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Literature summary for 2.4.1.230 extracted from

  • Nakai, H.; Petersen, B.O.; Westphal, Y.; Dilokpimol, A.; Abou Hachem, M.; Duus, J.?.; Schols, H.A.; Svensson, B.
    Rational engineering of Lactobacillus acidophilus NCFM maltose phosphorylase into either trehalose or kojibiose dual specificity phosphorylase (2010), Protein Eng. Des. Sel., 23, 781-787.
    View publication on PubMed
Search for more literature for 2.4.1.230

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged LaMP loop 3 mutants in Escherichia coli strain BL21(DE3) Lactobacillus acidophilus

Protein Variants

Protein Variants Comment Organism
additional information substitution of maltose phosphorylase, EC 2.4.1.8, His413-Glu421, His413-Ile418 and His413-Glu415 from loop 3, that include His413 and Glu415 presumably recognising the alpha-anomeric O-1 group of the glucose moiety at subsite 1+, by corresponding to segments from Ser426-Ala431 in trehalose phosphorylase, EC 2.4.1.64, and Thr419-Phe427 in kojibiose phosphorylase, EC 2.4.1.230, thus confers the maltose phosphorylase with phosphorolytic activity towards trehalose and kojibiose, respectively Lactobacillus acidophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
9.3
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 3 of maltose phosphorylase Lactobacillus acidophilus
16
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 2 of maltose phosphorylase Lactobacillus acidophilus
84
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 1 of maltose phosphorylase Lactobacillus acidophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-alpha-D-glucosyl-D-glucose + phosphate Thermoanaerobacter brockii
-
 D-glucose + beta-D-glucose 1-phosphate
-
 ?
2-alpha-D-glucosyl-D-glucose + phosphate Thermoanaerobacter brockii ATCC 35047
-
 D-glucose + beta-D-glucose 1-phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Lactobacillus acidophilus
-
-
-
Thermoanaerobacter brockii
-
-
-
Thermoanaerobacter brockii ATCC 35047
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged LaMP loop 3 mutants from Escherichia coli strain BL21(DE3) by affinity chromatography and dialysis Lactobacillus acidophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-alpha-D-glucosyl-D-glucose + phosphate
-
 Thermoanaerobacter brockii D-glucose + beta-D-glucose 1-phosphate
-
 ?
2-alpha-D-glucosyl-D-glucose + phosphate activity of a LaMP loop 3 maltose phosphorylase mutant, no activity with wild-type maltose phosphorylase, which is strictly specific for maltose Lactobacillus acidophilus D-glucose + beta-D-glucose 1-phosphate
-
 r
2-alpha-D-glucosyl-D-glucose + phosphate activity of a LaMP loop 3 maltose phosphorylase mutant, no activity with wild-type maltose phosphorylase, which is strictly specific for maltose Lactobacillus acidophilus NCFM D-glucose + beta-D-glucose 1-phosphate
-
 r
2-alpha-D-glucosyl-D-glucose + phosphate
-
 Thermoanaerobacter brockii ATCC 35047 D-glucose + beta-D-glucose 1-phosphate
-
 ?

Synonyms

Synonyms Comment Organism
kojibiose phosphorylase
-
 Thermoanaerobacter brockii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 phosphorolysis and reverse phosphorolysis Lactobacillus acidophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.097
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 1 of maltose phosphorylase Lactobacillus acidophilus
0.12
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 2 of maltose phosphorylase Lactobacillus acidophilus
2.4
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 3 of maltose phosphorylase Lactobacillus acidophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4
-
 reverse phosphorolysis Lactobacillus acidophilus
6.2
-
 phosphorolysis Lactobacillus acidophilus

General Information

General Information Comment Organism
evolution the enzyme is a member of glycoside hydrolase family 65, GH65 Lactobacillus acidophilus
evolution the enzyme is a member of glycoside hydrolase family 65, GH65 Thermoanaerobacter brockii
additional information structure-function relationship analysis, homology modelling, overview. Substitution of maltose phosphorylase, EC 2.4.1.8, His413-Glu421, His413-Ile418 and His413-Glu415 from loop 3, that include His413 and Glu415 presumably recognising the alpha-anomeric O-1 group of the glucose moiety at subsite 1+, by corresponding to segments from Ser426-Ala431 in trehalose phosphorylase, EC 2.4.1.64, and Thr419-Phe427 in kojibiose phosphorylase, EC 2.4.1.230, thus confers the maltose phosphorylase with phosphorolytic activity towards trehalose and kojibiose, respectively. The loop 3 in GH65 disaccharide phosphorylase is a key determinant for specificity both in phosphorolysis and in regiospecific reverse phosphorolysis Lactobacillus acidophilus
additional information structure-function relationship analysis, overview. The sequence Thr419-Phe427 is responsible for substrate specificity of the enzyme for kojibiose. The loop 3 in GH65 disaccharide phosphorylase is a key determinant for specificity both in phosphorolysis and in regiospecific reverse phosphorolysis Thermoanaerobacter brockii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0011
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 1 of maltose phosphorylase Lactobacillus acidophilus
0.0075
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 2 of maltose phosphorylase Lactobacillus acidophilus
0.26
-
 2-alpha-D-glucosyl-D-glucose pH 6.2, 45°C, recombinant loop 3 mutant 3 of maltose phosphorylase Lactobacillus acidophilus