Bacillus subtilis and method for regulating and controlling molecular weight of fermentation product namely gamma-polyglutamic acid
A technology of Bacillus subtilis and polyglutamic acid, applied in the biological field, can solve the problems of low proportion of total polyglutamic acid, high loss rate of γ-polyglutamic acid, complicated process, etc. The effect of mitigating the damage of plants
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Embodiment 1
[0039] Utilize the fermentation of Bacillus subtilis XK producing γ-polyglutamic acid to produce γ-polyglutamic acid, the specific steps are as follows:
[0040] (1) Preparation of seed solution: Bacillus subtilis was inoculated with sodium glutamate (45 g / L), citric acid (50 g / L), bean cake powder (40 g / L), potassium dihydrogen phosphate (1 g / L) and water seed medium, pH (6.5), shaker speed 120~200rpm, culture at 28~36℃ for 10~18h to OD 660 Greater than 5.0;
[0041] (2) Fermentation culture: Inoculate the seed solution in step (1) with an inoculation amount of 3% v / v in a medium containing sodium glutamate (45 g / L), citric acid (50 g / L), soybean cake powder ( 40 g / L), potassium dihydrogen phosphate (1 g / L), manganese sulfate (0.05 g / L), magnesium sulfate (1.2g / L) in the fermentation medium, 7.5L fermenter fermentation medium liquid volume 4L, the aeration ratio was controlled at 1.0 vvm, the initial pH was 6.5, and cultured at 28 °C for 48 hours;
[0042] (3) The fermenta...
Embodiment 2
[0044] The preparation of the seed solution was the same as in Example 1.
[0045] (1) Fermentation culture: Inoculate the seed liquid into the fermentation medium with an inoculation amount of 4% v / v, which contains sodium glutamate (65 g / L), citric acid (34 g / L), bean cake Powder (20 g / L), potassium dihydrogen phosphate (1 g / L), manganese sulfate (0.09 g / L), magnesium sulfate (1.8 g / L) fermentation medium, 7.5L fermenter fermentation medium The liquid volume was 4L, the aeration ratio was controlled at 1.5 vvm, the initial pH was 6.9, and cultured at 32 °C for 52 h;
[0046] (2) The fermentation broth was diluted (concentration below 1 g / L), and then the molecular weight and yield of γ-polyglutamic acid in the fermentation broth were detected by high performance liquid chromatography. The molecular weight of the produced γ-polyglutamic acid was about 89 kDa, and the yield of γ-polyglutamic acid was 42.7 g / L.
Embodiment 3
[0048] The preparation of the seed solution was the same as in Example 1.
[0049] (1) Fermentation culture: Inoculate the seed liquid into the fermentation medium with an inoculation amount of 3% v / v, which contains sodium glutamate (51 g / L), citric acid (29 g / L), bean cake Powder (37 g / L), potassium dihydrogen phosphate (1 g / L), manganese sulfate (0.02 g / L), magnesium sulfate (1.1 g / L) fermentation medium, 7.5L fermenter fermentation medium The liquid volume was 4L, the aeration ratio was controlled at 1.2 vvm, the initial pH was 6.5, and cultured at 34 °C for 36 h;
[0050] (2) The fermentation broth was diluted (concentration below 1 g / L), and then the molecular weight and yield of γ-polyglutamic acid in the fermentation broth were detected by high performance liquid chromatography. The molecular weight of the produced γ-polyglutamic acid was about 38 kDa, and the yield of γ-polyglutamic acid was 36.8 g / L.
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