Modified microorganisms and methods of making butadiene using same
A technology of butadiene and microorganisms, applied in the field of microorganisms, can solve problems such as expensive catalysts, toxicity, etc.
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example 1
[0239] Example 1 : Modified microorganisms for the production of butadiene.
[0240] Microorganisms such as bacteria can be genetically modified to produce butadiene from fermentable carbon sources including, for example, glucose.
[0241] In an exemplary method, a microorganism can be genetically engineered by any method known in the art to include: i.) one or more enzymes in the coding pathway that catalyze the conversion of a fermentable carbon source to acetyl-CoA and propionyl-CoA polynucleotides, and / or polynucleotides for enzymes in one or more coding pathways that catalyze the conversion of acetyl-CoA and propionyl-CoA to butadiene; ii.) in one or more coding pathways that catalyze the conversion of fermentable carbon sources to croton A polynucleotide for an enzyme that converts crotonyl-CoA, and / or a polynucleotide for an enzyme in one or more coding pathways that catalyzes the conversion of crotonyl-CoA to butadiene; or iii.) a polynucleotide that catalyzes a ferm...
example 2
[0243] Example 2 : Fermentation of carbon sources by genetically modified microorganisms to produce butadiene.
[0244] A genetically modified microorganism produced as in Example 1 above can be used to ferment a carbon source to produce butadiene.
[0245] In an exemplary method, a fermentable carbon source (e.g., 9 g / L glucose, 1 g / L KH 2 PO 4 , 2g / L (NH 4 ) 2 HPO 4 , 5mg / L FeSO 4 ·7H 2 O, 10mg / L MgSO 4 ·7H 2 O, 2.5mg / L MnSO 4 ·H 2 O, 10mg / L CaCl 2 ·6H 2 O, 10mg / L CoCl 2 ·6H 2 0, and 10 g / L yeast extract) pre-sterilized medium was fed into the bioreactor.
[0246] During fermentation, anaerobic conditions are maintained by, for example, sparging nitrogen into the medium. A temperature suitable for fermentation (eg, about 30°C) is maintained using any method known in the art. An approximate physiological pH (eg, about 6.5) is maintained by, for example, automatic addition of sodium hydroxide. The bioreactor is stirred eg at about 50 rpm. Let the fermentati...
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