A high-yielding gamma-polyglutamic acid strain and a method for preparing gamma-polyglutamic acid by liquid fermentation using the strain
A technology of polyglutamic acid and high-yielding strains is applied in the field of bioengineering to achieve the effects of improving fertilizer efficiency, improving disease resistance and reducing environmental pressure
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Embodiment 1
[0088] A method for preparing gamma-polyglutamic acid by liquid fermentation, the specific steps are as follows:
[0089] (1) Activation of strains: move the strains on the slant of the test tube stored at 4°C on the LB medium to room temperature for activation for 4 hours;
[0090] (2) Preparation of liquid seeds: On an aseptic operating table, use a sterile inoculation loop to inoculate 2 single colonies into a triangular flask containing sterilized seed medium, culture at 30°C, shaker speed at 170rpm, and culture for 24 hours ;
[0091] Seed medium composition: glucose 20g / L, yeast extract 7g / L, sodium glutamate 19g / L, K 2 HPO 4 ·3H 2O 2g / L, MgSO 4 0.20g / L, distilled water 1000mL, pH 7.3, sterilization conditions 121°C, 0.15Mpa, sterilize for 20min.
[0092] (3) Fermentation, regulation and control process: the liquid seed is received in the fermentation medium with the inoculation amount of 7% (v / v), and the 500L fermenter fermentation medium filling liquid volume is...
Embodiment 2
[0100] A method for preparing gamma-polyglutamic acid by liquid fermentation, the specific steps are as follows:
[0101] (1) Activation of strains: move the strains on the slant of the test tube stored at 4°C on the LB medium to room temperature for activation for 4 hours;
[0102] (2) Preparation of liquid seeds: On an aseptic operating table, use a sterile inoculation loop to inoculate 2 single colonies into a triangular flask containing sterilized seed medium, cultivate at a temperature of 32°C, rotate at a shaker speed of 180rpm, and cultivate for 20 hours ;
[0103] Seed medium composition: glucose 20g / L, yeast extract 8g / L, sodium glutamate 20g / L K 2 HPO 4 ·3H 2 O 2g / L, MgSO 4 0.20g / L, distilled water 1000mL, pH 7.5, sterilization condition 121℃, 0.15Mpa, sterilization 20min;
[0104] (3) Fermentation, regulation and control process: the liquid seed is received in the fermentation medium with the inoculum size of 5% (v / v), and the liquid filling capacity of the fe...
Embodiment 3
[0112] A method for preparing gamma-polyglutamic acid by liquid fermentation, the specific steps are as follows:
[0113] (1) Activation of strains: move the strains on the slant of the test tube stored at 4°C on the LB medium to room temperature for activation for 4 hours;
[0114] (2) Preparation of liquid seeds: On an aseptic operating table, use a sterile inoculation loop to inoculate 3 single colonies into a triangular flask containing sterilized seed medium, cultivate at a temperature of 35°C, rotate at a shaker speed of 190rpm, and cultivate for 16 hours ;
[0115] Seed medium composition: glucose 18g / L, yeast extract 8g / L, sodium glutamate 20g / L K 2 HPO 4 ·3H 2 O 2g / L, MgSO 4 0.20g / L, distilled water 1000mL, pH 7.0, sterilization condition 121℃, 0.15Mpa, sterilization 20min;
[0116] (3) Fermentation and regulation process: the liquid seed is received in the fermentation medium with the inoculum size of 5% (v / v), and the liquid filling capacity of the fermentatio...
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