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

在生产过程中对加热后的发酵茶进行取样，以研究从样品中分离出的微生物的性质。与NBRC9405^T不同，从Ishizuchi-kurocha分离的真菌没有根状体。NBRC31005的孢子囊孢子形成片层。对该真菌分离物28S rRNA基因D2区进行测序，发现与变异根霉基因D2区同源性为98.7%。对阿和班茶、石井黑茶和Goishi-cha 3株圆形芽孢杆菌进行了最适温度和pH的测定，最适温度为37 ~ 40℃，最适pH为6 ~ 7。经热后发酵茶的成分对菌株的生长有抑制作用。从加热后的发酵茶中分离出的鸟肠球菌和屎肠球菌都降低了培养基的pH值，并增加了培养基中乳酸的含量。采用QIAGEN公司的DNeasy血液和组织试剂盒提取分离株的DNA。设计引物扩增约1.5 kbp的16S rRNA基因，用于分析提取的DNA。分析结果显示碱基序列存在一定差异。采用迷宫法评价氨基丁酸（GABA）的学习效果。压力组合使用对判断学习效果也是有效的。被强制给予100 μg GABA的大鼠组比被给予水的大鼠组穿过迷宫的时间更长。然而，当压力组合使用时，gaba组能够最快地穿过迷宫。

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.