Method for producing succinic acid by utilizing Escherichia coli BA305 through fermentation
A technology of Escherichia coli and succinic acid, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problem of the decrease of yield and production intensity, the decrease of bacterial body density, the decrease of specific production intensity of cells, Insufficient supply of ATP and other problems, to reduce costs and increase the effect of practical scope
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Embodiment 1
[0073] This example illustrates the comparison of Escherichia coli BA305, the starting strain NZN111 and the control strain AFP111 in the production of succinic acid by recycling glucose fermentation.
[0074] First use LB medium to activate the thalli, inoculate Escherichia coli BA305 into LB medium with a liquid volume of 3L in a 7.5L fermenter according to 1% (v / v) inoculum, and add 15 g / L glucose to 0.5 L / min speed into the air or pure oxygen, when aerobic culture bacteria OD 600 Induced with 0.3 mM IPTG to 0.4, cultured aerobically for 15h to OD 600 =19, stop feeding air or pure oxygen, and feed CO at a rate of 0.5 L / min 2 Anaerobic fermentation is carried out to produce acid, and the maximum concentration of glucose added does not exceed 30 g / L. At the same time, a neutralizing agent basic magnesium carbonate is added to adjust the pH value to 6.4. When the glucose consumption rate of the cells drops to 0.6 g / (L h), the glucose content in the culture medium is detected...
Embodiment 2
[0086] This example illustrates the comparison of Escherichia coli BA305, the starting strain NZN111 and the control strain AFP111 in the production of succinic acid by recycling xylose fermentation.
[0087] First use LB medium to activate the thalline, Escherichia coli BA305 is inoculated into 7.5L fermenter tank liquid volume is 3L LB medium and adds 15 g / L xylose by 1% (v / v) inoculum amount, Infuse air or pure oxygen at a rate of 0.5 L / min, when the OD of aerobic culture cells 600 Induced with 0.3 mM IPTG to 0.6, cultured aerobically for 15h to OD 600 =19, stop feeding air or pure oxygen, and feed CO at a rate of 0.5 L / min 2 Perform anaerobic fermentation to produce acid, add xylose at a maximum concentration of no more than 30 g / L, and add sodium carbonate to adjust the pH to 6.8. When the xylose consumption rate of the cells drops to 0.6 g / (L h), the xylose content in the medium is detected, and the bacterial cells are obtained by aseptic centrifugation, and the concen...
Embodiment 3
[0099] This example illustrates the comparison of Escherichia coli BA305, the starting strain NZN111 and the control strain AFP111 in the production of succinic acid by recycling fructose fermentation.
[0100] First use LB medium to activate the thalli, inoculate Escherichia coli BA305 into 7.5L fermenter with 3L LB medium and add 15 g / L fructose according to 1% (v / v) inoculum amount, to 0.5 L / min speed into the air or pure oxygen, when aerobic culture bacteria OD 600 Induced with 0.3 mM IPTG to 0.5, cultured aerobically for 15h to OD 600 =19, stop feeding air or pure oxygen, and feed CO at a rate of 0.5 L / min 2 Perform anaerobic fermentation to produce acid, add fructose at a maximum concentration of no more than 30 g / L, and add sodium bicarbonate to control the pH value to 6.6. When the fructose consumption rate of the cells drops to 0.6 g / (L h), the fructose content in the culture medium is detected, and the bacterial cells are obtained by aseptic centrifugation, and the...
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