[논문]Functional Characteristics of Kombucha Fermented with Lactic Acid Bacteria, Yeast, and Acetic Acid Bacteria Derived from Korea Traditional Foods

Lee, Su-Min; Lee, Jae-Yong; Yoo, Dong-Gyu; Jeon, Yu-Bin; Yoon, Ho-Sik; Kim, Cheol-Hyun

doi:10.22424/jdsb.2022.40.1.23

Functional Characteristics of Kombucha Fermented with Lactic Acid Bacteria, Yeast, and Acetic Acid Bacteria Derived from Korea Traditional Foods 원문보기

Journal of dairy science and biotechnology, v.40 no.1, 2022년, pp.23 - 34

Lee, Su-Min (Department of Animal Resource & Science, Dankook University) , Lee, Jae-Yong (Department of Animal Resource & Science, Dankook University) , Yoo, Dong-Gyu (Department of Animal Resource & Science, Dankook University) , Jeon, Yu-Bin (Department of Animal Resource & Science, Dankook University) , Yoon, Ho-Sik (WellRun B&F Co.) , Kim, Cheol-Hyun (Department of Animal Resource & Science, Dankook University)

Abstract ▼ AI-Helper

In this study, to determine the importance of lactic acid bacteria (LAB) in Kombucha fermentation, biological functions, such as organic acid production and anti-inflammatory and antibacterial activities, of Kombucha, with or without LAB inoculation, were evaluated. Lactobacillus paracasei DK215, Saccharomyces cerevisiae C3, and Acetobacter pasteurianus P2 were selected as the inoculants. Organic acids were measured every 3 days from the end of fermentation using HPLC; the organic acid content of LAB-inoculated Kombucha was relatively high. Samples with or without LAB inoculation showed high antibacterial activity against Escherichia coli. The MTT assay results indicated no significant difference in concentration difference and cell death. In the NO production test, compared with the uninoculated Kombucha sample, the LAB-inoculated Kombucha sample exhibited a value similar to that of the group without LPS treatment. The levels of cytokine (IL-1α, IL-6, TNF-α) production were significantly lower than those of the LPS(+) group, indicating the anti-inflammatory activity potential of the Kombucha sample. This improvement in the biological function of the LAB-inoculated Kombucha further verifies the value of LAB in the fermented food and beverage industry.

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표/그림 (10)

표 Table 1. Instrument and operating conditions of HPLC
그림 Fig. 1. Determination of organic acid standards using HPLC. Absorbance at 210 nm.
그림 Fig. 2. Separation of organic acid content on 1st, 4th, 7th day of fermentation of KWOL.
표 Table 2. Organic acid standard
그림 Fig. 3. Separation of organic acid content on 1st, 4th, 7th day of fermentation of KWL.
표 Table 3. Antibacterial activities of KWOL
그림 Fig. 4. Effect of Kombucha on cell viability in RAW 264.7 cells. All values were mean±SD of triplicates. 100, 10, 1, 0.1, 0.01, 0.001 mg/mL: Acetic acid content of the sample. * p<0.05, ** p< 0.01, *** p<0.001.
표 Table 4. Antibacterial activities of KWL
그림 Fig. 5. Effect of Kombucha on nitric oxide in LPS activated RAW 264.7 cells. All values were mean±SD of triplicates. 100, 10, 1, 0.1, 0.01, 0.001 mg/mL: Acetic acid content of the sample. * p<0.05, ** p<0.01, *** p<0.001. LPS, lipopolysaccharide.
그림 Fig. 6. Comparison of Cytokines (IL-1α, IL-6, TNF-α) production on Kombucha in LPS-unstimulated RAW264.7 cells. All values were mean±SD of triplicates. 0.1 mg/mL: Acetic acid content of the sample. * p<0.05, ** p<0.01, *** p<0.001. LPS, lipopolysaccharide.

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제안 방법

After mixing the strain (Lactobacillus paracasei DK215, Saccharomyces cerevisiae C3, and Acetobacter pasteurianus P2) diluted to 1×10³ CFU/mL in the sterilized solution, the inlet was sealed using sterile gauze, and aerobic fermentation and stationary fermentation were performed at 30℃.
Therefore, in this study, the importance of LAB in Kombucha fermentation was evaluated, and biological functions such as organic acid production, anti-inflammatory, antibacterial activity, and antioxidant activity of kelp were evaluated according to whether or not LAB were inoculated. Also, instead of using green tea and black tea, which are commonly used as raw materials for Kombucha, the experiment was conducted using green tea extract as a substrate. Therefore, Kombucha that was not inoculated with LAB was named Kombucha without Lactic acid bacteria (KWOL), and Kombucha that was inoculated with LAB was named Kombucha with Lactic acid bacteria (KWL).
In addition, the vitamin B complex secreted by AAB provides a favorable environment for the growth of other yeasts and LAB in kefir grains [20]. Therefore, in this study, the importance of LAB in Kombucha fermentation was evaluated, and biological functions such as organic acid production, anti-inflammatory, antibacterial activity, and antioxidant activity of kelp were evaluated according to whether or not LAB were inoculated. Also, instead of using green tea and black tea, which are commonly used as raw materials for Kombucha, the experiment was conducted using green tea extract as a substrate.
This study was conducted to evaluate the potentially beneficial biological functions of Kombucha inoculated with LAB, such as organic acid production, anti-inflammatory, antibacterial activity and antioxidant capacity, compared to Kombucha normally fermented using yeast and AAB. Lactobacillus paracasei DK215, Saccharomyces cerevisiae C2, Acetobacter pasteurianus P2 was selected and used in the experiment.

대상 데이터

This study was conducted to evaluate the potentially beneficial biological functions of Kombucha inoculated with LAB, such as organic acid production, anti-inflammatory, antibacterial activity and antioxidant capacity, compared to Kombucha normally fermented using yeast and AAB. Lactobacillus paracasei DK215, Saccharomyces cerevisiae C2, Acetobacter pasteurianus P2 was selected and used in the experiment.

이론/모형

In this study, the antibacterial activity of Kombucha against various pathogens was investigated using the agar diffusion method
The antimicrobial activity of Kombucha tea samples was assessed by agar diffusion assay [21] against four human pathogenic bacteria Gram-positive Staphylococcus aureus (KCTC3881, Korea) and Gram-positive bacilli Bacillus cereus (KCTC362A) and Gram-negative bacilli including E. coli (KCTC1682), Salmonella enterica (KCTC2054).

성능/효과

After culturing RAW264.7 cells, which are mouse macrophages, in 96 wells at a level of 1×105 cells per well, the cytotoxicity MTT assay results of samples diluted by acetic acid concentration showed cytotoxicity or growth at the remaining concentrations except for 100 mg/mL
enterica were added and compared with various control groups. As a result of the experiment, it was confirmed that it exhibits antibacterial activity in S. aureus, E.
Cytokine (IL-1α, IL-6, TNF-α) production was significantly lower than that of the LPS (+) group, and based on this result, it was possible to find the possibility of anti-inflammatory activity in the Kombucha sample.
LPS is a lipopolysaccharide, a structure that exists on the surface of bacteria, and is a substance that can be used as an antigen to induce an immune response in the host. NO is a substance produced by such an immune response, and when looking at the amount of production, it was confirmed that the Kombucha fermented sample had a lower NO production than the LPS-treated group. In addition, compared with KWOL, it was confirmed that the value of the KWL sample was close to that of the LPS untreated group.
Glucuronic acid is the most important metabolite during Kombucha fermentation, improving immune response, anticancer, gastric ulcer treatment and antidiabetic are effective [26–29]. Therefore, in this experiment, as a result of measuring the content of a total of five organic acids including glucuronic acid at 3-day intervals during Kombucha fermentation, it was confirmed that 7.6 g/L of acetic acid on the 7th day of fermentation was reached in KWL, and Glucuronic acid was also It was confirmed that it reached 7 g/L. In the case of KWOL, the organic acid content was relatively low at 3.
Therefore, in this study, it was judged that inoculation of Lactobacillus paracaseiDK215 improved the antibacterial and the change of glucuronic acid content during fermentation, compared to the original Kombucha using only yeast and AAB
cereus. Unfermented tea had no antibacterial activity against target microorganisms, but both Kombucha samples showed the highest level of antimicrobial activity in E. coli, and relatively higher antimicrobial activity was confirmed in KWL.

후속연구

It has been confirmed that the improvement of the biological function of Kombucha inoculated with these LAB can once again prove the value of the LAB in the fermented food and beverage industry. However, further investigation into the mechanisms involved in the fermented beverage raw material substrates involved in the production of biological metabolites is deemed necessary.
Therefore, in this study, it was judged that inoculation of Lactobacillus paracaseiDK215 improved the antibacterial and the change of glucuronic acid content during fermentation, compared to the original Kombucha using only yeast and AAB. It has been confirmed that the improvement of the biological function of Kombucha inoculated with these LAB can once again prove the value of the LAB in the fermented food and beverage industry. However, further investigation into the mechanisms involved in the fermented beverage raw material substrates involved in the production of biological metabolites is deemed necessary.

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