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Literature summary extracted from

  • Cook, W.J.; Senkovich, O.; Hernandez, A.; Speed, H.; Chattopadhyay, D.
    Biochemical and structural characterization of Cryptosporidium parvum lactate dehydrogenase (2015), Int. J. Biol. Macromol., 74, 608-619.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.27 analysis CpLDH with APAD+ may be useful as a diagnostic tool for detection of protozoan parasite Cryptosporidium Cryptosporidium parvum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.27 gene LDH1, recombinant expression of the enzyme in Escherichia coli strain Rosetta (DE3)pLysS, molecular replacement and modelling using the structure of PfLDH, PDB ID 1T2D Cryptosporidium parvum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.27 purified recombinant enzyme as apo-enzyme and in four ternary complexes, The ternary complexes capture the enzyme bound to NAD/NADH or its 3-acetylpyridine analogue in the cofactor binding pocket, while the substrate binding site is occupied by one of the following ligands: lactate, pyruvate or oxamate. X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution Cryptosporidium parvum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.27 gossypol
-
 Cryptosporidium parvum
1.1.1.27 additional information CpLDH does not display any measurable inhibition to pyruvate concentrations up to at least 20 mM in contrast to many other LDHs Cryptosporidium parvum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.27 additional information
-
 additional information Michaelis-Menten kinetics Cryptosporidium parvum
1.1.1.27 0.0018
-
 (S)-lactate pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.0085
-
 APAD+ pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.0166
-
 APADH pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.0169
-
 NADH pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.0303
-
 NAD+ pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.1973
-
 pyruvate pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.27 (S)-lactate + NAD+ Cryptosporidium parvum
-
 pyruvate + NADH + H+
-
 r
1.1.1.27 pyruvate + NADH + H+ Cryptosporidium parvum
-
 (S)-lactate + NAD+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.27 Cryptosporidium parvum Q9GT92
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.27 recombinant enzyme from Escherichia coli strain Rosetta (DE3)pLysS by ammonium sulfate fractionation, gel filtration, anion exchange chromatography, and ultrafiltration Cryptosporidium parvum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.27 (S)-lactate + APAD+
-
 Cryptosporidium parvum pyruvate + APADH + H+
-
 r
1.1.1.27 (S)-lactate + NAD+
-
 Cryptosporidium parvum pyruvate + NADH + H+
-
 r
1.1.1.27 pyruvate + APADH + H+
-
 Cryptosporidium parvum (S)-lactate + APAD+
-
 r
1.1.1.27 pyruvate + NADH + H+
-
 Cryptosporidium parvum (S)-lactate + NAD+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.27 More the ternary complexes capture the enzyme bound to NAD/NADH or its 3-acetylpyridine analog in the cofactor binding pocket, while the substrate binding site is occupied by one of the following ligands: lactate, pyruvate or oxamate Cryptosporidium parvum

Synonyms

EC Number Synonyms Comment Organism
1.1.1.27 lactate dehydrogenase
-
 Cryptosporidium parvum
1.1.1.27 LDH1
-
 Cryptosporidium parvum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.27 25
-
 assay at Cryptosporidium parvum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.27 0.000045
-
 (S)-lactate pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.000055
-
 NAD+ pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.000248
-
 APAD+ pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.00208
-
 NADH pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.00209
-
 pyruvate pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.00386
-
 APADH pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.27 5 5.5 reduction of pyruvate Cryptosporidium parvum
1.1.1.27 8.5 9.5 oxidation of lactate Cryptosporidium parvum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.27 4 8 over 40% of maximal activity for reduction of pyruvate within this range Cryptosporidium parvum
1.1.1.27 8 9.5 20% of maximal activity for oxidation of lactate at pH 8.0, 90% at pH 8.5, and maximal activity at pH 8.5-9.5, inactive at pH 7.5 Cryptosporidium parvum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.27 3-acetyl-pyridine adenine dinucleotide i.e. APAD+, 3-acetylpyridine adenine dinucleotide, analogue of NAD+ Cryptosporidium parvum
1.1.1.27 additional information Cryptosporidium parvum lactate dehydrogenase prefers the acetylpyridine analogue of NADH as a cofactor. NADH/APADH and substrate binding site structures, overview. replacement of the conserved serine residue by Met163 in CpLDH allows a better packing environment for the acetyl of APAD+ Cryptosporidium parvum
1.1.1.27 NAD+
-
 Cryptosporidium parvum
1.1.1.27 NADH
-
 Cryptosporidium parvum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.27 0.0116
-
 gossypol pH 5.5, 25°C, recombinant enzyme, with NADH and pyruvate Cryptosporidium parvum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.27 0.000033
-
 (S)-lactate pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.00183
-
 NAD+ pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.0105
-
 pyruvate pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.0292
-
 APAD+ pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.1235
-
 NADH pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum
1.1.1.27 0.2326
-
 APADH pH 5.5, 25°C, recombinant enzyme Cryptosporidium parvum