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Classification of microorganisms and generarion of -amino butylic acid from the post-heated fermented tea

后热发酵茶中微生物的分类及β-氨基丁酸的产生

基本信息

批准号：
18500603
负责人：
KATOH Miyuki
金额：
$ 2.53万
依托单位：
Kagawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Post-heated fermented teas were sampled during their production processes to study properties of microorganisms isolated from the samples. Unlike NBRC9405^T, a fungus isolate from Ishizuchi-kurocha had no rhizoid. Sporangiospores of NBRC31005 formed lamellae. The D2 region of a 28S rRNA gene from the fungus isolate was sequenced to find 98.7% homology with that of Rhizomucor variabilis.Bacillus circulans isolates from Awa-bancha, Ishizuchi-kurocha, and Goishi-cha were examined for optimum temperature and pH. The optimum temperatures were 37-40℃ and the optimum pH were 6-7. Components of the post-heated fermented teas were found to suppress the isolates in growth. Both an Enterococcus avium isolate and an Enterococcus faecium isolate from the post-heated fermented teas decreased the pH of the culture medium and increased the amount of lactic acid in the medium as they grew.DNeasy Blood & Tissue Kit by QIAGEN was used to extract DNA from the isolates. A primer was designed to multiply a 16S rRNA gene of about 1.5 kbp and used to analyze the extracted DNA. The analysis showed some differences in base sequence.A maze was used to judge the learning effect of y-amino butyric acid (GABA).Stress used in combination was also effective to judge the learning effect. A rat group given a forced administration of GABA 100 μg spent a longer time in threading the maze than a group given water. However, when the stress was used in combination, the GABA-administered group was able to thread the maze fastest.

在生产过程中对后加热的发酵茶进行取样，以研究从样品中分离出的微生物的特性。与 NBRC9405^T 不同，来自 Ishizuchi-kurocha 的真菌分离物没有假根。 NBRC31005 的孢子囊孢子形成片层。对分离真菌中的 28S rRNA 基因的 D2 区域进行测序，发现与变异根毛霉 (Rhizomucor variabilis) 的同源性为 98.7%。对来自 Awa-bancha、Ishizuchi-kurocha 和 Goishi-cha 的环状芽孢杆菌分离株进行了最适温度和 pH 值检测。最适温度为37-40℃，最适pH为6-7。发现后加热发酵茶的成分可以抑制分离株的生长。从后加热的发酵茶中分离出的鸟肠球菌和屎肠球菌在生长过程中都会降低培养基的 pH 值并增加培养基中的乳酸量。使用 QIAGEN 的 DNeasy 血液和组织试剂盒从分离物中提取 DNA。设计引物以扩增约1.5kbp的16S rRNA基因并用于分析提取的DNA。分析发现碱基序列存在一定差异。采用迷宫判断γ-氨基丁酸(GABA)的学习效果。组合使用压力也能有效判断学习效果。与给予水的组相比，强制给予 100 μg GABA 的大鼠组在迷宫中穿行的时间更长。然而，当压力结合使用时，GABA 给药组能够最快地穿过迷宫。