Characterization of Escherichia coli nitroreductase NfsB in the metabolism of nitrobenzodiazepines

Shiuan Woei LinWu, Cheng Jie Syu, Yu Lian Chen, Andrew H.J. Wang, Fu Chuo Peng

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

Abstract

Nitrobenzodiazepine (NBDZ) is a sedative-hypnotic drug used in the treatment of anxiety and sleep problems. Overdose of NBDZ may cause severe neurological effects, especially for people in drug abuse or addiction. In the present study, we investigated NBDZ nitroreduction in rat enteric contents and characterized the role of enterobacterial nitroreductase in the reductive pathway. Nitroreduction of flunitrazepam (FZ) was studied in the microsomal membrane fractions of rat liver, jejunum and jejunal microflora using HPLC analysis. In the jejunal microflora, FZ was demonstrated to be significantly reduced to its amino derivative under anaerobic condition. Escherichia coli type I nitroreductase NfsB (EC 1.5.1.34) was found in rat jejunal microflora via PCR technique and Western blotting. The participation of NfsB in FZ nitroreduction was demonstrated from inhibition studies. Kinetic study of the purified recombinant NfsB indicated that nitroreduction of FZ, nitrazepam (NZ) and clonazepam (CZ) are mediated by NfsB, where CZ shows lower kcat/KM ratio than that of the other two. Finally, two other nitroreductases E. cloacae NR (EC 1.6.99.7) and S. typhimurium Cnr were also found to be responsible for FZ nitroreduction. These results provide that the reduction of NBDZ in normal flora is catalyzed by type I nitroreductase NfsB.

Original languageEnglish
Pages (from-to)96-103
Number of pages8
JournalBiochemical Pharmacology
Volume78
Issue number1
DOIs
Publication statusPublished - Jul 1 2009
Externally publishedYes

Keywords

  • Escherichia coli nitroreductase NfsB
  • Flunitrazepam
  • NADPH-cytochrome P450 reductase
  • Nitrobenzodiazepine
  • Rat jejunal microflora

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

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