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Clostridium acetobutylicum and applications thereof

A technology of Clostridium acetobutylicum and butanol, which is applied in the field of microorganisms, can solve the problems such as the effective utilization of NADH and the waste of NADH in the separate regulation of difficult metabolic flow, and achieve flexible adjustment of metabolic flow, increase carbon yield, and improve product yield. Effect

Active Publication Date: 2013-09-25
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Bacillus subtilis usually grows or ferments aerobically, and because each molecule of glucose to 3-hydroxybutanone will produce 2 NADH, the exclusive production of 3-hydroxybutanone will cause waste of NADH
At the same time, since 2,3-butanediol and 3-hydroxybutanone are in the upstream and downstream of the same branch, it is difficult to achieve independent regulation of the metabolic flux of the two and the effective utilization of NADH by conventional co-production

Method used

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  • Clostridium acetobutylicum and applications thereof
  • Clostridium acetobutylicum and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Mutagenesis of Clostridium acetobutylicum

[0052] Clostridium acetobutylicum (Clostridium acetobutylicum) was used as the original bacterial strain, activated and cultured on the plate for 24 hours, then transferred once, and a ring of bacteria sludge was picked and added to 60mM lithium chloride sterile aqueous solution with 100 glass beads. Shake 100ml in a shaker flask at 200rpm for 10min to evenly disperse the bacteria, then take 1mL of the bacterial suspension on a sterile plate and put it in a 254nm ultraviolet mutagenesis box for 90s, then dilute it 100 times and spread it on bromine containing 20mg / L On cresyl violet plates, cultured anaerobically at 37°C for 3 days. A total of 120 colonies with large colonies, early discoloration time, and larger and brighter discoloration circles were selected as primary strains, and fermented after expanded cultivation to verify yield and stability. Finally, a strain of Clostridium acetobutylicum B3 (Clostridium...

Embodiment 2

[0059]Fermentation medium: glucose 60g / L, ammonium acetate 3g / L, corn steep liquor 2ml / L, sterilized at 121°C for 15min.

[0060] After the seed solution was mixed, it was inserted into a 5L fermenter with a 10% inoculum amount, and the liquid amount was 60%, and it was cultured statically at 37° C. for 80 h.

[0061] In the fermented liquid finally obtained, acetone was 3.9 g / L, ethanol was 0.8 g / L, butanol was 11.9 g / L, and 3-hydroxybutanone was 2.4 g / L.

Embodiment 3

[0063] Fermentation medium: 6% corn flour, gelatinized in boiling water for 60 minutes, sterilized at 121°C for 30 minutes.

[0064] After the seed solution was mixed, it was inserted into a 5L fermenter with a 5% inoculum size, and the liquid volume was 3L, and it was cultured statically at 37° C. for 50 h.

[0065] In the fermentation broth finally obtained, acetone was 4.2g / L, ethanol was 1.1g / L, butanol was 11.8g / L, and 3-hydroxybutanone was 2.1g / L.

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Abstract

The invention provides a Clostridium acetobutylicum and applications thereof. The accession number of the Clostridium acetobutylicum provided by the invention is CGMCC No. 5234. The Clostridium acetobutylicum can be used to simultaneously produce acetone, butanol, ethanol and 3-hydroxybutanone through fermentation, so that economic benefits of butanol fermentation are increased. Coupled regeneration of NAD+ can be achieved by adding metabolism or growth regulators, which increases product yields and, at the same time, can flexibly adjust the yields of coproduction products to adapt to market demand.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to Clostridium acetobutylicum producing 3-hydroxybutanone and an application thereof. Background technique [0002] Acetone (acetone), butanol (butanol) and ethanol (ethanol) are collectively referred to as ABE, which are important raw materials for medicine, pesticides, plastics, rubber and light industry, and are also important chemical solvents. Therefore, the research on ABE is of great importance to the development of modern industry. effect. [0003] Acetone, also known as dimethyl ketone, is a colorless, transparent liquid that is volatile. Acetone is not only an important organic solvent but also a very important chemical raw material. It is used in explosives, plastics, fibers, leather, spray paint and other industries, and can also be used to synthesize ketene, acetic anhydride, iodoform, vinyl rubber, epoxy resin, etc. [0004] Butanol, also known as 1-butanol, is ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/16C12P7/26C12R1/145
Inventor 应汉杰李安柳东陈勇周涛吴菁岚林晓清陈晓春谢婧婧柏建新
Owner NANJING UNIV OF TECH
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