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Literature summary extracted from

  • Sikowitz, M.D.; Shome, B.; Zhang, Y.; Begley, T.P.; Ealick, S.E.
    Structure of a Clostridium botulinum C143S thiaminase I/thiamin complex reveals active site architecture (2013), Biochemistry, 52, 7830-7839.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.2 inactive C143S mutant with bound thiamine at 2.2 A resolution. Each monomer contains one thiamine molecule in a V-shaped cleft between the two domains. Tyrosine residues, Tyr46, Tyr48, Tyr80, Tyr252, Tyr269, and Tyr300, form an outer collar of the cleft with most of the active site residues located at the bottom of the cleft. The catalytic cysteine residue Cys143 is located on beta6 Clostridium botulinum

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.2 C143S inactive Clostridium botulinum
2.5.1.2 D302N about 20fold decrease in kcat/Km value Clostridium botulinum
2.5.1.2 D94N about 10000fold decrease in kcat/Km value Clostridium botulinum
2.5.1.2 E271Q about 3000fold decrease in kcat/Km value Clostridium botulinum
2.5.1.2 Y46F about 2fold decrease in kcat/Km value Clostridium botulinum
2.5.1.2 Y80F about 1000fold decrease in kcat/Km value Clostridium botulinum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.2 0.03
-
 thiamine mutant D302N, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 0.075
-
 thiamine mutant E271Q, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 0.25
-
 thiamine mutant Y46F, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 0.45
-
 thiamine wild-type, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 2.1
-
 thiamine mutant Y80F, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 3.5
-
 thiamine mutant D94N, pH 8.0, 37°C Clostridium botulinum

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.2 Clostridium botulinum
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.2 thiamine + 2-mercaptoethanol
-
 Clostridium botulinum ?
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.2 0.01
-
 thiamine mutant E271Q, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 0.23
-
 thiamine mutant D94N, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 1.1
-
 thiamine mutant Y80F, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 8.1
-
 thiamine mutant D302N, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 60.9
-
 thiamine mutant Y46F, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 231
-
 thiamine wild-type, pH 8.0, 37°C Clostridium botulinum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.2 0.07
-
 thiamine mutant D94N, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 0.15
-
 thiamine mutant E271Q, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 0.5
-
 thiamine mutant Y80F, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 26.4
-
 thiamine mutant D302N, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 252
-
 thiamine mutant Y46F, pH 8.0, 37°C Clostridium botulinum
2.5.1.2 510
-
 thiamine wild-type, pH 8.0, 37°C Clostridium botulinum