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Literature summary for 3.2.1.20 extracted from

  • Tsujimoto, Y.; Tanaka, H.; Takemura, R.; Yokogawa, T.; Shimonaka, A.; Matsui, H.; Kashiwabara, S.; Watanabe, K.; Suzuki, Y.
    Molecular determinants of substrate recognition in thermostable alpha-glucosidases belonging to glycoside hydrolase family 13 (2007), J. Biochem., 142, 87-93.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Geobacillus stearothermophilus

Protein Variants

Protein Variants Comment Organism
A200V site-directed mutagenesis, the mutant is almost inactive Geobacillus stearothermophilus
A200V/S202N/H203M site-directed mutagenesis, the mutant shows altered substrate specificity and highly reduced activity compared to the wild-type enzyme Geobacillus stearothermophilus
N258P site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme with reduced activity against maltose and increased activity against isomaltose Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
maltooligosaccharides + H2O Geobacillus stearothermophilus
-
alpha-D-glucose
-
?
maltooligosaccharides + H2O Geobacillus stearothermophilus ATCC 12016
-
alpha-D-glucose
-
?
maltose + H2O Geobacillus stearothermophilus
-
alpha-D-glucose + D-glucose
-
?
maltose + H2O Geobacillus stearothermophilus ATCC 12016
-
alpha-D-glucose + D-glucose
-
?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
strain ATCC12016
-
Geobacillus stearothermophilus ATCC 12016
-
strain ATCC12016
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109 by heat treatment at 55°C for 10 min, followed by anion exchange, adsorption, hydrophobic interaction chromatography, and gel filtration Geobacillus stearothermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.7
-
purified recombinant mutant A200V, substrate maltose Geobacillus stearothermophilus
3280
-
purified recombinant mutant N258P, substrate maltose Geobacillus stearothermophilus
6890
-
purified recombinant wild-type enzyme, substrate maltose Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl alpha-D-glucopyranoside + H2O
-
Geobacillus stearothermophilus 4-nitrophenol + alpha-D-glucose
-
?
4-nitrophenyl alpha-D-glucopyranoside + H2O
-
Geobacillus stearothermophilus ATCC 12016 4-nitrophenol + alpha-D-glucose
-
?
isomaltose + H2O
-
Geobacillus stearothermophilus 2 alpha-D-glucose
-
?
isomaltose + H2O
-
Geobacillus stearothermophilus ATCC 12016 2 alpha-D-glucose
-
?
maltooligosaccharides + H2O
-
Geobacillus stearothermophilus alpha-D-glucose
-
?
maltooligosaccharides + H2O
-
Geobacillus stearothermophilus ATCC 12016 alpha-D-glucose
-
?
maltose + H2O
-
Geobacillus stearothermophilus alpha-D-glucose + D-glucose
-
?
maltose + H2O
-
Geobacillus stearothermophilus ATCC 12016 alpha-D-glucose + D-glucose
-
?
additional information the enzyme specifically hydrolyzes alpha-1,4-glucosidic bonds in maltose, maltooligosaccharides, and alpha-glucans, isomaltose is a poor substrate, residues Ala200 and Asn258 in the conserved region II and III, respectively, are largely responsible for substrate recognition Geobacillus stearothermophilus ?
-
?
additional information the enzyme specifically hydrolyzes alpha-1,4-glucosidic bonds in maltose, maltooligosaccharides, and alpha-glucans, isomaltose is a poor substrate, residues Ala200 and Asn258 in the conserved region II and III, respectively, are largely responsible for substrate recognition Geobacillus stearothermophilus ATCC 12016 ?
-
?

Subunits

Subunits Comment Organism
More the enzyme contains four conserved regions I-IV Geobacillus stearothermophilus

Synonyms

Synonyms Comment Organism
alpha-1,4-glucosidase
-
Geobacillus stearothermophilus
More the enzyme belongs to the glycoside hydrolase family 13 Geobacillus stearothermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55 60 assay at Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.8 7 assay at Geobacillus stearothermophilus