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Literature summary 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 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.10 C154S site-directed mutagenesis Saccharomyces cerevisiae
1.5.1.10 C154S/Y99F site-directed mutagenesis Saccharomyces cerevisiae
1.5.1.10 D126A site-directed mutagenesis Saccharomyces cerevisiae
1.5.1.10 D126A/C154S site-directed mutagenesis Saccharomyces cerevisiae
1.5.1.10 D126A/Y99F site-directed mutagenesis Saccharomyces cerevisiae
1.5.1.10 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
1.5.1.10 Y99F site-directed mutagenesis Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.10 Saccharomyces cerevisiae P38999
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.5.1.10 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)

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

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.5.1.10 saccharopine reductase
-
 Saccharomyces cerevisiae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.10 7
-
 formation of L-saccharopin Saccharomyces cerevisiae
1.5.1.10 9
-
 formation of L-glutamate Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.10 NADP+
-
 Saccharomyces cerevisiae
1.5.1.10 NADPH
-
 Saccharomyces cerevisiae

General Information

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