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Literature summary for 3.2.1.55 extracted from

  • Koseki, T.; Miwa, Y.; Mese, Y.; Miyanaga, A.; Fushinobu, S.; Wakagi, T.; Shoun, H.; Matsuzawa, H.; Hashizume, K.
    Mutational analysis of N-glycosylation recognition sites on the biochemical properties of Aspergillus kawachii alpha-L-arabinofuranosidase 54 (2006), Biochim. Biophys. Acta, 1760, 1458-1464.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Pichia pastori as an active enzyme accumulated in the culture broth, wild-type and mutant enzymes lacking one or both asparagine residues in the N-glycosylation sites N83Q, N202Q and N83Q/N202Q Aspergillus luchuensis

Protein Variants

Protein Variants Comment Organism
N202Q T50-value is 3°C lower than wild-type value. Vmax towards p-nitrophenyl-alpha-L-arabinofuranoside is 0.76fold of wild-type value. No significant effect on kcat/Km with p-nitrophenyl-alpha-L-arabinofuranoside as substrate. Significant decrease in specific activity towards arabinan and debranched arabinan compared to wild-type enzyme Aspergillus luchuensis
N83Q T50-value is 1°C lower than wild-type value. No significant effect on kcat/Km Aspergillus luchuensis
N83Q/N202Q T50-value is 3°C lower than wild-type valueVmax towards p-nitrophenyl-alpha-L-arabinofuranoside is 0.71fold of wild-type value. No significant effect on kcat/K with p-nitrophenyl-alpha-L-arabinofuranoside as substrate. Significant decrease in specific activity towards arabinan and debranched arabinan compared to wild-type enzyme Aspergillus luchuensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.69
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, mutant enzyme N83Q/N202Q Aspergillus luchuensis
0.86
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, mutant enzyme N202Q Aspergillus luchuensis
1.04
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, wild-type enzyme Aspergillus luchuensis
1.07
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, mutant enzyme N83Q Aspergillus luchuensis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
59000
-
x * 59000, mutant N83Q/N202Q, SDS-PAGE Aspergillus luchuensis
60700
-
x * 60700, mutant N83Q, SDS-PAGE Aspergillus luchuensis
62000
-
x * 62000, mutant N202Q, SDS-PAGE Aspergillus luchuensis
64500
-
x * 64500, wild-type enzyme, SDS-PAGE Aspergillus luchuensis

Organism

Organism UniProt Comment Textmining
Aspergillus luchuensis
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein the native enzyme contains N-linked oligosaccharides. The Asn202-linked oligosaccharide may contribute to thermostability Aspergillus luchuensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
arabinan + H2O
-
Aspergillus luchuensis ?
-
?
debranched arabinan + H2O
-
Aspergillus luchuensis ?
-
?
p-nitrophenyl-alpha-L-arabinofuranoside + H2O
-
Aspergillus luchuensis p-nitrophenol + L-arabinofuranose
-
?

Subunits

Subunits Comment Organism
? x * 59000, mutant N83Q/N202Q, SDS-PAGE Aspergillus luchuensis
? x * 60700, mutant N83Q, SDS-PAGE Aspergillus luchuensis
? x * 62000, mutant N202Q, SDS-PAGE Aspergillus luchuensis
? x * 64500, wild-type enzyme, SDS-PAGE Aspergillus luchuensis

Synonyms

Synonyms Comment Organism
AkAbf54
-
Aspergillus luchuensis
alpha-L-arabinofuranosidase 54
-
Aspergillus luchuensis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
T50-value of mutant enzyme N202Q is 60.1°C Aspergillus luchuensis
60
-
T50-value of mutant enzyme N83Q/N202Q is 60.1°C Aspergillus luchuensis
62
-
T50-value of mutant enzyme N83Q is 62.2°C Aspergillus luchuensis
63
-
T50-value of wild-type enzyme is 63.3°C Aspergillus luchuensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0694
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, mutant enzyme N83Q/N202Q Aspergillus luchuensis
0.0755
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, mutant enzyme N202Q Aspergillus luchuensis
0.0898
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, mutant enzyme N83Q Aspergillus luchuensis
0.099
-
p-nitrophenyl-alpha-L-arabinofuranoside pH 4.0, 37°C, wild-type enzyme Aspergillus luchuensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4
-
hydrolytic activity of wild-type enzyme Aspergillus luchuensis

pH Stability

pH Stability pH Stability Maximum Comment Organism
3 7 stable Aspergillus luchuensis