Literature summary for 1.4.1.13 extracted from

Stabile, H.; Curti, B.; Vanoni, M.A.
Functional properties of recombinant Azospirillum brasilense glutamate synthase, a complex iron-sulfur flavoprotein (2000), Eur. J. Biochem., 267, 2720-2730.

Search for more literature for 1.4.1.13

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the holoenzyme in Escherichia coli	Azospirillum brasilense

Inhibitors

Inhibitors	Comment	Organism	Structure
2',5'-diphosphoadenylic acid	competitive inhibitor with respect to NADPH	Azospirillum brasilense
2'-phosphoadenosine-5'-diphospho-5'-beta-D-ribose	-	Azospirillum brasilense
2'-phosphoadenylic acid	competitive inhibitor with respect to NADPH	Azospirillum brasilense
3-Aminopyridine adenine dinucleotide phosphate	-	Azospirillum brasilense
acetyl-NADP+	-	Azospirillum brasilense
NADP+	-	Azospirillum brasilense
thio-NADP+	-	Azospirillum brasilense

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Iron	iron-sulfur protein	Azospirillum brasilense
Iron	contains 12.05 mol Fe2+ per mol alphabeta protomer	Azospirillum brasilense

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
200000	-	approximate value, the alphabeta protomeric form prevails at low protein concentration and at high ionic strength, gel filtration	Azospirillum brasilense
800000	-	approximate value, the tetrameric form is found at high protein concentration and low ionic strength, gel filtration	Azospirillum brasilense

Organism

Organism	UniProt	Comment	Textmining
Azospirillum brasilense	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
of the recombinant enzyme from overproducing Escherichia coli cells, using ion exchange chromatography on Q-Sepharose, gel filtration on Sephacryl S-300 and affinity chromatography on 2',5' ADP-Sepharose 4B colum	Azospirillum brasilense

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	-	Azospirillum brasilense
17.9	-	-	Azospirillum brasilense

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-glutamine + 2-oxoglutarate + NADPH + H+	-	Azospirillum brasilense	L-glutamate + NADP+	-	?
additional information	the recombinant enzyme has diaphorase activity, it can oxidize NADPH and transfer electrons to synthetic electron acceptors like iodonitrotetrazolium and ferricyanide	Azospirillum brasilense	?	-	?

Cofactor

Cofactor	Comment	Organism	Structure
flavin	flavoenzyme	Azospirillum brasilense
flavin	contains FAD and FMN	Azospirillum brasilense
flavin	0.83 mol FAD and 0.86 mol FMN per mol alphabeta protomer	Azospirillum brasilense
additional information	the enzyme contains three different iron-sulfur clusters, one 3Fe-4S center on the alpha subunit and two 4Fe-4S clusters of unknown location, 11.7 mol sulfur per mol alphabeta protomer	Azospirillum brasilense
NADPH	-	Azospirillum brasilense

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
additional information	-	additional information	-	Azospirillum brasilense
0.00023	-	thio-NADP+	-	Azospirillum brasilense
0.0024	-	3-Aminopyridine adenine dinucleotide phosphate	-	Azospirillum brasilense
0.0028	-	2'-phosphoadenosine 5'-diphospho-5-beta-D-ribose	-	Azospirillum brasilense
0.0058	-	acetyl-NADP+	-	Azospirillum brasilense
0.0059	-	NADP+	-	Azospirillum brasilense
0.048	-	2',5'-diphosphoadenylic acid	-	Azospirillum brasilense
0.186	-	2'-phosphoadenylic acid	-	Azospirillum brasilense

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Release 2023.1 (January 2023)