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Literature summary for 3.2.1.23 extracted from

  • Hildebrandt, P.; Wanarska, M.; Kur, J.
    A new cold-adapted beta-D-galactosidase from the Antarctic Arthrobacter sp. 32c - gene cloning, overexpression, purification and properties (2009), BMC Microbiol., 9, 151.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
ethanol the relative enzyme activity is increasing up to 120% in the presence of 8% (v/v) ethanol at pH 5.5 Arthrobacter sp.

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli TOP10F' cells and Pichia pastoris strain GS115 Arthrobacter sp.

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ the enzyme activity is completely inhibited by 5 mM Cu2+ Arthrobacter sp.
Fe2+ 25% residual activity at 5 mM Fe2+ Arthrobacter sp.
Mn2+ 11% residual activity at 5 mM Mn2+ Arthrobacter sp.
additional information the activity of the beta-D-galactosidase is not considerably affected by ditiothreitol, beta-mercaptoethanol, and L-cysteine Arthrobacter sp.
Ni2+ 38% residual activity at 5 mM Ni2+ Arthrobacter sp.
reduced glutathione 6% residual activity at 5 mM Arthrobacter sp.
Zn2+ the enzyme activity is completely inhibited by 5 mM Zn2+ Arthrobacter sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.62
-
2-nitrophenyl-beta-D-galactopyranoside at 50°C Arthrobacter sp.
3.15
-
2-nitrophenyl-beta-D-galactopyranoside at 40°C Arthrobacter sp.
3.46
-
2-nitrophenyl-beta-D-galactopyranoside at 30°C Arthrobacter sp.
4.86
-
2-nitrophenyl-beta-D-galactopyranoside at 20°C Arthrobacter sp.
5.11
-
2-nitrophenyl-beta-D-galactopyranoside at 55°C Arthrobacter sp.
5.75
-
2-nitrophenyl-beta-D-galactopyranoside at 10°C Arthrobacter sp.
39.73
-
lactose at 50°C Arthrobacter sp.
44.31
-
lactose at 40°C Arthrobacter sp.
53.67
-
lactose at 30°C Arthrobacter sp.
67.82
-
lactose at 20°C Arthrobacter sp.
77.54
-
lactose at 10°C Arthrobacter sp.

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the activity is not considerably affected by Na+, K+, Mg2+, Co2+, and Ca2+ (5 mM) Arthrobacter sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
70000
-
3 * 70000, SDS-PAGE Arthrobacter sp.
76140
-
subunit, calculated from amino acid sequence Arthrobacter sp.
195600
-
gel filtration Arthrobacter sp.

Organism

Organism UniProt Comment Textmining
Arthrobacter sp. C7ASJ5
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation and p-aminobenzyl-1-thio-beta-D-galactopyranoside-agarose column chromatography Arthrobacter sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-nitrophenyl beta-D-galactopyranoside + H2O the enzyme displays 4times higher level of activity with 4-nitrophenyl beta-D-galactopyranoside than with 2-nitrophenyl beta-D-galactopyranoside Arthrobacter sp. 2-nitrophenol + beta-D-galactopyranose
-
?
2-nitrophenyl beta-D-glucopyranoside + H2O 0.5% of the activity with 2-nitrophenyl beta-D-galactopyranoside Arthrobacter sp. 2-nitrophenol + D-glucopyranose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O the enzyme displays 4times higher level of activity with 4-nitrophenyl beta-D-galactopyranoside than with 2-nitrophenyl beta-D-galactopyranoside Arthrobacter sp. 4-nitrophenol + beta-D-galactopyranose
-
?
4-nitrophenyl beta-D-glucopyranoside + H2O 1.4% of the activity with 2-nitrophenyl beta-D-galactopyranoside Arthrobacter sp. 4-nitrophenol + D-glucopyranose
-
?
lactose + H2O
-
Arthrobacter sp. D-galactose + D-glucose
-
?

Subunits

Subunits Comment Organism
homotrimer 3 * 70000, SDS-PAGE Arthrobacter sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
Arthrobacter sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
-
25 60% of the maximum activity of the enzyme is determined at 25°C and 15% of the maximum activity is detected at 0°C, the enzyme is active in the range of temperature 4-8°C Arthrobacter sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.76
-
lactose at 10°C Arthrobacter sp.
2.36
-
lactose at 20°C Arthrobacter sp.
4.81
-
lactose at 30°C Arthrobacter sp.
5.73
-
lactose at 40°C Arthrobacter sp.
6.98
-
lactose at 50°C Arthrobacter sp.
52.4
-
2-nitrophenyl-beta-D-galactopyranoside at 10°C Arthrobacter sp.
71.2
-
2-nitrophenyl-beta-D-galactopyranoside at 55°C Arthrobacter sp.
81
-
2-nitrophenyl-beta-D-galactopyranoside at 20°C Arthrobacter sp.
123.9
-
2-nitrophenyl-beta-D-galactopyranoside at 30°C Arthrobacter sp.
169.9
-
2-nitrophenyl-beta-D-galactopyranoside at 40°C Arthrobacter sp.
212.4
-
2-nitrophenyl-beta-D-galactopyranoside at 50°C Arthrobacter sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Arthrobacter sp.

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8.5 over 90% of the maximum activity in the pH range of 6.5-8.5 Arthrobacter sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 7.5 over 50% of activity at pH 5.5-7.5 at 30°C Arthrobacter sp.

pI Value

Organism Comment pI Value Maximum pI Value
Arthrobacter sp. calculated from amino acid sequence
-
5.6

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.023
-
lactose at 10°C Arthrobacter sp.
0.035
-
lactose at 20°C Arthrobacter sp.
0.091
-
lactose at 30°C Arthrobacter sp.
0.129
-
lactose at 40°C Arthrobacter sp.
0.176
-
lactose at 50°C Arthrobacter sp.
9.12
-
2-nitrophenyl-beta-D-galactopyranoside at 10°C Arthrobacter sp.
13.93
-
2-nitrophenyl-beta-D-galactopyranoside at 55°C Arthrobacter sp.
16.67
-
2-nitrophenyl-beta-D-galactopyranoside at 20°C Arthrobacter sp.
35.81
-
2-nitrophenyl-beta-D-galactopyranoside at 30°C Arthrobacter sp.
53.92
-
2-nitrophenyl-beta-D-galactopyranoside at 40°C Arthrobacter sp.
81.07
-
2-nitrophenyl-beta-D-galactopyranoside at 50°C Arthrobacter sp.