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Literature summary extracted from

  • Bai, Y.; van der Kaaij, R.M.; Leemhuis, H.; Pijning, T.; van Leeuwen, S.S.; Jin, Z.; Dijkhuizen, L.
    Biochemical characterization of the Lactobacillus reuteri glycoside hydrolase family 70 GTFB type of 4,6-alpha-glucanotransferase enzymes that synthesize soluble dietary starch fibers (2015), Appl. Environ. Microbiol., 81, 7223-7232.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.B34 expression in Escherichia coli Limosilactobacillus reuteri

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.B34 additional information removal of the variable N-terminal region of the GTFB enzyme (yielding construct GTFB734-1619) results in increased expression of soluble and active enzyme in Escherichia coli Limosilactobacillus reuteri

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.B34 Cu2+ 1 mM, 49% inhibition of hydrolysis reaction, complete inhibition of transfer reaction Limosilactobacillus reuteri
2.4.1.B34 EDTA 1 mM, 25% inhibition of hydrolysis reaction, transfer reaction increases to 388% Limosilactobacillus reuteri
2.4.1.B34 Fe2+ 1 mM, 50% inhibition of hydrolysis reaction, transfer reaction increases to 175% Limosilactobacillus reuteri
2.4.1.B34 Fe3+ 1 mM, 65% inhibition of hydrolysis reaction, 15% inhibition of transfer reaction Limosilactobacillus reuteri
2.4.1.B34 K+ 1 mM, 14% inhibition of hydrolysis reaction, transfer reaction to 435% of initial value, respectively Limosilactobacillus reuteri
2.4.1.B34 Mg2+ 1 mM, 6% inhibition of hydrolysis reaction, transfer reaction increases to 378% of initial value, respectively Limosilactobacillus reuteri

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.B34 additional information
-
 amylose V Km value for hydrolysis reaction 0.5 g/l, for transfer reaction 0.69 g/l, pH 5.0, 55°C Limosilactobacillus reuteri

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.B34 Ca2+ 1 mM, hydrolysis reaction increases to 111%, transfer reaction to 422% of initial value, respectively Limosilactobacillus reuteri
2.4.1.B34 EDTA 1 mM, 25% inhibition of hydrolysis reaction, transfer reaction increases to 388% Limosilactobacillus reuteri
2.4.1.B34 Fe2+ 1 mM, 50% inhibition of hydrolysis reaction, transfer reaction increases to 175% Limosilactobacillus reuteri
2.4.1.B34 K+ 1 mM, 14% inhibition of hydrolysis reaction, transfer reaction to 435% of initial value, respectively Limosilactobacillus reuteri
2.4.1.B34 Mg2+ 1 mM, 6% inhibition of hydrolysis reaction, transfer reaction increases to 378% of initial value, respectively Limosilactobacillus reuteri
2.4.1.B34 Mn2+ 1 mM, hydrolysis reaction increases to 125%, transfer reaction to 425% of initial value, respectively Limosilactobacillus reuteri
2.4.1.B34 Na+ 1 mM, hydrolysis reaction increases to 118%, transfer reaction to 404% of initial value, respectively Limosilactobacillus reuteri

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.B34 Limosilactobacillus reuteri Q5SBM0
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.B34 amylose V + H2O
-
 Limosilactobacillus reuteri ? both hydrolysis and transferase activities occur. Besides the presence of alpha1->4 linkages, alpha1->6 linkages are newly formed. In the product mixture derived from amylose V, the alpha1->6/alpha1->4 linkage ratio increases up to 90:10. In the product mixture, free glucose, 4-substituted reducing glucose residues [-(1->4)-alpha-D-Glc], and a small amount of reducing-end glucose residues which are 6 substituted are detected ?
2.4.1.B34 maltodextrin + H2O i.e. short-chain alpha1->4-linked maltodextrins Limosilactobacillus reuteri ?
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.B34 55
-
 both hydrolysis and transfer reactions Limosilactobacillus reuteri

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.B34 40
-
 stable for more than 2 h Limosilactobacillus reuteri
2.4.1.B34 55
-
 half-life below 10 min Limosilactobacillus reuteri

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.B34 9.06
-
 amylose V hydrolysis reaction, pH 5.0, 55°C Limosilactobacillus reuteri
2.4.1.B34 36.22
-
 amylose V transfer reaction, pH 5.0, 55°C Limosilactobacillus reuteri

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.B34 5
-
 both hydrolysis and transfer reactions Limosilactobacillus reuteri

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.B34 additional information
-
 amylose V Km/kcat value for hydrolysis reaction 18.5 l per s and g, for transfer reaction 53.0 l/s and g, pH 5.0, 55°C Limosilactobacillus reuteri