High-yield production method of GABA (gamma amino butyric acid) through mixed fermentation of brevibacterium tianjinese and lactobacillus plantarum

A technology of Lactobacillus plantarum and Brevibacterium tianjini, applied in the fields of fermentation engineering and enzyme engineering, can solve problems such as lack of γ-aminobutyric acid synthesis pathway, and achieve the effects of improving production efficiency, reducing production cost and improving safety

Active Publication Date: 2013-08-14
JIANGNAN UNIV
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Problems solved by technology

The strain lacks the GABA synthesis pathway, but can efficiently convert glucose to glutamate

Method used

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  • High-yield production method of GABA (gamma amino butyric acid) through mixed fermentation of brevibacterium tianjinese and lactobacillus plantarum
  • High-yield production method of GABA (gamma amino butyric acid) through mixed fermentation of brevibacterium tianjinese and lactobacillus plantarum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: culture medium, is described as follows:

[0021] (1) Brevibacterium tianjin SW07-1

[0022] ①Activation medium: peptone 1, yeast extract 0.5, NaCl1, glucose 0.5, sterilized at 115°C for 15 minutes; ②Seed medium: glucose 30, corn steep liquor 25, KH 2 PO 4 ·3H 2 O1.5, MgSO 4 ·7H 2 O0.4, urea 6 (diluted), pH7.0-7.2, sterilized at 115°C for 15 minutes; ③Fermentation medium (g / L): glucose 140, corn steep liquor 3, urea 5.5 (dissolved), KH 2 PO 4 ·3H 2 O1.5, MgSO 4 ·7H 2 O0.8, MnSO 4 ·H 2 O0.02, FeSO 4 ·7H 2 O0.02, biotin 8×10 -5 , L-histidine 5×10 -4 , pH7.0-7.2, sterilized at 115°C for 15min.

[0023] (2) L. plantarum GB01-21

[0024]①Activation medium (g / L): MRS medium. Casein peptone 10, beef extract 10, yeast extract 5, glucose 5, sodium acetate 5, diamine citrate 0.2, Tween 0.1, dipotassium hydrogen phosphate 0.2, magnesium sulfate 0.2, manganese sulfate 0.05, calcium carbonate 20, pH6 .5, sterilized at 115°C for 15 minutes; ②Seed mediu...

Embodiment 2

[0025] Example 2: Glutamic acid fermentation of Brevibacterium tianjin SW07-1 and collection of L.plantarum GB01-21

[0026] Cultivation of the glutamic acid-producing strain Brevibacterium tianjin SW07-1: Inoculate the preserved strain into the activation medium, 30°C, 220r / min reciprocating shaker shake culture for 12h, and then inoculate the seed medium with 2% inoculum , 30°C, 220r / min reciprocating shaker for 12h. Then inoculate into 5L fermenter with 10% inoculum amount. The liquid volume in the fermenter is 2.5L, the ventilation rate is 3.5L / min, the pH is controlled by feeding ammonia water to control pH7.0-7.2, the speed is adjusted to 400-600r / min according to the dissolved oxygen demand, and the culture is carried out at 30°C for 36h. By adding an appropriate amount of glucose, the final glutamic acid content in the fermentation broth is about 90g / L.

[0027] Collection of Lactobacillus plantarum: Inoculate the preserved strains into the activated medium MRS mediu...

Embodiment 3

[0028] Embodiment 3: direct conversion of glutamic acid fermentation broth

[0029] Add the corresponding amount of anhydrous sodium acetate and glacial acetic acid to Brevibacterium tianjin SW07-1 glutamic acid fermentation liquid according to the preparation method of acetic acid buffer to form a 0.2mol / L, pH 5.0 acetic acid buffer system. At the same time, add the plantaractobacillus collected in the previous step into the buffer system (the amount of added thalli is calculated according to the volume of plantaractobacillus culture solution: the volume of glutamic acid fermentation solution = 3: 2), as the transformation system, the stirring speed and aeration rate were 150r / min and 0.5L / min, respectively, and transformed at 30°C, and the residual glutamic acid content in the fermentation broth was regularly detected. like figure 1 Shown is the change curve of the original glutamic acid content in the glutamic acid fermentation broth during the conversion process, and the ...

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Abstract

The invention discloses a high-yield production method of GABA (gamma amino butyric acid) through mixed fermentation of brevibacterium tianjinese and lactobacillus plantarum, and belongs to the fields of fermentation engineering and enzyme engineering. According to the invention, glucose is transformed into glutamic acid by using the brevibacterium tianjinese SW07-1, and then glutamic acid of fermentation liquor obtained in the last step is transformed into GABA by using the lactobacillus plantarum GB01-21. Before transformation, the pH value of glutamic acid fermentation liquor is adjusted to be the optimum pH value of glutamic acid decarboxylase, so that a transformation system is built, and in the step 2, after the fermentation liquor is filtrated or centrifuged, the lactobacillus plantarum GB01-21 is obtained, and then the lactobacillus plantarum GB01-21 is added into the transformation system to carry out whole-cell transformation. When original glutamic acid of the fermentation liquor is completely transformed, GABA in the transformation liquor is 59.2 g/L, and the molar transformation rate is 93.6%. After an exogenous glutamic acid is continued to be added and then transformed, the GABA content is 96.5 g/L, and the molar transformation rate is 91.8%.

Description

technical field [0001] The invention discloses a method for high-yield GABA mixed fermentation by using safe and efficient Brevibacterium tianjin and Lactobacillus plantarum, belonging to the fields of fermentation engineering and enzyme engineering. technical background [0002] γ-Aminobutyric acid (GABA) is a non-protein amino acid naturally present in some organisms. It is an important inhibitory neurotransmitter in the central nervous system of mammals and has a wide range of applications in food, feed, medicine and other fields. γ-aminobutyric acid has physiological functions such as anti-anxiety, lowering blood pressure, calming the nerves, enhancing memory, regulating hormone secretion, promoting reproduction, diuresis, and analgesia. [0003] The preparation of γ-aminobutyric acid is mainly through chemical synthesis and biological methods. The biological method includes plant enrichment method and microbial fermentation method. The chemical method is mainly obtain...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12R1/25C12R1/13
Inventor 饶志明孙红梅李秀鹏徐美娟
Owner JIANGNAN UNIV
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