Screening of Enterococcus faecium Producing γ-Amminobutyric Acid and Quantitative Detection of γ-Amminobutyric Acid

ZHU Quan; CHENG Jinlong; ZHU Yuanzhao; YIN Long; NI Jindong; CHENG Maoji; YANG Zhangping

doi:10.3969/j.issn.1006-267x.2016.08.022

Chinese Journal of Animal Nutrition >

2016 , Vol. 28 >Issue 8: 2504 - 2511

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.08.022

FEED SCIENCE AND TECHNOLOGY

Screening of Enterococcus faecium Producing γ-Amminobutyric Acid and Quantitative Detection of γ-Amminobutyric Acid

ZHU Quan ,
CHENG Jinlong ,
ZHU Yuanzhao ,
YIN Long ,
NI Jindong ,
CHENG Maoji ,
YANG Zhangping

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1. College of Animal Science, Anhui Agricultural University, Hefei 230061, China;
2. Jiangsu Unison Biotechnology Development Co., Ltd., Suqian 223831, China;
3. College of Animal Science, Anhui Science and Technology University, Fengyang 233100, China;
4. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

Received date: 2016-02-24

Online published: 2016-08-17

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Abstract

This study was aimed to isolate and identify an Enterococcus faecium by molecular biological methods which produced γ-aminobutyric acid (GABA), moreover, the amount of GABA was also quantitatively determined. One target strain of Enterococcus faecium F6 was screened out from the samples of pickles, yogurt, soil and fresh milk, and was identified according the morphological characters and Gram stain methods. After the 16S rDNA gene of bacterial strain F6 was amplified, the gene sequence and phylogenetic tree were analyzed. Meanwhile, the GABA content in the fermented solution of bacterial strain F6 was quantitatively determined by high performance liquid chromatography (HPLC) method. The results showed that the bacterial strain F6 took a large, smooth, round and a neat edge of shape with a diameter of 1 to 2 mm and in a milky white color. A transparent circle was formed around the colony on the separating medium which changed to be yellow. On the MRS solid medium, the colony was opaque with a transparent circle around. Gram stain was used to identify F6 as a positive strain. The 16S rDNA gene sequence of bacterial strain F6 was more than 99% similarity to that of Enterococcus faecium in GenBank showed by molecular biological analysis. HPLC method was used to draw out the GABA standard curve linear equation as follows:Y=7 080 733.139 5X-4 511.692 7 (R²=0.999 4), and the content of 7.1 g/L of GABA with 7 to 10 minutes of retention time was determined in the fermented solution of F6 according to linear equation. The results indicate that a high yield GABA of Enterococcus faecium F6 is finally obtained.

Key words： Enterococcus faecium; γ-aminobutyric acid; 16S rDNA sequence; quantitative detection by high performance liquid chromatography method; screening

Cite this article

ZHU Quan , CHENG Jinlong , ZHU Yuanzhao , YIN Long , NI Jindong , CHENG Maoji , YANG Zhangping . Screening of Enterococcus faecium Producing γ-Amminobutyric Acid and Quantitative Detection of γ-Amminobutyric Acid[J]. Chinese Journal of Animal Nutrition, 2016 , 28(8) : 2504 -2511 . DOI: 10.3969/j.issn.1006-267x.2016.08.022

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