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Molecular and General Genetics MGG

, Volume 205, Issue 3, pp 487–493 | Cite as

The use of suicide substrates to select mutants of Escherichia coli lacking enzymes of alcohol fermentation

  • Philip R. Cunningham
  • David P. Clark
Original Articles

Summary

Mutants of Escherichia coli resistant to chloroethanol or to chloroacetaldehyde were selected. Such mutants were found to lack the fermentative coenzyme A (CoA) linked acetaldehyde dehydrogenase activity. Most also lacked the associated fermentative enzyme alcohol dehydrogenase. Both types of mutants, those lacking acetaldehyde dehydrogenase alone or lacking both enzymes, mapped close to the regulatory adhC gene at 27 min on the E. coli genetic map. The previously described acd mutants which lack acetaldehyde dehydrogenase and which map at 63 min were shown to be pleiotropic, affecting respiration and growth on a variety of substrates. It therefore seems likely that the structural genes for both the acetaldehyde and alcohol dehydrogenases lie in the adhCE operon. This interpretation was confirmed by the isolation of temperature sensitive chloracetaldehyde-resistant mutants, some of which produced thermolabile acetaldehyde dehydrogenase and alcohol dehydrogenase and were also found to map at the adh locus. Reversion analysis indicated that mutants lacking one or both enzymes carried single mutations. The gene order in the adh region was determined by three point crosses to be trp - zch:: Tn10 - adh - galU- bglY - tyrT - chlC.

Key words

Acetaldehyde dehydrogenase Alcohol dehydrogenase Chloracetaldehyde Anaerobic growth Ethanol 

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Philip R. Cunningham
    • 1
  • David P. Clark
    • 1
  1. 1.Department of MicrobiologySouthern Illinois UniversityCarbondale

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