Abstract
A synthetic pathway was engineered in Escherichia coli to produce isopropanol by expressing various combinations of genes from Clostridium acetobutylicum ATCC 824, E. coli K-12 MG1655, Clostridium beijerinckii NRRL B593, and Thermoanaerobacter brockii HTD4. The strain with the combination of C. acetobutylicum thl (acetyl-coenzyme A [CoA] acetyltransferase), E. coli atoAD (acetoacetyl-CoA transferase), C. acetobutylicum adc (acetoacetate decarboxylase), and C. beijerinckii adh (secondary alcohol dehydrogenase) achieved the highest titer. This strain produced 81.6 mM isopropanol in shake flasks with a yield of 43.5% (mol/mol) in the production phase. To our knowledge, this work is the first to produce isopropanol in E. coli, and the titer exceeded that from the native producers.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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2-Propanol / metabolism*
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Acetyl-CoA C-Acetyltransferase / genetics
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Alcohol Oxidoreductases / genetics
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Bacterial Proteins / genetics
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Biosynthetic Pathways*
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Carboxy-Lyases / genetics
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Cloning, Molecular
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Clostridium acetobutylicum / enzymology
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Clostridium acetobutylicum / genetics
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Clostridium beijerinckii / enzymology
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Clostridium beijerinckii / genetics
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Escherichia coli / enzymology
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Escherichia coli / genetics*
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Escherichia coli / metabolism*
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Gene Expression
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Thermoanaerobacter / enzymology
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Thermoanaerobacter / genetics
Substances
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Bacterial Proteins
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Alcohol Oxidoreductases
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isopropanol dehydrogenase (NADP)
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Acetyl-CoA C-Acetyltransferase
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Carboxy-Lyases
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acetoacetate decarboxylase
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2-Propanol