PubAg

Main content area

Biodegradation of dioxins by recombinant Escherichia coli expressing rat CYP1A1 or its mutant

Author:
Shinkyo, Raku, Kamakura, Masaki, Ikushiro, Shin-ichi, Inouye, Kuniyo, Sakaki, Toshiyuki
Source:
Applied microbiology and biotechnology 2006 v.72 no.3 pp. 584-590
ISSN:
0175-7598
Subject:
Escherichia coli, biodegradation, bioremediation, cell culture, humans, metabolism, metabolites, mutants, polychlorinated dibenzodioxins, prokaryotic cells, rats, soil, toxicity, yeasts
Abstract:
Among polychlorinated dibenzo-p-dioxins (PCDDs), 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TetraCDD) is the most toxic one. Recently, we reported that rat CYP1A1 mutant, F240A, expressed in yeast showed metabolic activity toward 2,3,7,8-TetraCDD. In this study, we successfully expressed N-terminal truncated P450s (Δ1A1 and ΔF240A) in Escherichia coli cells. Kinetic analysis using membrane fractions prepared from the recombinant E. coli cells revealed that ΔF240A has enzymatic properties similar to F240A expressed in yeast. The metabolism of PCDDs by recombinant E. coli cells expressing both ΔF240A and human NADPH-P450 reductase was also examined. When 2,3,7-TriCDD was added to the E. coli cell culture at a final concentration of 10 μM, approximately 90% of the 2,3,7-TriCDD was converted into multiple metabolites within 8 h. These results indicate the possible application of prokaryotic cells expressing ΔF240A to the bioremediation of PCDD-contaminated soil.
Agid:
629355