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Literature summary 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

EC Number Activating Compound Comment Organism Structure
3.2.1.21 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
3.2.1.21 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
3.2.1.21 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
3.2.1.21 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

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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.21 overexpression in Escherichia coli Putranjiva roxburghii
3.2.1.23 overexpression in Escherichia coli Putranjiva roxburghii
3.2.1.38 overexpression in Escherichia coli Putranjiva roxburghii

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.21 E173N mutant enzyme shows no activity Putranjiva roxburghii
3.2.1.21 E389N mutant enzyme shows no activity Putranjiva roxburghii
3.2.1.21 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
3.2.1.21 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
3.2.1.23 E173N mutant enzyme shows no activity Putranjiva roxburghii
3.2.1.23 E389N mutant enzyme shows no activity Putranjiva roxburghii
3.2.1.23 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
3.2.1.23 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
3.2.1.38 E173N mutant enzyme shows no activity Putranjiva roxburghii
3.2.1.38 E389N mutant enzyme shows no activity Putranjiva roxburghii
3.2.1.38 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
3.2.1.38 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

EC Number Inhibitors Comment Organism Structure
3.2.1.21 2-mercaptoethanol 1 mM, 9% loss of activity of native enzyme, 10% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 Cr3+ 1 mM, 29% loss of activity of native enzyme, 30% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 delta-gluconolactone 5 mM, 85% loss of activity of native enzyme, 86% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 DTT 1 mM, 4% loss of activity of native enzyme, 5% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 Guanidine HCl 10 mM, 8% loss of activity of native enzyme, 10% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 Hg2+ 1 mM, 55% loss of activity of native enzyme, 58% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 Mg2+ 1 mM, 7% loss of activity, native and recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 additional information significant resistant to D-glucose concentration up to 500 mM Putranjiva roxburghii
3.2.1.21 N-bromosuccinamide 2 mM, 92% loss of activity of native enzyme, 94% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 Ni2+ 1 mM, 16% loss of activity of native enzyme, 18% loss of activity of recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii
3.2.1.21 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.21 0.46
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 0.51
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 0.53
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 0.58
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 0.64
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 0.68
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 0.81
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 0.87
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 0.46
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 0.51
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 0.53
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 0.58
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 0.64
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 0.68
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 0.81
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 0.87
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 0.46
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 0.51
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 0.53
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 0.58
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 0.64
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 0.68
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 0.81
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 0.87
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.21 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
3.2.1.21 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
3.2.1.21 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
3.2.1.21 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

EC Number Organism UniProt Comment Textmining
3.2.1.21 Putranjiva roxburghii A0A023MIF8
-
-
3.2.1.23 Putranjiva roxburghii A0A023MIF8
-
-
3.2.1.38 Putranjiva roxburghii A0A023MIF8
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.21 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
-
 ?
3.2.1.21 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
-
 ?
3.2.1.21 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
-
 ?
3.2.1.21 cellobiose + H2O
-
 Putranjiva roxburghii 2 beta-D-glucose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-fucopyranoside + H2O relative activity of 125% as compared to 4-nitrophenyl beta-D-glucopyranoside. Maximum hydrolytic activity for 4-nitrophenyl beta-D-fucopyranoside followed by 4-nitrophenyl beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-fucopyranose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-galactopyranoside + H2O maximum hydrolytic activity for 4-nitrophenyl beta-D-fucopyranoside followed by 4-nitrophenyl beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-galactopyranose
-
 ?
3.2.1.23 4-nitrophenyl beta-D-glucopyranoside + H2O maximum hydrolytic activity for 4-nitrophenyl beta-D-fucopyranoside followed by 4-nitrophenyl beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-glucopyranose
-
 ?
3.2.1.23 cellobiose + H2O
-
 Putranjiva roxburghii 2 beta-D-glucose
-
 ?
3.2.1.38 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 beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-fucopyranose
-
 ?
3.2.1.38 4-nitrophenyl beta-D-galactopyranoside + H2O maximum hydrolytic activity for beta-D-fucopyranoside followed by beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-galactopyranose
-
 ?
3.2.1.38 4-nitrophenyl beta-D-glucopyranoside + H2O maximum hydrolytic activity for beta-D-fucopyranoside followed by beta-D-glucopyranoside and 4-nitrophenyl beta-D-galactopyranoside Putranjiva roxburghii 4-nitrophenol + beta-D-glucopyranose
-
 ?
3.2.1.38 cellobiose + H2O
-
 Putranjiva roxburghii 2 beta-D-glucose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.21 PRGH1
-
 Putranjiva roxburghii
3.2.1.23 PRGH1
-
 Putranjiva roxburghii
3.2.1.38 PRGH1
-
 Putranjiva roxburghii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.21 65
-
 native and recombinant enzyme, beta-glucosidase activity Putranjiva roxburghii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.21 25 65 native and recombinant enzymes are stable Putranjiva roxburghii
3.2.1.21 70
-
 1 h, the native enzyme retains 67% of its activity while the recombinant enzyme preserves 35% Putranjiva roxburghii
3.2.1.23 25 65 native and recombinant enzyme are stable Putranjiva roxburghii
3.2.1.23 70
-
 1 h, the native enzyme retains 67% of its activity while the recombinant enzyme preserves 35% Putranjiva roxburghii
3.2.1.38 25 65 native and recombinant enzyme are stable Putranjiva roxburghii
3.2.1.38 70
-
 1 h, the native enzyme retains 67% of its activity while the recombinant enzyme preserves 35% Putranjiva roxburghii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.21 5.82
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 6.95
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 7.39
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 8.12
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 10.39
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 12
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 12.71
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 14.77
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 5.82
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 6.95
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 7.39
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 8.12
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 10.39
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 12
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 12.71
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 14.77
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.21 4.6
-
 recombinant enzyme, glucosidase activity Putranjiva roxburghii
3.2.1.23 4.4
-
 recombinant enzyme, galactosidase activity Putranjiva roxburghii
3.2.1.38 4.6
-
 native and recombinant enzyme, fucosidase activity Putranjiva roxburghii

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.21 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
3.2.1.23 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
3.2.1.38 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

EC Number General Information Comment Organism
3.2.1.21 physiological function the enzyme may be involved in plant defence Putranjiva roxburghii
3.2.1.23 physiological function the enzyme may be involved in plant defence Putranjiva roxburghii
3.2.1.38 physiological function the enzyme may be involved in plant defence Putranjiva roxburghii

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.21 7.99
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 8.57
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 10.1
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 11.5
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 14.9
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 17.9
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.21 22.6
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.21 32.1
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 7.99
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 8.57
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 10.1
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 11.5
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 14.9
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 17.9
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.23 22.6
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.23 32.1
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 7.99
-
 cellobiose pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 8.57
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 10.1
-
 cellobiose pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 11.5
-
 4-nitrophenyl beta-D-galactopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 14.9
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 17.9
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, recombinant enzyme Putranjiva roxburghii
3.2.1.38 22.6
-
 4-nitrophenyl beta-D-glucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii
3.2.1.38 32.1
-
 4-nitrophenyl beta-D-fucopyranoside pH 4.6, 37°C, native enzyme Putranjiva roxburghii