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Literature summary extracted from

  • Hsu, L.J.; Hsu, N.S.; Wang, Y.L.; Wu, C.J.; Li, T.L.
    Structural and biochemical interrogation on transketolase from Pichia stipitis for new functionality (2016), Protein Eng. Des. Sel., 29, 513-522 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.2.1.1 expressed in Escherichia coli BL21(DE3) cells Scheffersomyces stipitis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.2.1.1 wild type and mutants H261F, H66C/261C, R356L and R525L enzymes in complex with various physiological ligands, hanging drop vapor diffusion method, using 0.1 M MES at pH 6.5, 0.1M NaCl, 30% (w/V) PEG 400 Scheffersomyces stipitis

Protein Variants

EC Number Protein Variants Comment Organism
2.2.1.1 H261F inactive Scheffersomyces stipitis
2.2.1.1 H27A in the reaction with sedoheptulose 7-phosphate (backward) the mutant exhibits weaker activity relative to the wild type enzyme Scheffersomyces stipitis
2.2.1.1 R356L in the reaction with D-xylulose 5-phosphate (forward) the mutant exhibits weaker activity relative to the wild type enzyme Scheffersomyces stipitis
2.2.1.1 R525L in the reaction with D-xylulose 5-phosphate (forward) the mutant exhibits weaker activity relative to the wild type enzyme Scheffersomyces stipitis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.2.1.1 0.029
-
 D-fructose 6-phosphate wild type enzyme, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.031
-
 sedoheptulose 7-phosphate wild type enzyme, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.145
-
 D-xylulose 5-phosphate wild type enzyme, with D-erythrose 4-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.183
-
 D-xylulose 5-phosphate wild type enzyme, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.232
-
 D-xylulose 5-phosphate mutant enzyme H27A, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.294
-
 sedoheptulose 7-phosphate mutant enzyme H27A, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.517
-
 D-xylulose 5-phosphate mutant enzyme R356L, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.68
-
 D-xylulose 5-phosphate mutant enzyme R525L, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.2.1.1 D-ribose 5-phosphate + D-xylulose 5-phosphate Scheffersomyces stipitis
-
 sedoheptulose 7-phosphate + D-glyceraldehyde 3-phosphate
-
 r
2.2.1.1 sedoheptulose 7-phosphate + D-glyceraldehyde 3-phosphate Scheffersomyces stipitis
-
 D-ribose 5-phosphate + D-xylulose 5-phosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.2.1.1 Scheffersomyces stipitis P34736
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.2.1.1 Ni2+-NTA agarose resin column chromatography and Superdex S-200 gel filtration Scheffersomyces stipitis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.2.1.1 D-erythrose 4-phosphate + D-xylulose 5-phosphate
-
 Scheffersomyces stipitis D-fructose 6-phosphate + D-glyceraldehyde 3-phosphate
-
 r
2.2.1.1 D-fructose 6-phosphate + D-glyceraldehyde 3-phosphate
-
 Scheffersomyces stipitis D-erythrose 4-phosphate + D-xylulose 5-phosphate
-
 r
2.2.1.1 D-ribose 5-phosphate + D-xylulose 5-phosphate
-
 Scheffersomyces stipitis sedoheptulose 7-phosphate + D-glyceraldehyde 3-phosphate
-
 r
2.2.1.1 sedoheptulose 7-phosphate + D-glyceraldehyde 3-phosphate
-
 Scheffersomyces stipitis D-ribose 5-phosphate + D-xylulose 5-phosphate
-
 r

Subunits

EC Number Subunits Comment Organism
2.2.1.1 homodimer 2 * 74969, calculated from amino acid sequence Scheffersomyces stipitis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.2.1.1 0.35
-
 D-xylulose 5-phosphate mutant enzyme R356L, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.53
-
 D-xylulose 5-phosphate mutant enzyme R525L, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 18.02
-
 D-xylulose 5-phosphate wild type enzyme, with D-erythrose 4-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 18.56
-
 D-xylulose 5-phosphate mutant enzyme H27A, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 20.58
-
 sedoheptulose 7-phosphate mutant enzyme H27A, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 25.31
-
 D-xylulose 5-phosphate wild type enzyme, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 26.87
-
 D-fructose 6-phosphate wild type enzyme, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 28.642
-
 sedoheptulose 7-phosphate wild type enzyme, at pH 8.0 and 25°C Scheffersomyces stipitis

Cofactor

EC Number Cofactor Comment Organism Structure
2.2.1.1 thiamine diphosphate
-
 Scheffersomyces stipitis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.2.1.1 0.66
-
 D-xylulose 5-phosphate mutant enzyme R356L, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 0.78
-
 D-xylulose 5-phosphate mutant enzyme R525L, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 70
-
 sedoheptulose 7-phosphate mutant enzyme H27A, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 80
-
 D-xylulose 5-phosphate mutant enzyme H27A, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 120
-
 D-xylulose 5-phosphate wild type enzyme, with D-erythrose 4-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 140
-
 D-xylulose 5-phosphate wild type enzyme, with D-ribose 5-phosphate as cosubstrate, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 910
-
 sedoheptulose 7-phosphate wild type enzyme, at pH 8.0 and 25°C Scheffersomyces stipitis
2.2.1.1 920
-
 D-fructose 6-phosphate wild type enzyme, at pH 8.0 and 25°C Scheffersomyces stipitis