Lactobacillus fermentum capable of producing gamma-aminobutyric acid with high yield and application thereof
A technology of fermenting lactobacillus and aminobutyric acid, applied in the field of natural strains, to achieve good adaptability
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example 1
[0016] The screening method of the lactobacillus fermentum of example 1 high production gamma-aminobutyric acid
[0017] The lactic acid bacteria preserved on the MRS agar plate were inoculated in the fermentation medium containing 1% glutamic acid for 72 hours after secondary activation in the MRS liquid medium, and 1-2 μl of the fermentation liquid was taken and analyzed by paper chromatography. Preliminary screening of γ-aminobutyric acid producing bacteria.
[0018] The developing agent used for paper chromatography in the present invention is: V (n-butanol): V (glacial acetic acid): V (water)=4:2:1, and 0.4% ninhydrin. A 5% γ-aminobutyric acid and 5% glutamic acid solution was used as a control. The chromatography time is 4-6 hours. After the chromatography is finished, dry the filter paper to see whether the fermentation broth shows purple-red spots at the corresponding sites of the γ-aminobutyric acid standard sample.
[0019] According to the preliminary screening re...
example 2
[0023] Example 2 shake flask fermentation production gamma-aminobutyric acid
[0024] The Lactobacillus fermentum preserved on the MRS agar plate was inoculated into the shake flask fermentation medium with a 10% inoculation amount after secondary activation in the MRS liquid medium, at a rotation speed of 200 rpm, and cultured at 30°C for 48 hours. The γ-amino group in the fermentation liquid The content of butyric acid is about 12 g / l. The formula of shake flask fermentation medium is: 1% peptone, 0.5% yeast extract, 1% beef extract, 2% glucose, 5% sodium glutamate monohydrate, 0.3% anhydrous sodium acetate, 0.2% triscitric acid Ammonium, 0.2% dipotassium hydrogen phosphate, 0.01% anhydrous magnesium sulfate, 0.025% manganese sulfate monohydrate, 0.1% Tween 80. The detection method of γ-aminobutyric acid is the same as that of Example 1, and the output of γ-aminobutyric acid is about 12 g / l.
example 3
[0025] Example 3 Static fermentation produces γ-aminobutyric acid
[0026]The Lactobacillus fermentum preserved on the MRS agar plate was inoculated in the static fermentation medium with an inoculum of 10% after secondary activation in the MRS liquid medium, and fermented statically at 30°C for 96 hours. The content of γ-aminobutyric acid in the fermentation liquid The content is about 38g / l. The formula of static fermentation medium is: 1.5% soybean peptone, 3% yeast extract, 3% glucose, 8% sodium glutamate monohydrate, 0.3% anhydrous sodium acetate, 0.2% triammonium citrate, 0.2% dihydrogen phosphate Potassium, 0.01% magnesium sulfate anhydrous, 0.025% manganese sulfate monohydrate, 0.1% Tween 80. The detection method of γ-aminobutyric acid is the same as that of Example 1, and the output of γ-aminobutyric acid is about 38g / l.
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