of coffee bean according to germination condition (%) Germination temperature Germination period 5 days 10 days 15th 20 days 25 ℃ 161 190 206 204 30 ℃ 162 202 245 224 35 ℃ 158 200 212 220

The chlorogenic acid content of germinated coffee bean increased with germination at 25 ~ 30 ℃, but the chlorogenic acid content decreased with germination at 20th germination. According to germination temperature, the highest value of chlorogenic acid content was found in coffee bean germinated at 30 ℃ compared to other temperature conditions. As a result, it was confirmed that it is preferable to ferment after germination at 30 ° C for 15 days in order to prepare a fermented coffee bean having a greatly increased chlorogenic acid content.

Example  4: Coffee according to fermentation strain Green bean Chlorogenic acid  content

A coffee bean product was prepared by the method of Preparation Example 1, except that in step (c), Bacillus subtilis subtilis) not inoculated with the strain Bacillus Lee Kenny Po Ms (Bacillus licheniformis ) (Comparative Example 4), Lactobacillus ( Lactobacillus The results are shown in Table 4 below. The results are shown in Table 4 below. The amount of chlorogenic acid was increased in% compared to untreated green beans.

Chlorogenic acid content of coffee bean according to fermentation strain (%) division Chlorogenic acid (%) Production Example 1 245 Comparative Example 4 147 Comparative Example 5 109

As a result of comparing the chlorogenic acid content of the coffee beans according to the fermentation strain, the Bacillus subtilis strain was inoculated into the fermented coffee beans (Comparative Examples 4 and 5) by fermentation using Bacillus licheniformis or Lactobacillus flutarium, The coffee bean of one Preparation Example 1 showed a significantly higher chlorogenic acid content. As a result, it was confirmed that it is preferable to inoculate Bacillus subtilis strain to ferment the germinated and fermented coffee bean which has a greatly increased chlorogenic acid content.

Example  5: coffee according to strain inoculation Green bean Chlorogenic acid  content

Table 5 shows the results of comparing the chlorogenic acid content of coffee beans prepared by varying the inoculation amount of Bacillus subtilis strain in the step (c) of preparing coffee bean by the method of Preparation Example 1, The amount of chlorogenic acid was increased in% compared to untreated green beans.

Chlorogenic acid content (%) of coffee beans according to strain inoculum Amount of strain inoculated Content of chlorogenic acid (%) 5% 215 10% 245 15% 222

As a result of comparing the content of chlorogenic acid of coffee bean according to the inoculation amount of fermentation strain, it was confirmed that the content of chlorogenic acid was increased compared to that of 5% and 15% fermentation by adding 10% of Bacillus subtilis strain to coffee ground beef, And finally adding 10% of Bacillus subtilis strain.

Example  6: Coffee according to fermentation conditions Green bean Chlorogenic acid  content

Table 6 below shows the results of comparing the chlorogenic acid content of coffee beans having different fermentation temperature and fermentation time in the step (c) of preparing coffee bean by the method of Preparation Example 1, respectively. The amount of chlorogenic acid was increased in% compared to untreated green beans.

Chlorogenic acid content of germinated and fermented coffee bean according to fermentation conditions (%) Fermentation temperature Fermentation time 24 hours 36 hours 48 hours 60 hours 37 ℃ 161 190 206 204 40 ℃ 162 202 245 224 43 ℃ 158 200 212 220

According to the fermentation conditions, the content of chlorogenic acid in coffee bean sprouts was the lowest and that of coffee sprouted fermented at 37 ℃ showed the lowest chlorogenic acid content. The fermented coffee bean fermented at 40 ℃ showed the highest chlorogenic acid content. The content of chlorogenic acid increased until fermentation 48 hours after fermentation but decreased after 60 hours fermentation. As a result, it was confirmed that fermentation at 40 ° C for 48 hours is preferable in order to prepare coffee beans having a greatly increased content of chlorogenic acid.

Example  7: Coffee according to drying conditions Green bean Chlorogenic acid  content

Table 7 below shows the results of comparing the chlorogenic acid content of coffee beans with different drying temperature and drying time in the step (d) of preparing coffee bean by the method of Preparation Example 1. The amount of chlorogenic acid was increased in% compared to untreated green beans.

Chlorogenic acid content of germinated and fermented coffee beans according to drying conditions (%) Drying temperature Drying time 6 hours 12 hours 18 hours 24 hours 50 ℃ 181 214 222 220 60 ° C 192 212 245 228 70 ℃ 189 220 225 224

As a result of comparing the chlorogenic acid contents of coffee bean seeds according to the drying conditions, coffee seeds dried at 50 ℃ showed the lowest chlorogenic acid content, and coffee seeds dried at 60 ℃ showed the highest chlorogenic acid content. The drying time was determined by drying at 60 ° C for 18 hours in order to prepare a coffee bean having a high content of chlorogenic acid with a high chlorogenic acid content of coffee beans dried for 18 hours.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

<The present invention is performed thanks to the service cost of Goheung Niche crop agricultural product diversification project of Gohung Local Food Local Business Center>

Claims (5)

(a) dipping arabinogaster coffee bean sprouts in a 0.8-1.2% sodium hypochlorite solution for 1-3 minutes, soaking them in water at 28-32 ° C for 12-18 days;
(b) sterilizing the germinated coffee bean of step (a) at 110 to 130 ° C for 10 to 20 minutes;
(c) 8-12% (v / w) inoculated with the coffee grounds sterilized with 3 × 10 ⁶ CFU / ml of Bacillus subtilis strain in the sterilized coffee bean of step (b) Followed by fermentation for 44 to 52 hours at 38 to 42 DEG C; And
(d) drying the fermented coffee bean of step (c) at 55 to 65 ° C for 16 to 20 hours. delete delete A coffee bean having improved chlorogenic acid content prepared by the method of claim 1. (a) dipping arabinogaster coffee bean sprouts in a 0.8-1.2% sodium hypochlorite solution for 1-3 minutes, soaking them in water at 28-32 ° C for 12-18 days;
(b) sterilizing the germinated coffee bean of step (a) at 110 to 130 ° C for 10 to 20 minutes;
(c) 8-12% (v / w) inoculated with the coffee grounds sterilized with 3 × 10 ⁶ CFU / ml of Bacillus subtilis strain in the sterilized coffee bean of step (b) Followed by fermentation for 44 to 52 hours at 38 to 42 DEG C; And
(d) drying the fermented coffee bean of step (c) at 55 to 65 ° C for 16 to 20 hours to increase the content of chlorogenic acid in the coffee beans.

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