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Method for increasing butanol-acetone ratio and butanol yield of clostridia ABE fermentation

A technology of Clostridium acetobutylicum and butanol fermentation, which is applied in the field of ABE fermentation, can solve the problems of increasing difficulty, butanol production does not reach the inhibitory concentration of Clostridium acetobutylicum, and the ratio of butanol/acetone does not increase significantly, so as to improve the comprehensive performance , simple implementation, effective use of waste water

Active Publication Date: 2015-02-25
JIANGNAN UNIV
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  • Description
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AI Technical Summary

Problems solved by technology

However, there are obvious defects in the above methods of improving butanol / acetone ratio or butanol output: either increase the difficulty of subsequent separation and purification (removing pigment substances, etc.), or sacrifice the total solvent output
However, although the existing mixed culture mode has improved the fermentation performance of ABE to a certain extent, the butanol production has not reached the inhibitory concentration of Clostridium acetobutylicum (about 13g / L), and the butanol / acetone ratio has not increased significantly.

Method used

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  • Method for increasing butanol-acetone ratio and butanol yield of clostridia ABE fermentation
  • Method for increasing butanol-acetone ratio and butanol yield of clostridia ABE fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 corn butanol traditional fermentation

[0023] Strain activation: inoculate Clostridium acetobutylicum ATCC824 stored in a refrigerator at 4°C in 5% (w / v) corn mash seed medium, remove the air with a vacuum pump, and culture it statically in a water bath at 37°C for 27 hours as Fermented inoculum. Wherein, the preparation process of corn mash seed culture medium is as follows: take 25g corn flour and dissolve in 2L water, gelatinize in boiling water at high temperature until the volume of corn mash solution is 500mL, which is 5% (w / v) corn seed Culture medium, the sterilization condition is 121°C, 50min.

[0024] The fermentation medium is a dual-enzyme hydrolyzate of 15% (w / v) corn flour, and the preparation process is as follows: the first step is to add high-temperature amylase in an environment of 95°C, the addition amount is 8U / g-starch, and liquefy for 45min; Cool the enzymatic hydrolyzate obtained in the first step of hydrolysis to 62°C, add glucoa...

Embodiment 2

[0030] Embodiment 2 single adding Saccharomyces cerevisiae

[0031] In Example 2, the strain activation and fermentation conditions were the same as in Example 1. When the fermentation was carried out to 24h, the pH of the butanol fermentation broth was adjusted to 5.0 with 3mol / L sodium hydroxide solution, and then 10 % (v / v) of Saccharomyces cerevisiae broth until the end of fermentation. The medium formula of Saccharomyces cerevisiae is: glucose 20g / L, yeast extract 8.5g / L, ammonium sulfate 1.3g / L, magnesium sulfate heptahydrate 0.1g / L, calcium chloride dihydrate 0.06g / L, sterilized The conditions were 121°C, 20min; at the beginning of ABE fermentation, the Saccharomyces cerevisiae slant stored in the refrigerator at 4°C was transferred to a 500mL Erlenmeyer flask containing 100mL yeast medium, cultured at 30°C, 200r / min for 24h, then When entering the ABE fermentation broth, the yeast culture broth OD 600 8.5-9.5, pH 4.4-4.8.

[0032] The operating conditions of the ana...

Embodiment 3

[0034] Embodiment 3 single addition of butyric acid

[0035] The strain activation and fermentation conditions in Example 3 were the same as in Example 1, and when the fermentation was carried out to 24 hours, the butyric acid solution (pH 3.8) was added in pulses. An organic acid was added every 3 hours until the pH dropped by 0.1 each time. And record the balance readings before and after the addition to calculate the amount of organic acid added. The addition process is accompanied by slight stirring (the rotation speed of the stirring paddle is 100r / min), so that the added organic acid solution and the fermentation broth are mixed quickly to ensure that the pH of each part of the fermentation broth is uniform. After the addition process, the stirring is turned off. The total amount of butyric acid added is 4g / L.

[0036] The operating conditions of the fermenter were the same as in Example 1. The mensuration of parameter: the mensuration of relevant parameter is the sa...

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Abstract

The invention discloses a method for increasing the butanol-acetone ratio and the butanol yield of clostridia ABE fermentation, and belongs to the technical field of biology. According to the method, corn flour is taken as a fermentation raw material, and a clostridium-saccharomyces cerevisiae mixed culture system is formed by appropriately adding an active saccharomyces cerevisiae and a butyric acid solution at the solvent production stage of the clostridia. The saccharomyces cerevisiae is capable of secreting various amino acids under a strict anaerobic condition and in a severe saccharomyces cerevisiae growth environment, wherein aromatic amino acids and aspartic amino acids all advantageous for butanol synthesis are greatly accumulated. On the basis, the small amount of added butyric acid is further capable of weakening the metabolism intensity of the closed loop of butyric acid. Under the dual combined action, the butanol-acetone ratio and the final butanol concentration of the ABE fermentation can be greatly improved under the premise of unchanged or slightly improved total solvent production intensity. The two indexes, namely butanol-acetone ratio and butanol concentration of the method are at the highest level in the related study reports of the batch fermentation production of butanol by use of wild clostridia and have important guiding significance for the ABE fermentation.

Description

technical field [0001] The invention relates to a method for improving the butanol / acetone ratio and butanol yield of ABE fermentation, especially an ABE fermentation method for improving the butanol / acetone ratio and butanol yield through mixed culture of Clostridium acetobutylicum and Saccharomyces cerevisiae, belonging to biotechnology field. Background technique [0002] Butanol is an important platform compound, which is widely used in chemical industry, plastics, medicine, organic synthesis, printing and dyeing and other industries. At the same time, bio-butanol is also a promising, clean and renewable new liquid fuel that can be used as a substitute for fossil fuels. The development of biofuels has become an important measure for countries all over the world to deal with the energy crisis, reduce greenhouse gas emissions, and improve the climate environment. Butanol fermentation is generally divided into two stages: the first stage is the acidogenic stage, butyric a...

Claims

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Application Information

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IPC IPC(8): C12P7/16C12R1/45
CPCC12P7/16Y02E50/10
Inventor 史仲平罗洪镇李志刚何珍妮陈锐
Owner JIANGNAN UNIV
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