Cloned (Comment) | Organism |
---|---|
expression of mutant enzymes in Aspergillus niger strain NW188, subcloning in Escherichia coli DH5alpha | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
D154A | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Aspergillus niger |
D154E | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Aspergillus niger |
D186N | site-directed mutagenesis, reduced activity and altered pH profile compared to the wild-type enzyme | Aspergillus niger |
D186N/D221N | site-directed mutagenesis, reduced activity and altered pH profile compared to the wild-type enzyme | Aspergillus niger |
D221N | site-directed mutagenesis, reduced activity and altered pH profile compared to the wild-type enzyme | Aspergillus niger |
K239N | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Aspergillus niger |
N109A | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Aspergillus niger |
R176A | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Aspergillus niger |
R176D | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Aspergillus niger |
R176K | site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme | Aspergillus niger |
R236A | site-directed mutagenesis, active site residue exchange, completely inactive mutant | Aspergillus niger |
R236K | site-directed mutagenesis, active site residue exchange, completely inactive mutant | Aspergillus niger |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes, influence of pH on the kinetics | Aspergillus niger |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
x * 45000, SDS-PAGE | Aspergillus niger |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | Q01172 | family 1 pectin lyase A | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | the wild-type enzyme is N-glycosylated | Aspergillus niger |
Purification (Comment) | Organism |
---|---|
native wild-type enzyme and recombinant mutant enzymes to homogeneity | Aspergillus niger |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
[6-O-methyl-alpha-D-GalpA]n = [6-O-methyl-alpha-D-GalpA]m + 4-deoxy-6-O-methyl-alpha-D-galact-4-enuronosy-[6-O-methyl-alpha-D-GalpA]n-m-1 | reaction mechanism, catalytic residues Arg236, Arg176, and Lys239 are essentially involved in catalysis | Aspergillus niger |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | pH driven conformational changes of the enzyme, wild-type and mutants | Aspergillus niger | ? | - |
? | |
partially methyl-esterified pectin | prepared from citrus and Aspergillus sp. | Aspergillus niger | ? | - |
? | |
pectin | citrus pectin | Aspergillus niger | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 45000, SDS-PAGE | Aspergillus niger |
Synonyms | Comment | Organism |
---|---|---|
family 1 pectin lyase A | - |
Aspergillus niger |
PL1A | - |
Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Aspergillus niger |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
mutant D186N | Aspergillus niger |
6 | - |
wild-type enzyme | Aspergillus niger |
8 | - |
mutants D221N and D186N/D221N | Aspergillus niger |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | - |
Aspergillus niger |