Acetone-butanol in-situ extraction continuous fermentation device and technology

A technology of acetone butanol and fermentation process, applied in fermentation, biochemical cleaning device, enzymology/microbiology device and other directions, can solve the problems of difficulty in large-scale production, high equipment price, poor stability, etc., and achieves easy implementation, The effect of increasing yield and reducing dosage

Inactive Publication Date: 2011-01-19

TIANJIN UNIVERSITY OF TECHNOLOGY

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Problems solved by technology

However, due to the poor stability and high equipment prices of these technologies in...

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Abstract

The invention provides an acetone-butanol in-situ extraction continuous fermentation device and technology. The device comprises an No.1 seeding tank and an No.2 seeding tank which are alternately inoculated. Acetone-butanol fermentation is divided into two stages to continuously operate, wherein the first stage is conventional acid production period fermentation, and the second stage is in-situ extraction product synthesis period fermentation; the second period adopts a method of coupling extraction and fermentation and is matched with low-speed stirring to furthest lower product inhibiting effect and greatly improve butanol fermentation efficiency; and finally, static phase processing realizes the initial separation of mash and fermentation products. The invention has the advantages that the device and the technology have good extraction effect in production, can obviously improve butanol ratio in the fermentation product, can greatly lower production cost, and have obvious economic benefit. The technology is scientific and reasonable. The flow device is simple and easy to realize, can realize the new fermentation technology provided by the invention by slightly reforming the traditional butanol fermentation flow, and has wide application prospect.

Application Domain

Bioreactor/fermenter combinationsBiological substance pretreatments +6

Technology Topic

ButanolChemistry +5

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Examples

Experimental program(2)
Effect test(1)

Example Embodiment

[0023] Example 1:

[0024] 1) Using corn steep liquor as raw material, its content in the culture medium is 20g/L, and additional nutrients are 0.3g/LCaCl 2 , 0.2g/L MgSO 4 ·7H 2 O, 0.5g/L KH 2 PO4, 0.5g/L urea; equipped with 0.5M NaOH injection lye tank; oleyl alcohol stock solution injected into the extractant storage tank for later use. The fermentation begins to steam sterilize the equipment in situ.

[0025] 2) Preparation of seed liquid: Clostridium acetobutylicun (CGMCCAS1.132) is used as the fermentation strain (purchased from the Common Microorganism Collection Center of the Chinese Microorganism Collection Management Committee), the composition of each liter of seed culture medium is: 40g glucose , 6g tryptone, 2g yeast extract, 2g beef extract, 3g (NH 4 ) 2 SO 4 , 0.2g MgSO 4 ·7H 2 O, 0.5g KH 2 PO 4 , 0.01g FeSO 4 ·7H 2 O, adjust pH=6.0~6.5. Inject the seed culture medium into the No. 1 seed fermentation tank, the injection volume is 2/3 of the volume of the fermentation tank, pass N 2 Deoxygenation for 2 to 3 hours. Use a concentration of 1×10 6 ~1×10 7 A cell/mL CGMCC AS1.132 bacterial suspension was connected to the No. 1 seed tank at 8% of the inoculum, and cultivated to the logarithmic growth phase under anaerobic conditions at 33°C; the No. 2 seed tank was seeded 8 hours after the same method The two seed tanks alternately cultivate the seed liquid every 6-8 hours.

[0026] 3) After the seed solution has been cultivated for 5 hours, turn on the raw material circulation pump of the first-level fermenter to make the raw material enter 2/3 of the volume of the fermenter, turn off the pump, and pass N 2 After deoxygenation, warm bath at 33°C for 2 to 3 hours. Connect the seed liquid in the No. 1 seed tank to the first-level fermentation tank at 5%, and incubate at 33°C for 8 hours to reach the logarithmic growth phase. Turn on the raw material circulation pump to circulate the feed, and the dilution rate is 0.03h -1; Simultaneously open the No. 2 seed tank inoculation valve, add seed liquid at an inoculum amount of 5% (v/v) per hour, continue the cultivation for 4 hours, overflow begins, and the overflow liquid enters the secondary fermentation tank. During continuous operation, the seed tank is switched every 7 hours.

[0027] 4) Feed 2/3 of the volume in the secondary fermentor in advance, and after heating at 33°C for 2 to 3 hours, pass N 2 Deaeration. The overflow liquid of the primary fermenter enters the secondary fermentor from the bottom, monitors the pH of the fermentation process, adds 0.5M NaOH dropwise to maintain the pH of the medium at 6.0-6.5, and starts stirring at low speed (125r/min). Add oleyl alcohol and raw materials at the same time, the ratio of oleyl alcohol to the medium added every hour is 1:6; the dilution rate of the added liquid is 0.03h -1 , Continuous cultivation for 8 hours, the overflow liquid enters the static phase tank.

[0028] Take the upper organic matter in the static phase tank for component analysis. The detection equipment used is Beijing Beifen Sp-34 gas chromatograph, the chromatographic column is capillary column 0V-17 (30m×0.25mm×0.25μm), and FID detector is used. The column temperature is 50°C, the temperature of the syringe and the detector are both 260°C. The initial temperature of the first program was 50°C, after maintaining it for 3 minutes, it rose to 160°C at a rate of 15°C/min, and the second time the temperature was programmed to increase from 20°C/min to 260°C for 2 minutes. Carrier gas is high purity N 2 , The carrier gas pressure is 0.12MPa. Table 1 shows the content of each component in the organic phase of continuous fermentation for 200 hours in Example 1.

[0029] Table 1 Composition of upper organic phase in static phase tank

[0030]

[0031] It can be seen from Table 1 that the upper organic phase in the static phase tank mainly contains fermentation products such as acetone butanol and the extractant oleyl alcohol, and the water content only accounts for 3.02%. After separating the extractant oleyl alcohol, the output of butanol accounted for 87.2% of the fermentation product output, which was an increase of 27% compared with the traditional fermentation process for CGMCC AS1.132 fermentation. The separated oleyl alcohol can be recycled.

Example Embodiment

[0032] Example 2:

[0033] 1) Using cassava pulp as raw material, the content in the medium is 40g/L, and the additional nutrients are 0.3g/LCaCl 2 , 0.2g/L MgSO 4 ·7H 2 O, 0.5g/L KH 2 PO4, 0.5g/L urea; equipped with 0.5M NaOH injection lye tank; oleyl alcohol stock solution injected into the extractant storage tank for later use. The fermentation begins to steam sterilize the equipment in situ.

[0034] 2) Preparation of seed liquid: Clostridium acetobutylicunm (CGMCCAS1.244) was used as the fermentation strain (purchased from the General Microorganism Collection Center of the China Microorganism Collection Management Committee).

[0035] The steps are the same as in Example 1, and the culture temperature is set to 35°C.

[0036] 3) After 7 hours of seed solution cultivation, turn on the raw material circulation pump of the first-level fermentation tank to make the raw materials enter 2/3 of the volume of the fermentation tank, turn off the pump, and connect 8% of the seeds in the first-level fermentation tank to the first-level fermentation tank Liquid, pass into N 2 After deoxygenation, culture at 35°C, monitor the pH of the fermentation process, and add 0.5M NaOH dropwise to maintain the pH of the medium at 6.0-6.5. After culturing for 12 hours, turn on the raw material circulation pump to circulate the feed, the dilution rate is 0.05h -1; At the same time, open the No. 2 seed tank inoculation valve, add seed liquid at an inoculum rate of 8% (v/v) per hour, continue to cultivate for 6 hours, overflow, and the overflow liquid enters the secondary fermentation tank. During continuous operation, the seed tank is switched every 8 hours.

[0037] 4) During secondary fermentation, cultivate at 35℃, the ratio of extractant to medium added every hour is 1:4; the dilution rate of added raw materials is 0.05h -1 , Continuously cultivate for 12 hours, and the overflow liquid enters the static phase tank.

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Broad application prospects
Easy to implement

Acetone-butanol in-situ extraction continuous fermentation device and technology

AI-Extracted Technical Summary

Problems solved by technology

Abstract

Image

Examples

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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