Literature summary for 1.5.1.10 extracted from

Vashishtha, A.K.; West, A.H.; Cook, P.F.
Probing the chemical mechanism of saccharopine reductase from Saccharomyces cerevisiae using site-directed mutagenesis (2015), Arch. Biochem. Biophys., 584, 98-106 .

Search for more literature for 1.5.1.10

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain LYS 9	Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment)	Organism
analysis of the apoenzyme crystal structure determined at 1.7 A resolution	Saccharomyces cerevisiae

Protein Variants

Protein Variants	Comment	Organism
C154S	site-directed mutagenesis	Saccharomyces cerevisiae
C154S/Y99F	site-directed mutagenesis	Saccharomyces cerevisiae
D126A	site-directed mutagenesis	Saccharomyces cerevisiae
D126A/C154S	site-directed mutagenesis	Saccharomyces cerevisiae
D126A/Y99F	site-directed mutagenesis	Saccharomyces cerevisiae
additional information	kinetic parameters of the mutants in the reaction direction of glutamate formation exhibit modest decreases. The pH-rate profiles obtained with all mutant enzymes decrease at low and high pH, suggesting acid and base catalytic groups are still present in all enzymes. Solvent kinetic deuterium isotope effects are all larger than those observed for wild-type enzyme	Saccharomyces cerevisiae
Y99F	site-directed mutagenesis	Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
N6-(L-1,3-dicarboxypropyl)-L-lysine + NADP+ + H2O	Saccharomyces cerevisiae	-	L-glutamate + (S)-2-amino-6-oxohexanoate + NADPH + H+	-	r

Organism

Organism	UniProt	Comment	Textmining
Saccharomyces cerevisiae	P38999	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
N6-(L-1,3-dicarboxypropyl)-L-lysine + NADP+ + H2O = L-glutamate + (S)-2-amino-6-oxohexanoate + NADPH + H+	ordered kinetic mechanism, the reduced dinucleotide substrate binds to enzyme first followed by L-alpha-aminoadipate-delta-semialdehyde, which adds in rapid equilibrium prior to L-glutamate, saccharopine is released prior to NADP+, primary deuterium kinetic isotope effects and solvent deuterium kinetic isotope effects, overview. A conformational change to open the site and release products (in the direction of saccharopine formation) is the rate limiting step. Two groups are involved in the acid-base chemistry of the reaction. An enzyme group with a pKa of 5.6 accepts a proton from the alpha-amine of glutamate to generate the neutral amine that can act as a nucleophile. The alpha-amine of glutamate attacks the carbonyl of the semialdehyde to generate the carbinolamine, which is protonated by a second enzyme group with a pKa of about 7.8-8.0. Ionizable residues that might play a role in the acid-base mechanism of the enzyme are D126, C154 and/or Y99	Saccharomyces cerevisiae	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
N6-(L-1,3-dicarboxypropyl)-L-lysine + NADP+ + H2O	-	Saccharomyces cerevisiae	L-glutamate + (S)-2-amino-6-oxohexanoate + NADPH + H+	-	r

Subunits

Subunits	Comment	Organism
More	comparison of enzyme structures from Saccharomyces cerevisiae and Magnoporthe grisea	Saccharomyces cerevisiae

Synonyms

Synonyms	Comment	Organism
saccharopine reductase	-	Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	formation of L-saccharopin	Saccharomyces cerevisiae
9	-	formation of L-glutamate	Saccharomyces cerevisiae

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	-	Saccharomyces cerevisiae
NADPH	-	Saccharomyces cerevisiae

General Information

General Information	Comment	Organism
physiological function	saccharopine reductase catalyzes the reductive amination of L-alpha-aminoadipate-delta-semialdehyde with L-glutamate to give saccharopine	Saccharomyces cerevisiae

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Release 2023.1 (January 2023)