TY  - JOUR
AU  - Piotrowski, M.
AU  - Janowitz, T.
AU  - Kneifel, H.
TI  - Plant C-N-Hydrolases and the identification of a plant N-Carbamoylputrescine amidohydrolase involved in polyamine biosynthesis
JO  - The journal of biological chemistry
VL  - 278
SN  - 0021-9258
CY  - Bethesda, Md.
PB  - Soc.
M1  - PreJuSER-23567
SP  - 1708 - 1712
PY  - 2003
N1  - Record converted from VDB: 12.11.2012
AB  - A nitrilase-like protein from Arabidopsis thaliana (NLP1) was expressed in Escherichia coli as a His(6)-tagged protein and purified to apparent homogeneity by Ni(2+)-chelate affinity chromatography. The purified enzyme showed N-carbamoylputrescine amidohydrolase activity, an enzyme involved in the biosynthesis of polyamines in plants and bacteria. N-carbamoylputrescine amidohydrolase activity was confirmed by identification of two of the three occurring products, namely putrescine and ammonia. In contrast, no enzymatic activity could be detected when applying various compounds including nitriles, amines, and amides as well as other N-carbamoyl compounds, indicating the specificity of the enzyme for N-carbamoylputrescine. Like the homologous beta-alanine synthases, NLP1 showed positive cooperativity toward its substrate. The native enzyme had a molecular mass of 279 kDa as shown by blue-native polyacrylamide gel electrophoresis, indicating a complex of eight monomers. Expression of the NLP1 gene was found in all organs investigated, but it was not induced upon osmotic stress, which is known to induce biosynthesis of putrescine. This is the first report of cloning and expression of a plant N-carbamoylputrescine amidohydrolase and the first time that N-carbamoylputrescine amidohydrolase activity of a recombinant protein could be shown in vitro. NLP1 is one of the two missing links in the arginine decarboxylase pathway of putrescine biosynthesis in higher plants.
KW  - Arabidopsis: enzymology
KW  - Biogenic Polyamines: biosynthesis
KW  - Blotting, Northern
KW  - Electrophoresis, Polyacrylamide Gel
KW  - Mass Spectrometry
KW  - Phylogeny
KW  - Ureohydrolases: chemistry
KW  - Ureohydrolases: genetics
KW  - Ureohydrolases: metabolism
KW  - Biogenic Polyamines (NLM Chemicals)
KW  - N-carbamoylputrescine hydrolase (NLM Chemicals)
KW  - Ureohydrolases (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:12435743
UR  - <Go to ISI:>//WOS:000180462200046
DO  - DOI:10.1074/jbc.M205699200
UR  - https://juser.fz-juelich.de/record/23567
ER  -