Metabolic activation of aromatic amine mutagens by simultaneous expression of human cytochrome P450 1A2, NADPH-cytochrome P450 reductase, and N-acetyltransferase in Escherichia coli

Chem Res Toxicol. 1998 Jan;11(1):70-4. doi: 10.1021/tx970171q.

Abstract

We describe the construction of a new strain of Escherichia coli designed to bioactivate aromatic amines and to detect their mutagenicity with high sensitivity. Strain DJ4309 bears two plasmids, a pACYC184-derived plasmid which expresses Salmonella typhimurium acetyl CoA:arylamine N-acetyltransferase (NAT) and a pBR322-derived plasmid which expresses human cytochrome P450 1A2 and NADPH-cytochrome P450 reductase. The combined actions of these enzymes convert aromatic amines into reactive, mutagenic N-acetoxy esters. The strain also carries a mutated copy of the lacZ gene (on an F' factor) which reverts to the wild-type gene by a -(GpC) frameshift mutation. Strain DJ4309 expresses high levels of NAT and cytochrome P450 1A2 and is very sensitive to mutagenesis induced by representative aromatic amines. Mutagenicity of 2-aminoanthracene in strain DJ4309 is higher than can be obtained by rat liver homogenate 9000g supernatant (S9) activation in the parent strain lacking the P450 expression vector. Strain DJ4309 provides a useful system for detecting mutagenic aromatic amines and for studying their metabolism by human P450 1A2.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Arylamine N-Acetyltransferase / metabolism*
  • Biotransformation
  • Cytochrome P-450 CYP1A2 / metabolism*
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Humans
  • Hydrocarbons, Aromatic / metabolism*
  • Mutagens / metabolism*
  • NADH, NADPH Oxidoreductases / metabolism*
  • NADPH-Ferrihemoprotein Reductase
  • Plasmids

Substances

  • Hydrocarbons, Aromatic
  • Mutagens
  • Cytochrome P-450 CYP1A2
  • NADH, NADPH Oxidoreductases
  • NADPH-Ferrihemoprotein Reductase
  • Arylamine N-Acetyltransferase