BIOMAT Database Logo BIOMAT Database Logo

Purification and characterization of NADP-dependent glutamate dehydrogenase from the commercial mushroom Agaricus bisporus. 1995

J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Department of Microbiology, Faculty of Science, University of Nijmegen, The Netherlands.

The nicotinamide adenine dinucleotide phosphate (NADP)-dependent glutamate dehydrogenase (NADP-GDH) of Agaricus bisporus, a key enzyme in ammonia assimilation, was purified to apparent electrophoretic homogeneity with 27% recovery of the initial activity. The molecular weight of the native enzyme was 330 kDa. The enzyme is probably a hexamer, composed of identical subunits of 48 kDa. The isoelectric point of the enzyme was found at pH 4.8. The N-terminus appeared to be blocked. The enzyme was specific for NADP(H). The Km-values were 2.1, 3.2, 0.074, 27.0, and 0.117 mM for ammonia, 2-oxoglutarate, NADPH, L-glutamate, and NADP respectively. The pH optima for the amination and deamination reactions were found to be 7.6 and 9.0, respectively. The temperature optimum was 33 degrees C. The effect of several metabolites on the enzyme's activity was tested. Pyruvate, oxaloacetate, ADP, and ATP showed some inhibitory effect. Divalent cations slightly stimulated the aminating reaction. Antibodies raised against the purified enzyme were able to precipitate NADP-GDH activity from a cell-free extract in an anticatalytic immunoprecipitation test. Analysis of a Western blot showed the antibodies to be specific for NADP-GDH.

UI MeSH Term Description Entries
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D002264 Carboxylic Acids Organic compounds containing the carboxy group (-COOH). This group of compounds includes amino acids and fatty acids. Carboxylic acids can be saturated, unsaturated, or aromatic. Carboxylic Acid,Acid, Carboxylic,Acids, Carboxylic
D000227 Adenine Nucleotides Adenine Nucleotide,Adenosine Phosphate,Adenosine Phosphates,Nucleotide, Adenine,Nucleotides, Adenine,Phosphate, Adenosine,Phosphates, Adenosine
D000364 Agaricus A basidiomycetous fungal genus of the family Agaricaceae, order Agaricales, which includes the field mushroom (A. campestris) and the commercial mushroom (A. bisporus).
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody
D018086 Glutamate Dehydrogenase (NADP+) An enzyme that catalyzes the conversion of L-glutamate, H2O, and NADP+ to 2-oxoglutarate, NH3, and NADPH. (From Enzyme Nomenclature, 1992) EC 1.4.1.4. NADP+-Dependent Glutamate Dehydrogenase,NADP-Dependent Glutamate Dehydrogenase,NADP-Specific Glutamate Dehydrogenase,NADPH-Dependent Glutamate Dehydrogenase,Dehydrogenase, NADP+-Dependent Glutamate,Dehydrogenase, NADP-Dependent Glutamate,Dehydrogenase, NADP-Specific Glutamate,Dehydrogenase, NADPH-Dependent Glutamate,Glutamate Dehydrogenase, NADP+-Dependent,Glutamate Dehydrogenase, NADP-Dependent,Glutamate Dehydrogenase, NADP-Specific,Glutamate Dehydrogenase, NADPH-Dependent,NADP Dependent Glutamate Dehydrogenase,NADP Specific Glutamate Dehydrogenase,NADP+ Dependent Glutamate Dehydrogenase,NADPH Dependent Glutamate Dehydrogenase

Related Publications

J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and characterization of trehalose phosphorylase from the commercial mushroom Agaricus bisporus.
September 1998, Biochimica et biophysica acta,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and characterization of an acid phosphatase from the commercial mushroom Agaricus bisporus.
April 2000, Antonie van Leeuwenhoek,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and characterization of a serine proteinase from senescent sporophores of the commercial mushroom Agaricus bisporus.
June 1993, Journal of general microbiology,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and characterization of four isolectins of mushroom (Agaricus bisporus).
March 1985, Biological chemistry Hoppe-Seyler,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and Characterization of β-Glucosidase from Agaricus bisporus (White Button Mushroom).
December 2015, The protein journal,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and characterization of the NADP-dependent glutamate dehydrogenase from Bacillus fastidiosus.
April 1989, Antonie van Leeuwenhoek,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Cloning and characterization of NADP-mannitol dehydrogenase cDNA from the button mushroom, Agaricus bisporus, and its expression in response to NaCl stress.
December 1998, Applied and environmental microbiology,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Mushroom (Agaricus bisporus).
January 2006, Methods in molecular biology (Clifton, N.J.),
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Lignin-Degrading Enzymes of the Commercial Button Mushroom, Agaricus bisporus.
March 1994, Applied and environmental microbiology,
J J Baars, and H J Op den Camp, and A H van Hoek, and C van der Drift, and L J Van Griensven, and J Visser, and G D Vogels
Purification and characterization of RNA polymerase II resistant to alpha-amanitin from the mushroom Agaricus bisporus.
March 1979, Biochemistry,
Copied contents to your clipboard!