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Literature summary extracted from

  • Moeller, M.S.; Fredslund, F.; Majumder, A.; Nakai, H.; Poulsen, J.C.; Lo Leggio, L.; Svensson, B.; Abou Hachem, M.
    Enzymology and structure of the GH13_31 glucan 1,6-alpha-glucosidase that confers isomaltooligosaccharide utilization in the probiotic Lactobacillus acidophilus NCFM (2012), J. Bacteriol., 194, 4249-4259.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.70 gene LBA0264, DNA and amino acid sequence determination and analysis, genetic organization, sequence comparisons and phylogenetic tree, the enzyme is encoded in the maltooligosaccharide operon, semiquantitative PCR expression analysis, cloning in Escherichia coli strain TOP10, expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Lactobacillus acidophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.70 purified recombinant His-tagged enzyme, sitting drop vapour diffusion method, mixing of 16 mg/ml protein in 20 mM MES-NaOH, pH 6.5, 100 mM NaCl, and 2 mM CaCl2, with reservoir solution containing 20% glycerol, 16% PEG 8000, and 0.1 M MES, pH 6.5, in a 2:1 ratio, room temperature, X-ray diffraction structure determination and analysis at 2.05 A resolution, molecular replacement and modelling Lactobacillus acidophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.70 D-glucose product inhibition Lactobacillus acidophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.70 additional information
-
 additional information Michaelis-Menten kinetics Lactobacillus acidophilus
3.2.1.70 2.38
-
 4-nitrophenyl-alpha-D-glucopyranoside purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 3.9
-
 panose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 14.1
-
 isomaltotriose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 16.1
-
 isomaltotetraose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 22.5
-
 isomaltose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 29.1
-
 Dextran purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.70 Ca2+ stabilizes, a Ca2 is bound in domain A in a loop located just before the first helix in a binding site, the Ca2+ octahedral coordination shell comprises the side chains of Asp20, Asn22, Asp24, and Asp28, the carbonyl oxygen of Ile26, and a water molecule Lactobacillus acidophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.70 dextran + H2O Lactobacillus acidophilus
-
 D-glucose + ?
-
 ?
3.2.1.70 dextran + H2O Lactobacillus acidophilus NCFM
-
 D-glucose + ?
-
 ?
3.2.1.70 isomaltooligosaccharide + H2O Lactobacillus acidophilus degradation D-glucose + ?
-
 ?
3.2.1.70 isomaltose + H2O Lactobacillus acidophilus
-
 2 D-glucose
-
 ?
3.2.1.70 isomaltose + H2O Lactobacillus acidophilus NCFM
-
 2 D-glucose
-
 ?
3.2.1.70 isomaltotetraose + H2O Lactobacillus acidophilus
-
 ?
-
 ?
3.2.1.70 isomaltotetraose + H2O Lactobacillus acidophilus NCFM
-
 ?
-
 ?
3.2.1.70 isomaltotriose + H2O Lactobacillus acidophilus
-
 ?
-
 ?
3.2.1.70 isomaltotriose + H2O Lactobacillus acidophilus NCFM
-
 ?
-
 ?
3.2.1.70 panose + H2O Lactobacillus acidophilus
-
 D-glucose + ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.70 Lactobacillus acidophilus Q5FMB7 gene LBA0264
-
3.2.1.70 Lactobacillus acidophilus NCFM Q5FMB7 gene LBA0264
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.70 recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, anion exchange chromatography, and ultrafiltration, to homogeneity Lactobacillus acidophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.70 335
-
 purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.70 4-nitrophenyl-alpha-D-glucopyranoside + H2O
-
 Lactobacillus acidophilus 4-nitrophenol + D-glucose
-
 ?
3.2.1.70 dextran + H2O
-
 Lactobacillus acidophilus D-glucose + ?
-
 ?
3.2.1.70 dextran + H2O catalytic efficiency on dextran and dextran/panose Lactobacillus acidophilus D-glucose + ?
-
 ?
3.2.1.70 dextran + H2O
-
 Lactobacillus acidophilus NCFM D-glucose + ?
-
 ?
3.2.1.70 dextran + H2O catalytic efficiency on dextran and dextran/panose Lactobacillus acidophilus NCFM D-glucose + ?
-
 ?
3.2.1.70 isomaltooligosaccharide + H2O
-
 Lactobacillus acidophilus D-glucose + ?
-
 ?
3.2.1.70 isomaltooligosaccharide + H2O degradation Lactobacillus acidophilus D-glucose + ?
-
 ?
3.2.1.70 isomaltose + H2O
-
 Lactobacillus acidophilus 2 D-glucose
-
 ?
3.2.1.70 isomaltose + H2O
-
 Lactobacillus acidophilus NCFM 2 D-glucose
-
 ?
3.2.1.70 isomaltotetraose + H2O
-
 Lactobacillus acidophilus ?
-
 ?
3.2.1.70 isomaltotetraose + H2O
-
 Lactobacillus acidophilus NCFM ?
-
 ?
3.2.1.70 isomaltotriose + H2O
-
 Lactobacillus acidophilus ?
-
 ?
3.2.1.70 isomaltotriose + H2O
-
 Lactobacillus acidophilus NCFM ?
-
 ?
3.2.1.70 panose + H2O
-
 Lactobacillus acidophilus D-glucose + ?
-
 ?
3.2.1.70 panose + H2O catalytic efficiency on dextran/panose Lactobacillus acidophilus D-glucose + ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.70 GH13_31 glucan 1,6-alpha-glucosidase
-
 Lactobacillus acidophilus
3.2.1.70 LaGH13_31
-
 Lactobacillus acidophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.70 39
-
-
 Lactobacillus acidophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.70 37
-
 purified recombinant His6-tagged enzyme, pH 6.0, inactivation rate constant is 0.0001/min, T1/2 at 37°C is 4.8 days Lactobacillus acidophilus
3.2.1.70 50
-
 purified recombinant His6-tagged enzyme, pH 6.0, inactivation rate constant is 0.0152/min, T1/2 at 50°C is 46 min Lactobacillus acidophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.70 384
-
 Dextran purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 475
-
 isomaltotriose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 501
-
 isomaltotetraose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 517
-
 isomaltose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 597
-
 4-nitrophenyl-alpha-D-glucopyranoside purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 612
-
 panose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.70 5.5
-
-
 Lactobacillus acidophilus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.70 4 8 purified recombinant His6-tagged enzyme, over 50% activity remaining Lactobacillus acidophilus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.70 40
-
 D-glucose pH 6.0, 37°C, recombinant enzyme Lactobacillus acidophilus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.70 Lactobacillus acidophilus recombinant His-tagged enzyme, isoelectric focusing
-
 6.5

Expression

EC Number Organism Comment Expression
3.2.1.70 Lactobacillus acidophilus besides the isolated locus encoding a GH13_31 enzyme, the ABC transporter and another GH13 in the maltooligosaccharide operon are induced in response to IMO or maltotetraose suggesting coregulation of alpha-1,6- and alpha-1,4-glucooligosaccharide utilization loci in Lactobacillus acidophilus NCFM additional information
3.2.1.70 Lactobacillus acidophilus the enzyme is induced by isomaltooligosaccharides and maltotetraose up

General Information

EC Number General Information Comment Organism
3.2.1.70 evolution the enzyme belongs to the glycosyl hydrolase family 13 subfamily 31 Lactobacillus acidophilus
3.2.1.70 additional information phylogenetic and activity motif analysis, overview. Catalytic residues are Asp198 and Glu240, active site structure, overview Lactobacillus acidophilus
3.2.1.70 physiological function coregulation of alpha-1,6- and alpha-1,4-glucooligosaccharide utilization loci in the organism Lactobacillus acidophilus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.70 13.6
-
 Dextran purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 23
-
 isomaltose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 31
-
 isomaltotetraose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 34
-
 isomaltotriose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 157
-
 panose purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus
3.2.1.70 251
-
 4-nitrophenyl-alpha-D-glucopyranoside purified recombinant enzyme, pH 6.0, 37°C Lactobacillus acidophilus