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Bioscience, Biotechnology, and Biochemistry Volume 84, 2020 - Issue 9
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Microbiology & Fermentation Technology

Changes in lactic acid bacteria and components of Awa-bancha by anaerobic fermentation

Hiroki Nishiokaa Food and Biotechnology Division, Tokushima Prefectural Industrial Technology Center, Tokushima, Japan;b United Graduate School of Agricultural Science, Gifu University, Gifu, Japan;c Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, JapanCorrespondence[email protected]
ORCID Iconhttps://orcid.org/0000-0002-0487-902X
ORCID Icon,
Tomofumi Mizunod The Graduate School of Natural Science and Technology, Gifu University, Gifu, Japan
,
Hitoshi Iwahashie Faculty of Applied Biological Sciences, Gifu University, Gifu, JapanORCID Iconhttps://orcid.org/0000-0003-1100-5977
ORCID Icon &
Masanori Horiec Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Takamatsu, JapanORCID Iconhttps://orcid.org/0000-0001-9640-9350
ORCID Icon
Pages 1921-1935 | Received 13 Mar 2020, Accepted 11 May 2020, Published online: 28 May 2020

Figures & data

Figure 1. The production area and production process of Awa-bancha.

Awa-bancha is produced in Kamikatsu and Naka in Tokushima. In the early summer, tea leaves are harvested by hand. The leaves are plucked from the root. The tea leaves are then boiled in large kettles for sterilization and enzyme deactivation. They are then kneaded by a kneading machine. The tea leaves are placed in a barrel, heavy stones are placed on them, and then the broth is added. After 2 to 4 weeks, the tea leaves are removed from the barrel and dried in the sun to complete the process.
Figure 1. The production area and production process of Awa-bancha.

Table 1. Samples collected from different producer.

Figure 2. Number of bacteria in each production process.

The wet tea leaves and sterilized PBS were mixed and suspended at a concentration of 0.1 mg/mL. The suspension was diluted with sterile PBS and spread on each agar plate.Standard agar plates were cultured at 37°C for 2 days. YM agar plates was cultured at 30°C for 5 days. MRS agar plates was cultured in an anaerobic jar at 37°C for 2 days. After the culture process, the colonies were counted.
Figure 2. Number of bacteria in each production process.

Figure 3. Analysis of the bacterial flora by 16 S rRNA genes using a next-generation sequencer.

(a) Raw tea leaves and (b) Tea leaves after anaerobic fermentation.Bacteria that accounted for less than 5% were grouped as others.
Figure 3. Analysis of the bacterial flora by 16 S rRNA genes using a next-generation sequencer.

Table 2. Number of bacteria isolated by MRS agar plates in each product process.

Figure 4. The proportion of bacteria isolated by MRS agar plates in each production process.

Bacteria that accounted for less than 5% were grouped as others.
Figure 4. The proportion of bacteria isolated by MRS agar plates in each production process.

Table 3. Growth of lactic acid bacteria isolated on green tea extract medium containing 1% glucose.

Table 4. Sugar utilization of Lactobacillus pentosus isolated.

Table 5. Contents of organic acids and pH in Awa-bancha.

Table 6. Contents of free amino acids in Awa-bancha.

Table 7. Contents of caffeine and catechins in Awa-bancha.

Figure 5. Evaluation of degranulation by the tea extract for RBL-2H3 cells.

Degranulation was induced in RBL-2H3 cells in the presence of Awa-bancha extract, and the inhibitory effect was evaluated. The value is a relative value where the release rate of cells not containing Awa-bancha extract is 100% (n = 6). **p < 0.01.
Figure 5. Evaluation of degranulation by the tea extract for RBL-2H3 cells.

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