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

doi:10.1016/j.bcp.2009.03.019

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 journal › Article › peer-review

21 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 k_cat/K_M 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 language	English
Pages (from-to)	96-103
Number of pages	8
Journal	Biochemical Pharmacology
Volume	78
Issue number	1
DOIs	https://doi.org/10.1016/j.bcp.2009.03.019
Publication status	Published - Jul 1 2009
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

Biochemistry
Pharmacology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bcp.2009.03.019

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@article{9466030823fb4d3bb2450069828b3f39,

title = "Characterization of Escherichia coli nitroreductase NfsB in the metabolism of nitrobenzodiazepines",

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.",

keywords = "Escherichia coli nitroreductase NfsB, Flunitrazepam, NADPH-cytochrome P450 reductase, Nitrobenzodiazepine, Rat jejunal microflora",

author = "LinWu, {Shiuan Woei} and Syu, {Cheng Jie} and Chen, {Yu Lian} and Wang, {Andrew H.J.} and Peng, {Fu Chuo}",

note = "Funding Information: This work was supported by grant number NSC 95-2320-B-002-100 from the National Science Council.",

year = "2009",

month = jul,

day = "1",

doi = "10.1016/j.bcp.2009.03.019",

language = "English",

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T1 - Characterization of Escherichia coli nitroreductase NfsB in the metabolism of nitrobenzodiazepines

AU - LinWu, Shiuan Woei

AU - Syu, Cheng Jie

AU - Chen, Yu Lian

AU - Wang, Andrew H.J.

AU - Peng, Fu Chuo

N1 - Funding Information: This work was supported by grant number NSC 95-2320-B-002-100 from the National Science Council.

PY - 2009/7/1

Y1 - 2009/7/1

N2 - 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.

AB - 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.

KW - Escherichia coli nitroreductase NfsB

KW - Flunitrazepam

KW - NADPH-cytochrome P450 reductase

KW - Nitrobenzodiazepine

KW - Rat jejunal microflora

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M3 - Article

C2 - 19447228

AN - SCOPUS:67349129059

SN - 0006-2952

VL - 78

SP - 96

EP - 103

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

IS - 1

ER -

Characterization of Escherichia coli nitroreductase NfsB in the metabolism of nitrobenzodiazepines

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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