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A method based on an electrochemical system to regulate intracellular reducing power regeneration and fermentation to produce succinic acid

A succinic acid production, electrochemical technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, fermentation, etc., can solve the problems of differences in electrochemical regulation methods of reducing power, etc.

Active Publication Date: 2018-11-23
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the reducing power required by different metabolites and the required electrochemical regulation methods are different.

Method used

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  • A method based on an electrochemical system to regulate intracellular reducing power regeneration and fermentation to produce succinic acid
  • A method based on an electrochemical system to regulate intracellular reducing power regeneration and fermentation to produce succinic acid
  • A method based on an electrochemical system to regulate intracellular reducing power regeneration and fermentation to produce succinic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] This example illustrates that Escherichia coli ( Escherichia coli ) AFP111 method for anaerobic fermentation.

[0044] Escherichia coli( Escherichia coli ) The AFP111 anaerobic fermentation method is as follows:

[0045] The Escherichia coli in the cryopreservation tube ( Escherichia coli ) AFP111 as the starting strain was inoculated with 1% (v / v) inoculum in a test tube containing 5mL seed culture medium, cultivated at 37°C, 200r / min for 12h to obtain the first-level seed solution; %(V / v) inoculum was inoculated into a 500mL triangular shake flask containing 100mL seed culture medium, cultured at 37°C and 200r / min for 6h to obtain a secondary seed solution. The secondary seed liquid was inoculated into a 700mL chemostat device containing 450mL of fermentation medium at an inoculum of 10% (v / v). At the same time, sterile carbon dioxide was continuously introduced into the chemostat to maintain anaerobic surroundings. Regularly aseptic samples are taken during the fermenta...

Embodiment 2

[0051] This example illustrates the use of Escherichia coli under the condition of -0.65V and the addition of 0.05mmol / L neutral red ( Escherichia coli ) AFP111 is a one-step electrochemical anaerobic fermentation method.

[0052] Escherichia coli( Escherichia coli ) The one-step electrochemical anaerobic fermentation method of AFP111 is as follows:

[0053] The Escherichia coli in the cryopreservation tube ( Escherichia coli ) AFP111 as the starting strain was inoculated with 1% (v / v) inoculum in a test tube containing 5mL seed culture medium, cultivated at 37°C, 200r / min for 12h to obtain the first-level seed solution; %(V / v) inoculum was inoculated into a 500mL triangular shake flask containing 100mL seed culture medium, cultured at 37°C and 200r / min for 6h to obtain a secondary seed solution. The secondary seed liquid was inoculated into a 700mL cathode chamber containing 450mL of fermentation medium at an inoculum of 10% (v / v). At the same time, a voltage of -0.65V was applied...

Embodiment 3

[0059] This example illustrates the use of Escherichia coli under the condition of -0.65V and 0.1mmol / L riboflavin ( Escherichia coli ) AFP111 is a one-step electrochemical anaerobic fermentation method.

[0060] Escherichia coli( Escherichia coli ) The one-step electrochemical anaerobic fermentation method of AFP111 is as follows:

[0061] The Escherichia coli in the cryopreservation tube ( Escherichia coli ) AFP111 as the starting strain was inoculated with 1% (v / v) inoculum in a test tube containing 5mL seed culture medium, cultivated at 37°C, 200r / min for 12h to obtain the first-level seed solution; %(V / v) inoculum was inoculated into a 500mL triangular shake flask containing 100mL seed culture medium, cultured at 37°C and 200r / min for 6h to obtain a secondary seed solution. The secondary seed liquid was inoculated into a 700mL cathode chamber containing 450mL of fermentation medium at an inoculum of 10% (v / v). At the same time, a voltage of -0.21V was applied to the cathode ch...

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Abstract

The invention discloses a method for fermented production of succinic acid on the basis of an electrochemical system for regulating intracellular reducing power regeneration. The method comprises three steps including strain activation, seed culture and anaerobic fermentation production of succinic acid, anaerobic fermentation adopts electrochemical fermentation, an electron carrier is added to a fermentation medium, the corresponding voltage is applied to a cathode, and the concentration of the electron carrier is in a range of 0.01-0.5 mmol / L. the electron carrier is neutral red or riboflavin. The cathode voltage applied to the cathode is scanned and determined in the range from 0.1 V to 1 V with the cyclic voltammetry at the scanning speed of 5-10 mV / s. When fermentation is performed in an electrochemical device under the anaerobic condition, the total amount of intracellular NADH is increased under the assistance of the electron carrier, the intracellular reducing power (NADH / NAD+) level is tripled, the succinic acid accumulation reaches 15.06 g / L, and efficient synthesis of the reduced product, namely, the succinic acid, is facilitated.

Description

Technical field [0001] The invention relates to a method for regeneratively fermented to produce succinic acid based on an electrochemical system regulating the reducing power in a cell, and belongs to the technical field of biochemical industry. Background technique [0002] For different carbon sources and products, the reducing power supply and consumption levels are not always the same. For example, when 1 molecule of glucose and 1 molecule of glycerol are converted into phosphoenolpyruvate via glycolysis, 2 molecules of NADH are produced. If the final product is succinic acid or ethanol with strong reducing properties, when glucose is the only carbon source, the reducing power for product synthesis is obviously insufficient, which reduces the yield of reducing products, and when glycerol is used as the carbon source , And the growth of bacteria will be stagnant due to excess reducing power. In order to balance the metabolism of intracellular coenzymes and restore the growth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/46C12R1/19
CPCC12P7/46
Inventor 姜岷冀亚亮马江锋吴明科吴昊张敏
Owner NANJING TECH UNIV
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