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Literature summary for 3.2.1.21 extracted from

  • Kar, B.; Verma, P.; Patel, G.K.; Sharma, A.K.
    Molecular cloning, characterization and in silico analysis of a thermostable beta-glucosidase enzyme from Putranjiva roxburghii with a significant activity for cellobiose (2017), Phytochemistry, 140, 151-165 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
DMSO 10% (v/v), native enzyme is activated to 102.3% of control, recombinant enzyme is activated to 106.4% of control, beta-glucosidase activity Putranjiva roxburghii
EDTA 1 mM, native enzyme is activated to 110.7% of control, recombinant enzyme is activated to 108.2% of control, beta-glucosidase activity Putranjiva roxburghii
SDS 1 mg/ml, native enzyme is activated to 118.3% of control, recombinant enzyme is activated to 115.6% of control, beta-glucosidase activity Putranjiva roxburghii
Triton X-100 10 mg/ml, native enzyme is activated to 113.4% of control, recombinant enzyme is activated to 111.7% of control, beta-glucosidase activity Putranjiva roxburghii

Application

Application Comment Organism
industry potentially an important industrial enzyme due to the broad specificity, catalytic efficiency and thermostability Putranjiva roxburghii

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli Putranjiva roxburghii

Protein Variants

Protein Variants Comment Organism
E173N mutant enzyme shows no activity Putranjiva roxburghii
E389N mutant enzyme shows no activity Putranjiva roxburghii
E446N beta-fucosidase activity is 26.49% compared to wild-type activity, beta-glucosidase activity is 21.37% compared to wild-type activity, beta-galactosidase activity is 15.7% compared to wild-type activity Putranjiva roxburghii
N172A beta-fucosidase activity is 18.38% compared to wild-type activity, beta-glucosidase activity is 17.65% compared to wild-type activity, beta-galactosidase activity is 13.4% compared to wild-type activity Putranjiva roxburghii

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol 1 mM, 9% loss of activity of native enzyme, 10% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
Cr3+ 1 mM, 29% loss of activity of native enzyme, 30% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
delta-gluconolactone 5 mM, 85% loss of activity of native enzyme, 86% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
DTT 1 mM, 4% loss of activity of native enzyme, 5% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
Guanidine HCl 10 mM, 8% loss of activity of native enzyme, 10% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
Hg2+ 1 mM, 55% loss of activity of native enzyme, 58% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
Mg2+ 1 mM, 7% loss of activity, native and recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
additional information significant resistant to D-glucose concentration up to 500 mM Putranjiva roxburghii
N-bromosuccinamide 2 mM, 92% loss of activity of native enzyme, 94% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
Ni2+ 1 mM, 16% loss of activity of native enzyme, 18% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
Zn2+ 1 mM, 47% loss of activity of native enzyme, 52% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.46
-
4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
0.51
-
4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
0.53
-
4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
0.58
-
4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
0.64
-
4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
0.68
-
4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
0.81
-
cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
0.87
-
cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ 1 mM, native enzyme is activated to 111.6% of control, recombinant enzyme is activated to 109.1% of control, beta-glucosidase activity Putranjiva roxburghii
Fe2+ 1 mM, native enzyme is activated to 130.0% of control, recombinant enzyme is activated to 127.3% of control, beta-glucosidase activity Putranjiva roxburghii
Mn2+ 1 mM, native enzyme is activated to 102.3% of control, recombinant enzyme is activated to 106.4% of control, beta-glucosidase activity Putranjiva roxburghii
Pb2+ 1 mM, native enzyme is activated to 109.3% of control, recombinant enzyme is activated to 110.6% of control, beta-glucosidase activity Putranjiva roxburghii

Organism

Organism UniProt Comment Textmining
Putranjiva roxburghii A0A023MIF8
-
-

Purification (Commentary)

Purification (Comment) Organism
a single step concanvalin A affinity chromatography for native PRGH1 improves the yield and reduced the purification time Putranjiva roxburghii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-D-fucopyranoside + H2O relative activity of 125% as compared to 4-nitrophenyl beta-D-glucopyranoside. Maximum hydrolytic activity for beta-D-fucopyranoside followed by 4-nitrophenyl beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-fucopyranose
-
?
4-nitrophenyl beta-D-galactopyranoside + H2O maximum hydrolytic activity for beta-D-fucopyranoside followed by 4-nitrophenyl beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-galactopyranose
-
?
4-nitrophenyl beta-D-glucopyranoside + H2O maximum hydrolytic activity for beta-D-fucopyranoside followed by 4-nitrophenyl beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-glucopyranose
-
?
cellobiose + H2O
-
Putranjiva roxburghii 2 beta-D-glucose
-
?

Synonyms

Synonyms Comment Organism
PRGH1
-
Putranjiva roxburghii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
native and recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 65 native and recombinant enzymes are stable Putranjiva roxburghii
70
-
1 h, the native enzyme retains 67% of its activity while the recombinant enzyme preserves 35% Putranjiva roxburghii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.82
-
4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
6.95
-
cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
7.39
-
4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
8.12
-
cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
10.39
-
4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
12
-
4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
12.71
-
4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
14.77
-
4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.6
-
recombinant enzyme, glucosidase activity Putranjiva roxburghii

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 8 native and recombinant enzyme are stable over a broad pH range of 4.0-8.0 with maximum retention of their catalytic activity. However, activities decline quickly below pH 4.0 Putranjiva roxburghii

General Information

General Information Comment Organism
physiological function the enzyme may be involved in plant defence Putranjiva roxburghii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
7.99
-
cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
8.57
-
4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
10.1
-
cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
11.5
-
4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
14.9
-
4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
17.9
-
4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
22.6
-
4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
32.1
-
4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii