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Literature summary for 2.4.1.B34 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
Search for more literature for 2.4.1.B34

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Limosilactobacillus reuteri

Protein Variants

Protein Variants Comment Organism
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

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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

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

Organism

Organism UniProt Comment Textmining
Limosilactobacillus reuteri Q5SBM0
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 ?
maltodextrin + H2O i.e. short-chain alpha1->4-linked maltodextrins Limosilactobacillus reuteri ?
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
 both hydrolysis and transfer reactions Limosilactobacillus reuteri

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
 both hydrolysis and transfer reactions Limosilactobacillus reuteri

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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