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Literature summary extracted from

  • Te Poele, E.; Corwin, S.; Hamaker, B.R.; Lamothe, L.; Vafeiadi, C.; Dijkhuizen, L.
    Development of slowly digestible starch derived alpha-glucans with 4,6-alpha-glucanotransferase and branching sucrase enzymes (2020), J. Agric. Food Chem., 68, 6664-6671 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.B34 gene gtfB, recombinant expression of C-terminally His-tagged enzyme mutant GtfB-DELTAN-DELTAV in Escherichia coli strain BL21(DE3) Limosilactobacillus reuteri
2.4.1.362 gene gtfB, recombinant expression of C-terminally His-tagged enzyme mutant GtfZ-CD2 in Escherichia coli strain BL21(DE3) Apilactobacillus kunkeei

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.B34 additional information construction of the GtfB-DELTAN-DELTAV mutant, a truncated variant (amino acids 761 to 1619) of GtfB of Lactobacillus reuteri strain 121 lacking both the N-terminal variable domain and domain V Limosilactobacillus reuteri
2.4.1.362 additional information construction of the GtfZ-CD2 mutant, a truncated variant comprising the catalytic domain CD2 (amino acids 121-2264) of the GtfZ protein of Lactobacillus kunkeei strain DSM 12361 Apilactobacillus kunkeei

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.B34 Ca2+ required Limosilactobacillus reuteri

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.362 sucrose + (1->6)-alpha-D-glucan Apilactobacillus kunkeei
-
 D-fructose + (1->6)-alpha-D-glucan containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 sucrose + (1->6)-alpha-D-glucan Apilactobacillus kunkeei DSM 12361
-
 D-fructose + (1->6)-alpha-D-glucan containing a (1->3)-alpha-D-glucose branch
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.B34 Limosilactobacillus reuteri A0A0U5F702
-
-
2.4.1.B34 Limosilactobacillus reuteri 121 A0A0U5F702
-
-
2.4.1.362 Apilactobacillus kunkeei
-
-
-
2.4.1.362 Apilactobacillus kunkeei DSM 12361
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.B34 recombinant C-terminally His-tagged enzyme mutant GtfB-DELTAN-DELTAV from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Limosilactobacillus reuteri
2.4.1.362 recombinant C-terminally His-tagged enzyme mutant GtfZ-CD2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Apilactobacillus kunkeei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.B34 amylose V + H2O hydrolytic activity, amylose V with Mw 170 kDa Limosilactobacillus reuteri ?
-
 ?
2.4.1.B34 amylose V + H2O hydrolytic activity, amylose V with Mw 170 kDa Limosilactobacillus reuteri 121 ?
-
 ?
2.4.1.B34 amylose V + maltose transglycosylation activity, amylose V with 170 kDa Limosilactobacillus reuteri ?
-
 ?
2.4.1.B34 amylose V + maltose transglycosylation activity, amylose V with 170 kDa Limosilactobacillus reuteri 121 ?
-
 ?
2.4.1.B34 additional information the enzyme performs transglycosylation and/or hydrolytic activity. The linear alpha(1->4)-linked glucose units disappear and linear alpha(1->6)-linked glucose units appear. The total enzyme activity of GtfB-DELTAN-DELTAV is determined by the amylose-iodine staining method. Synthesis of isomalto/malto-polysaccharides (IMMP) from starch. Reaction product analyses, the IMMP product consists of terminal, and 6-substituted glucosyl units, overview. IMMP with 18.3 kDa Limosilactobacillus reuteri ?
-
-
2.4.1.B34 additional information the enzyme performs transglycosylation and/or hydrolytic activity. The linear alpha(1->4)-linked glucose units disappear and linear alpha(1->6)-linked glucose units appear. The total enzyme activity of GtfB-DELTAN-DELTAV is determined by the amylose-iodine staining method. Synthesis of isomalto/malto-polysaccharides (IMMP) from starch. Reaction product analyses, the IMMP product consists of terminal, and 6-substituted glucosyl units, overview. IMMP with 18.3 kDa Limosilactobacillus reuteri 121 ?
-
-
2.4.1.362 dextran + sucrose dextran Mw is 70 kDa, transglucosylation activity Apilactobacillus kunkeei D-fructose + dextran containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 dextran + sucrose dextran Mw is 70 kDa, transglucosylation activity Apilactobacillus kunkeei DSM 12361 D-fructose + dextran containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 isomalto/malto-polysaccharide + sucrose alpha(1->3)-branched IMMP, IMMP Mw is 70 kDa, transglucosylation activity Apilactobacillus kunkeei D-fructose + isomalto/malto-polysaccharide containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 additional information enzyme GtfZ-D2 produces partially branched polymers. Analysis of the glucosyl linkage composition of the (alpha1->3)-branched polymers. Reaction product analyses with 1D 1H NMR, overview. GtfZ-CD2 of Lactobacillus kunkeei uses sucrose as a glucose donor to decorate dextran molecules, adding single (alpha1->3)-branched glucose units on the linear (alpha1->6) glucose chain. GtfZ-CD2 also exhibits (alpha1->3)-branching activity on IMMP (a product of reaction of GtfB, EC 2.4.1.B52, with about 90% (alpha1->6)-linkages) Apilactobacillus kunkeei ?
-
-
2.4.1.362 additional information enzyme GtfZ-D2 produces partially branched polymers. Analysis of the glucosyl linkage composition of the (alpha1->3)-branched polymers. Reaction product analyses with 1D 1H NMR, overview. GtfZ-CD2 of Lactobacillus kunkeei uses sucrose as a glucose donor to decorate dextran molecules, adding single (alpha1->3)-branched glucose units on the linear (alpha1->6) glucose chain. GtfZ-CD2 also exhibits (alpha1->3)-branching activity on IMMP (a product of reaction of GtfB, EC 2.4.1.B52, with about 90% (alpha1->6)-linkages) Apilactobacillus kunkeei DSM 12361 ?
-
-
2.4.1.362 sucrose + (1->6)-alpha-D-glucan
-
 Apilactobacillus kunkeei D-fructose + (1->6)-alpha-D-glucan containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 sucrose + (1->6)-alpha-D-glucan transglucosylation activity Apilactobacillus kunkeei D-fructose + (1->6)-alpha-D-glucan containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 sucrose + (1->6)-alpha-D-glucan
-
 Apilactobacillus kunkeei DSM 12361 D-fructose + (1->6)-alpha-D-glucan containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 sucrose + (1->6)-alpha-D-glucan transglucosylation activity Apilactobacillus kunkeei DSM 12361 D-fructose + (1->6)-alpha-D-glucan containing a (1->3)-alpha-D-glucose branch
-
 ?
2.4.1.362 sucrose + H2O sucrose hydrolysis Apilactobacillus kunkeei D-fructose + D-glucose
-
 ?
2.4.1.362 sucrose + H2O sucrose hydrolysis Apilactobacillus kunkeei DSM 12361 D-fructose + D-glucose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.B34 GtfB
-
 Limosilactobacillus reuteri
2.4.1.B34 UDP-N-acetylglucosamine-peptide N-acetylglucosaminyltransferase stabilizing protein UniProt Limosilactobacillus reuteri
2.4.1.362 branching sucrase
-
 Apilactobacillus kunkeei
2.4.1.362 GtfZ
-
 Apilactobacillus kunkeei

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.B34 37 40 assay at Limosilactobacillus reuteri
2.4.1.362 30
-
 assay at Apilactobacillus kunkeei

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.B34 95
-
 purified recombinant enzyme, 20 min, inactivation Limosilactobacillus reuteri
2.4.1.362 95
-
 purified recombinant enzyme, 20 min, inactivation Apilactobacillus kunkeei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.B34 5
-
 assay at Limosilactobacillus reuteri
2.4.1.362 5.5
-
 assay at Apilactobacillus kunkeei

General Information

EC Number General Information Comment Organism
2.4.1.B34 physiological function identification of 4,6-alpha-glucanotransferase enzymes of the glycosyl hydrolase (GH) family 70 (GH70) that cleave alpha(1->4)-linkages in amylose and introduce alpha(1->6)-linkages in linear chains. The 4,6-alpha-glucanotransferase of Lactobacillus reuteri strain 121 converts amylose into an isomalto/malto-polysaccharide (IMMP) with 90% alpha(1->6)-linkages Limosilactobacillus reuteri