Bacillus subtilis producing gamma-poly(glutamic acid) and application thereof
A Bacillus subtilis, polyglutamic acid technology, applied in bacteria, microorganism-based methods, microorganisms, etc., can solve problems such as low yield, and achieve a simple process, suitable for industrialization promotion, good market prospects and economic benefits. Effect
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Embodiment 1
[0031] Utilize the fermentation of Bacillus subtilis XK producing γ-polyglutamic acid to produce γ-polyglutamic acid, the specific steps are as follows:
[0032] (1) Preparation of seed liquid: inoculate Bacillus subtilis into a mixture containing sodium glutamate (45g / L), citric acid (50g / L), bean cake powder (40g / L), potassium dihydrogen phosphate (1g / L) In the fermentation medium with water, the shaker speed is 120-200rpm, and cultured at 28-36°C for 10-18h to OD 660 Greater than 5.0;
[0033] (2) Fermentation culture: the seed liquid in the step (1) is inoculated with sodium glutamate (45g / L), citric acid (50g / L), bean cake powder (40g / L), In the fermentation medium of potassium dihydrogen phosphate (1g / L), manganese sulfate (0.05g / L), and magnesium sulfate (1.2g / L), the liquid volume of the fermentation medium in a 7.5L fermenter is 4L, and the aeration ratio is controlled at 1.0vvm, the initial pH is 6.5, cultured at 28°C for 48h;
[0034] (3) The fermentation broth i...
Embodiment 2
[0036] The preparation of the seed solution was the same as in Example 1.
[0037](1) Fermentation culture: the seed liquid is inoculated in the fermentation medium with 4% inoculation amount, contains sodium glutamate (65g / L), citric acid (34g / L), bean cake powder (20g / L) in the medium ), potassium dihydrogen phosphate (1g / L), manganese sulfate (0.09g / L), magnesium sulfate (1.8g / L) in the fermentation medium, the liquid volume of the 7.5L fermentation tank fermentation medium is 4L, and the aeration ratio Control at 1.5vvm, initial pH is 6.9, culture at 32°C for 52h;
[0038] (2) The fermentation broth is diluted (concentration is below 1g / L), and then the molecular weight and yield of γ-polyglutamic acid in the fermentation broth are detected by high performance liquid chromatography. The molecular weight of the produced γ-polyglutamic acid was about 89kDa, and the yield of γ-polyglutamic acid was 42.7g / L.
Embodiment 3
[0040] The preparation of the seed solution was the same as in Example 1.
[0041] (1) Fermentation culture: the seed liquid is inoculated in the fermentation medium with the inoculum of 3%, contains sodium glutamate (51g / L), citric acid (29g / L), bean cake powder (37g / L) in the medium ), potassium dihydrogen phosphate (1g / L), manganese sulfate (0.02g / L), magnesium sulfate (1.1g / L) in the fermentation medium, the liquid volume of the 7.5L fermentation tank fermentation medium is 4L, and the aeration ratio Controlled at 1.2vvm, the initial pH is 6.5, cultured at 34°C for 36h;
[0042] (2) The fermentation broth is diluted (concentration is below 1g / L), and then the molecular weight and yield of γ-polyglutamic acid in the fermentation broth are detected by high performance liquid chromatography. The molecular weight of the produced γ-polyglutamic acid is about 38kDa, and the yield of γ-polyglutamic acid is 36.8g / L.
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