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Literature summary extracted from

  • Kim, Y.; Yeom, S.; Oh, D.
    Reduction of galactose inhibition via the mutation of beta-galactosidase from Caldicellulosiruptor saccharolyticus for lactose hydrolysis (2011), Biotechnol. Lett., 33, 353-358 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.23 expressed in Escherichia coli ER2566 Caldicellulosiruptor saccharolyticus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.23 E359A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The E359A mutant exhibits no activity Caldicellulosiruptor saccharolyticus
3.2.1.23 F349A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The F349A, R111A, and W187A mutant enzyme in the presence of galactose shows remaining activity of 65% Caldicellulosiruptor saccharolyticus
3.2.1.23 F349S mutant exhibits the highest activity in the presence of galactose relative to the activity measured in the absence of galactose among the tested mutant enzymes at position 349. The Ki-value for galactose of the F349S mutant (160 mM), which is 13fold that of the wild-type enzyme, is the highest among the reported values of beta-galactosidase. The wild-type enzyme hydrolyzes 62% of 100 g lactose/l with the addition of 30 g galactose/l, whereas the F349S mutant hydrolyzes more than 99% Caldicellulosiruptor saccharolyticus
3.2.1.23 H362A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The H362A mutant in the presence of galactose exhibits remaining activity below 30% Caldicellulosiruptor saccharolyticus
3.2.1.23 I44A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The I44A mutant in the presence of galactose exhibits remaining activity below 30% Caldicellulosiruptor saccharolyticus
3.2.1.23 N149A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The N149A mutant in the presence of galactose exhibits remaining activity below 30% Caldicellulosiruptor saccharolyticus
3.2.1.23 R111A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The F349A, R111A, and W187A mutant enzyme in the presence of galactose shows remaining activity of 46% Caldicellulosiruptor saccharolyticus
3.2.1.23 S41A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The S41A mutant in the presence of galactose exhibits remaining activity below 30% Caldicellulosiruptor saccharolyticus
3.2.1.23 W187A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The F349A, R111A, and W187A mutant enzyme in the presence of galactose shows remaining activity of 44% Caldicellulosiruptor saccharolyticus
3.2.1.23 W319A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The W319A mutant in the presence of galactose exhibits remaining activity below 30% Caldicellulosiruptor saccharolyticus
3.2.1.23 Y281A 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The Y281A mutant exhibits no activity Caldicellulosiruptor saccharolyticus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.23 D-galactose 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The Y281A and E359A mutants exhibit no activity and the S41A, I44A, N149A, W319A, and H362A mutants in the presence of galactose exhibit remaining activity below 30%. The F349A, R111A, and W187A mutant enzymes in the presence of galactose show remaining activity of 65, 46, and 44%, respectively Caldicellulosiruptor saccharolyticus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.23 80000
-
-
 Caldicellulosiruptor saccharolyticus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.23 Caldicellulosiruptor saccharolyticus A4XI97
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.23
-
 Caldicellulosiruptor saccharolyticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.23 4-nitrophenyl beta-D-galactopyranoside + H2O
-
 Caldicellulosiruptor saccharolyticus 4-nitrophenol + beta-D-galactopyranose
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.23 80
-
 assay at Caldicellulosiruptor saccharolyticus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.23 12
-
 D-galactose pH 6.0, 80°C, wild-type enzyme Caldicellulosiruptor saccharolyticus
3.2.1.23 70
-
 D-galactose pH 6.0, 80°C, mutant enzyme F349V Caldicellulosiruptor saccharolyticus
3.2.1.23 147
-
 D-galactose pH 6.0, 80°C, mutant enzyme F349L Caldicellulosiruptor saccharolyticus
3.2.1.23 156
-
 D-galactose pH 6.0, 80°C, mutant enzyme F349A Caldicellulosiruptor saccharolyticus
3.2.1.23 160
-
 D-galactose pH 6.0, 80°C, mutant enzyme F349S Caldicellulosiruptor saccharolyticus