Purification, properties, and sequence of glycerol trinitrate reductase from Agrobacterium radiobacter

Glycerol trinitrate (GTN) reductase, which enables Agrobacterium radiobacter to utilize GTN and related explosives as sources of nitrogen for growth, was purified and characterized, and its gene was cloned and sequenced. The enzyme was a 39-kDa monomeric protein which catalyzed the NADH-dependent re...

Full description

Saved in:
Bibliographic Details
Published in:Journal of Bacteriology 1997-12, Vol.179 (24), p.7796-7802
Main Authors: Snape, J.R, Walkley, N.A, Morby, A.P, Nicklin, S, White, G.F
Format: Article
Language:English
Subjects:
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
AGROBACTERIUM RADIOBACTER
Amino Acid Sequence
AMINO ACID SEQUENCES
Base Sequence
CHEMICAL COMPOSITION
Cloning, Molecular
COMPOSICION QUIMICA
COMPOSITION CHIMIQUE
DNA Primers
ENZYMIC ACTIVITY
ESTER
ESTERES
ESTERS
EXPLOSIF
EXPLOSIVES
EXPLOSIVOS
GENANK/Y13942
MOLECULAR SEQUENCE DATA
MOLECULAR WEIGHT
NERA GENE
NITRATES (ESTERS)
NUCLEOTIDE SEQUENCE
OPEN READING FRAMES
OXIDOREDUCTASES
Oxidoreductases - genetics
Oxidoreductases - isolation & purification
OXIDORREDUCTASAS
OXYDOREDUCTASE
PESO MOLECULAR
POIDS MOLECULAIRE
Polymerase Chain Reaction
PROMOTERS
PURIFICACION
PURIFICATION
Rhizobium - enzymology
SECUENCIA NUCLEOTIDICA
Sequence Analysis, DNA
Sequence Homology, Amino Acid
SEQUENCE NUCLEOTIDIQUE
Substrate Specificity
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Glycerol trinitrate (GTN) reductase, which enables Agrobacterium radiobacter to utilize GTN and related explosives as sources of nitrogen for growth, was purified and characterized, and its gene was cloned and sequenced. The enzyme was a 39-kDa monomeric protein which catalyzed the NADH-dependent reductive scission of GTN (Km = 23 micromolar) to glycerol dinitrates (mainly the 1,3-isomer) with a pH optimum of 6.5, a temperature optimum of 35 degrees C, and no dependence on metal ions for activity. It was also active on pentaerythritol tetranitrate (PETN), on isosorbide dinitrate, and, very weakly, on ethyleneglycol dinitrate, but it was inactive on isopropyl nitrate, hexahydro-1,3,5-trinitro-1,3,5-triazine, 2,4,6-trinitrotoluene, ammonium ions, nitrate, or nitrite. The amino acid sequence deduced from the DNA sequence was homologous (42 to 51% identity and 61 to 69% similarity) to those of PETN reductase from Enterobacter cloacae, N-ethylmaleimide reductase from Escherichia coli, morphinone reductase from Pseudomonas putida, and old yellow enzyme from Saccharomyces cerevisiae, placing the GTN reductase in the alpha/beta barrel flavoprotein group of proteins. GTN reductase and PETN reductase were very similar in many respects except in their distinct preferences for NADH and NADPH cofactors, respectively
ISSN:0021-9193
1098-5530
1067-8832
DOI:10.1128/jb.179.24.7796-7802.1997