A method for producing glucosamine by fermentation
A technology of glucosamine and fermentation medium, applied in microorganism-based methods, biochemical equipment and methods, fermentation and other directions, can solve the problems of low fermentation density of strains, low glucosamine production, affecting the extracellular accumulation of glucosamine, etc. , to achieve the effect of increasing production
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Embodiment 1
[0055] Specific steps are as follows:
[0056] (1) Put 50mL seed medium in a 250mL shake flask, sterilize at 121°C for 20min, pick an agar block with a diameter of 1cm from the seed slope of Escherichia coli BL21 and inoculate it into the medium, 36°C, 200rpm Cultivate for 10 hours to obtain seed solution;
[0057] (2) Add 2g / L pyruvic acid, 4.5g / L aspartic acid, 0.3g / L inositol in the fermentation medium;
[0058] (3) Put 100mL fermentation medium containing 2g / L pyruvate, 4.5g / L aspartic acid, and 0.3g / L inositol into a 500nL shaker flask, sterilize at 121°C for 20min, and use 10% inoculum The seed solution was inoculated into the medium, and cultured at 36° C. and 280 rpm for 40 hours to obtain a fermentation solution.
[0059] Detect the output of glucosamine in the fermentation broth, and the output is 59g / L.
Embodiment 2
[0061] Specific steps are as follows:
[0062] (1) Put 50mL seed medium in a 250mL shake flask, sterilize at 121°C for 20min, pick an agar block with a diameter of 1cm from the seed slope of Escherichia coli BL21 and inoculate it into the medium, 36°C, 200rpm Cultivate for 10 hours to obtain seed solution;
[0063] (2) Add 2g / L pyruvate, 4.5g / L serine, 0.3g / L inositol to the fermentation medium;
[0064] (3) Put 100mL fermentation medium containing 2g / L pyruvate, 4.5g / L serine, and 0.3g / L inositol in a 500nL shake flask, sterilize at 121°C for 20min, and inoculate the seed solution with 10% inoculum Inoculate into the culture medium, culture at 36° C. and 280 rpm for 40 hours to obtain a fermentation broth.
[0065] Detect the output of glucosamine in the fermentation broth, and the output is 52g / L.
Embodiment 3
[0067] Specific steps are as follows:
[0068] (1) Put 50mL seed medium in a 250mL shake flask, sterilize at 121°C for 20min, pick an agar block with a diameter of 1cm from the seed slope of Escherichia coli BL21 and inoculate it into the medium, 36°C, 200rpm Cultivate for 10 hours to obtain seed solution;
[0069] (2) Add 1g / L pyruvate, 3g / L aspartic acid, 0.2g / L inositol to the fermentation medium;
[0070] (3) Put 100mL fermentation medium containing 1g / L pyruvate, 3g / L aspartic acid, and 0.2g / L inositol in a 500nL shake flask, sterilize at 121°C for 20min, and inoculum with 10% inoculum The seed solution was inoculated into the culture medium, and cultured at 36° C. and 280 rpm for 40 hours to obtain a fermentation broth.
[0071] Detect the output of glucosamine in the fermentation broth, and the output is 46g / L.
PUM
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