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Journal of Molecular Evolution

, Volume 25, Issue 2, pp 151–158 | Cite as

Regulatory mutations that allow the growth ofEscherichia coli on butanol as carbon source

  • David P. Clark
  • Margot L. Rod
Article

Summary

Starting withadhC mutants ofEscherichia coli in which alcohol dehydrogenase (ADH) and acetaldehyde CoA dehydrogenase (ACDH) are expressed constitutively at high levels, we selected mutants with still higher levels of both enzymes. Selection for growth on ethanol in the presence of inhibitors of ADH gave several mutants that had from 2- to 10-fold increases in the levels of both enzymes. These mutations were found to map far from theadhC locus at around 90 min. SuchadhR mutants were unable to grow on acetate or ethanol in certain media unless supplemented with extramanganese. This growth disability was suppressed by secondary mutations, one of which,aceX, increased sensitivity to several toxic metals and may perhaps derepress Mn transport. When theadhR mutation expressing the highest ADH and ACDH levels was present together withfadR andatoC mutations (allowing efficient catabolism of acetoacetyl-CoA) and with anaceX mutation, the resulting strains became capable of usingn-butanol as sole carbon and energy source. The use of butanol byE. coli illustrates the artificial evolution of a new catabolic pathway, in this case by the selection of four successive regulatory mutations (fadR, adhC, atoC, andadhR) together with the poorly definedaceX mutation. Each stage in the acquisition of this nove pathway confers the ability to use a new growth substrate: decanoic acid (fadR), ethanol (adhC), butyric acid (atoC), and butanol (adhR, when present withaceX).

Key words

Experimental evolution Acetaldehyde dehydrogenase Butanol oxidation Alcohol dehydrogenase 

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Copyright information

© Springer-Verlag New York Inc 1987

Authors and Affiliations

  • David P. Clark
    • 1
  • Margot L. Rod
    • 1
  1. 1.Department of MicrobiologySouthern Illinois UniversityCarbondaleUSA

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