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Literature summary for 1.5.1.7 extracted from

  • Kumar, V.P.; Thomas, L.M.; Bobyk, K.D.; Andi, B.; Cook, P.F.; West, A.H.
    Evidence in support of lysine 77 and histidine 96 as acid-base catalytic residues in saccharopine dehydrogenase from Saccharomyces cerevisiae (2012), Biochemistry, 51, 857-866.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.1.7

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 (DE3)-RIL cells Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme C205S, hanging drop vapor diffusion method, using 100 mM Tris (pH 7.0), 30% (w/v) PEG-MME 2000 at 4°C Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
H96Q the mutation results in 100 and more than 1000fold increases in Km values for L-lysine and 2-oxoglutarate, respectively Saccharomyces cerevisiae
K77M the mutation results in 28 and 90fold increases in Km values for L-lysine and 2-oxoglutarate, respectively Saccharomyces cerevisiae
K77M/H96Q the mutations result in 300 and 80fold increases in Km values for L-lysine and 2-oxoglutarate, respectively Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
 2-oxoglutarate wild type enzyme, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
0.89
-
 L-lysine wild type enzyme, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
9
-
 2-oxoglutarate mutant enzyme H96Q, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
10
-
 2-oxoglutarate mutant enzyme K77M, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
25
-
 L-lysine mutant enzyme K77M, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
96
-
 L-lysine mutant enzyme K77M/H96Q, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
267
-
 L-lysine mutant enzyme H96Q, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae
267
-
 2-oxoglutarate mutant enzyme K77M/H96Q, in 100 mM HEPES, pH 7.0, at 25°C Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-lysine + 2-oxoglutarate + NADH Saccharomyces cerevisiae
-
 N6-(L-1,3-dicarboxypropyl)-L-lysine + NAD+ + H2O
-
 r

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P38998
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA affinity column chromatography Saccharomyces cerevisiae

Storage Stability

Storage Stability Organism
4°C, mutant enzymes in 100 mM HEPES, 300 mM KCl, and 300 mM imidazole at pH 8.0, several months, no loss of activity Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-lysine + 2-oxoglutarate + NADH
-
 Saccharomyces cerevisiae N6-(L-1,3-dicarboxypropyl)-L-lysine + NAD+ + H2O
-
 r

Synonyms

Synonyms Comment Organism
N6-(glutaryl-2)-L-lysine: NAD oxidoreductase
-
 Saccharomyces cerevisiae
SDH
-
 Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Saccharomyces cerevisiae