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Continuous single vessel butanol synthesis by fermentation

A reactor and butanol technology, applied in fermentation, microbes, biofuels, etc., can solve problems such as increasing production costs

Inactive Publication Date: 2012-05-02
尤金 T 巴特勒三世
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, all of the above methods increase the production cost

Method used

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  • Continuous single vessel butanol synthesis by fermentation
  • Continuous single vessel butanol synthesis by fermentation
  • Continuous single vessel butanol synthesis by fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0069] Embodiment 1. method.

[0070] Species-specific PCR.

[0071] According to the records of Klijn et al. in "Appl Env Microbiology", Volume 61, Issue 8, Page 2919-2924, 1995, specific amplification of the V6 region of the 16S rRNA gene was performed by PCR. Reactions were performed in sterile 0.5 ml tubes containing 50 µL of the following buffers: 10 mM Tris-HCl (pH 8.8), 3.0 mM MgCl 2 , 50mM NaCl, 2.5mM (each) deoxynucleoside triphosphate, 1U of Taq Polymerase and 15 ng (each) of primers P1 5'-GCG GCG TGC CTA ATA CAT GC-3' (SEQ ID NO: 1) and P2 5'- GGG TTG CGC TCG TTG CGG GA-3' (SEQ ID NO: 2). After heating to 95 °C to eliminate protease activity, add 5 µL of template DNA (i.e., DNA extracted from the culture). The DNA was melted at 94°C for 1 min, annealed at 55°C for 1.5 min and extended at 72°C for 2.5 min, and the amplification was completed by 30 cycles.

[0072] The resulting PCR product was diluted 10 times, and then 5 μl of the diluted solution was used a...

Embodiment 2

[0093] Example 2. Purification of butanol from fermentation culture by vacuum distillation

[0094] use C. acetobutylicum or C. beijerinkii One problem with the fermentative production of butanol is that butanol is toxic to cells. The maximum tolerable butanol concentration was about 2% (wt / v) regardless of the sugar concentration.

[0095] A possible solution to this problem was attempted, namely vacuum distillation of the butanol / water azeotrope. The boiling point of butanol is about 118°C, while that of water is 100°C. However, if a mixture of butanol and water is heated at atmospheric pressure, the low boiling azeotrope of butanol and water will evaporate at 93°C. The vapor composition of the azeotrope is about 55.5 wt% butanol and 45.5 wt% water. The solubility of butanol in water is only 7.7% by weight, so the condensate will form two layers. The upper layer is 58 mol% butanol and 42 mol% water: ie, by weight, 80% butanol / 20% water. The lower layer contained 7...

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Abstract

The present invention describes a method for producing butanol by fermentation of carbohydrates using mixed populations of acidogenic-phase cells and solventogenic-phase cells of Clostridium in a solitary vessel. The present system as described does not require intermittent adjustment of pH or venting of headspace gases. The method provides a process for removal of the butanol product which does not irreversibly harm the cells and conditions are described where such cells may resume butanol synthesis in the same solitary vessel. The invention also describes compositions and biologically pure cultures which comprise the Clostridium cells as disclosed.

Description

[0001] Cross References to Related Applications [0002] This application is a continuation-in-part of US Patent 12 / 391,026, filed February 23, 2009, which is now obsolete, the disclosure of which is incorporated herein by reference in its entirety. technical field [0003] The present invention relates generally to the production of butanol by fermentation, and in particular to single vessel butanol production by creating microenvironments to maintain subpopulations of acidogenic and solventogenic phase coincident cells. Background technique [0004] Butanol (n-butanol or n-butyl alcohol) can be produced by the fermentation of carbohydrates that degrade into products such as sugars containing five and six carbon atoms ( such as glucose). Charles Weizmann developed this method during World War I (see, eg, US Patents 1,315,585; 1,329,214; 1,437,697). Briefly, the Weizmann method involves the addition of Fusobacterium acetobutylicum ( Clostridium acetobutylicum ) in the pre...

Claims

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Application Information

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IPC IPC(8): C12P7/16C12R1/145
CPCC12N1/20C12R1/145C12P7/16Y02E50/10C12N1/205C12R2001/145
Inventor 尤金·T·巴特勒三世
Owner 尤金 T 巴特勒三世