Application | Comment | Organism |
---|---|---|
industry | the beta-galactosidase from Kluyveromyces lactis is a protein of outstanding biotechnological interest in the food industry and milk whey reutilization, optimization of extraction and downstream processing of the intracellular enzyme for reduction of costs in industrial production by genetic modification, overview | Kluyveromyces lactis |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and recombinant hybrid mutant enzymes in Escherichia coli strain DH5alpha | Aspergillus niger |
expression of wild-type and recombinant hybrid mutant enzymes in Escherichia coli strain DH5alpha | Kluyveromyces lactis |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a hybrid mutant enzyme exchanging domains from Kluyveromyces lactis and Aspergillus niger enzymes, addition of a secretory signal in the N-terminus of the recombinant protein, enzyme optimization for industrial productions by fusion with extracellular and more stable enzyme from Aspergillus niger, overview, kinetics of growth and secretion | Kluyveromyces lactis |
additional information | construction of a hybrid mutant enzyme exchanging domains from Kluyveromyces lactis and Aspergillus niger enzymes, addition of a secretory signal in the N-terminus of the recombinant protein, Kluyveromyces lactis enzyme optimization for industrial productions by fusion with extracellular and more stable enzyme from Aspergillus niger, overview, kinetics of growth and secretion | Aspergillus niger |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | slight inhibition of both the wild-type Kluyveromyces lactis enzyme and the recombinant hybrid enzyme | Kluyveromyces lactis | |
Mn2+ | activates the recombinant hybrid enzyme, but inhibits the wild-type Kluyveromyces lactis enzyme | Kluyveromyces lactis | |
Ni2+ | activates the recombinant hybrid enzyme, but inhibits the wild-type Kluyveromyces lactis enzyme | Kluyveromyces lactis | |
Zn2+ | slightly inhibits the recombinant hybrid enzyme | Aspergillus niger | |
Zn2+ | slightly activates the wild-type Kluyveromyces lactis enzyme, but slightly inhibits the recombinant hybrid enzyme | Kluyveromyces lactis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Aspergillus niger | |
additional information | - |
additional information | kinetics | Kluyveromyces lactis | |
0.8 | - |
2-nitrophenyl-beta-D-galactopyranoside | pH 6.5, 40°C, recombinant hybrid enzyme | Aspergillus niger | |
0.8 | - |
2-nitrophenyl-beta-D-galactopyranoside | pH 6.5, 40°C, recombinant hybrid enzyme | Kluyveromyces lactis | |
1.5 | - |
2-nitrophenyl-beta-D-galactopyranoside | pH 6.5, 40°C, wild-type enzyme | Kluyveromyces lactis | |
8.7 | - |
lactose | pH 6.5, 40°C, recombinant hybrid enzyme | Aspergillus niger | |
8.7 | - |
lactose | pH 6.5, 40°C, recombinant hybrid enzyme | Kluyveromyces lactis | |
21 | - |
lactose | pH 6.5, 40°C, wild-type enzyme | Kluyveromyces lactis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Aspergillus niger | - |
- |
intracellular | - |
Kluyveromyces lactis | 5622 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | activation of both the wild-type Kluyveromyces lactis enzyme and the recombinant hybrid enzyme | Kluyveromyces lactis | |
Mg2+ | activation of the recombinant hybrid enzyme | Aspergillus niger | |
Mn2+ | activates the recombinant hybrid enzyme | Aspergillus niger | |
Mn2+ | activates the recombinant hybrid enzyme, but inhibits the wild-type Kluyveromyces lactis enzyme | Kluyveromyces lactis | |
Ni2+ | activates the recombinant hybrid enzyme | Aspergillus niger | |
Ni2+ | activates the recombinant hybrid enzyme, but inhibits the wild-type Kluyveromyces lactis enzyme | Kluyveromyces lactis | |
Zn2+ | slightly activates the wild-type Kluyveromyces lactis enzyme, but slightly inhibits the recombinant hybrid enzyme | Kluyveromyces lactis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
lactose + H2O | Aspergillus niger | - |
D-glucose + D-galactose | - |
? | |
lactose + H2O | Kluyveromyces lactis | - |
D-glucose + D-galactose | - |
? | |
lactose + H2O | Kluyveromyces lactis MW 190-9B | - |
D-glucose + D-galactose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | - |
- |
- |
Kluyveromyces lactis | - |
- |
- |
Kluyveromyces lactis MW 190-9B | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
beta-D-galactopyranosyl-(1-4)-beta-D-galactopyranosyl-(1-6)-beta-D-galactopyranosyl-(1-3)-beta-D-galactopyranose + 3 H2O = 4 beta-D-galactopyranose | the conserved residues E482, M522, Y523 and E551 are important in catalysis | Kluyveromyces lactis |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | kinetics of growth and secretion | Aspergillus niger | - |
additional information | liquid batch cultures, kinetics of growth and secretion | Kluyveromyces lactis | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-nitrophenyl beta-D-galactopyranoside + H2O | - |
Aspergillus niger | 2-nitrophenol + beta-D-galactose | - |
? | |
2-nitrophenyl beta-D-galactopyranoside + H2O | - |
Kluyveromyces lactis | 2-nitrophenol + beta-D-galactose | - |
? | |
2-nitrophenyl beta-D-galactopyranoside + H2O | - |
Kluyveromyces lactis MW 190-9B | 2-nitrophenol + beta-D-galactose | - |
? | |
lactose + H2O | - |
Aspergillus niger | D-glucose + D-galactose | - |
? | |
lactose + H2O | - |
Kluyveromyces lactis | D-glucose + D-galactose | - |
? | |
lactose + H2O | - |
Kluyveromyces lactis MW 190-9B | D-glucose + D-galactose | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | tertiary structure of the beta-galactosidase of Kluyveromyces lactis and the recombinant hybrid enzyme, protein structure homology-modelling, overview | Kluyveromyces lactis |
More | tertiary structure of the recombinant hybrid enzyme, protein structure homology-modelling, overview | Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
wild-type enzyme | Kluyveromyces lactis |
40 | - |
recombinant mutant hybrid enzyme | Aspergillus niger |
40 | - |
recombinant mutant hybrid enzyme | Kluyveromyces lactis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
30 min, recombinant hybrid enzyme, stable | Aspergillus niger |
30 | - |
30 min, recombinant hybrid enzyme, stable, the wild-type enzyme loses 45% activity within 60 min | Kluyveromyces lactis |
40 | - |
15 min, recombinant hybrid enzyme, stable | Aspergillus niger |
40 | - |
15 min, recombinant hybrid enzyme, stable, the wild-type enzyme loses 10% activity within 10 min | Kluyveromyces lactis |
50 | - |
15 min, recombinant hybrid enzyme, loss of 40% activity | Aspergillus niger |
50 | - |
15 min, recombinant hybrid enzyme, loss of 40% activity, the wild-type enzyme loses 60% activity | Kluyveromyces lactis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
recombinant mutant hybrid enzyme | Aspergillus niger |
6.5 | - |
recombinant mutant hybrid enzyme | Kluyveromyces lactis |
7 | - |
wild-type enzyme | Kluyveromyces lactis |