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Literature summary extracted from

  • Xia, W.; Bai, Y.; Cui, Y.; Xu, X.; Qian, L.; Shi, P.; Zhang, W.; Luo, H.; Zhan, X.; Yao, B.
    Functional diversity of family 3 beta-glucosidases from thermophilic cellulolytic fungus Humicola insolens Y1 (2016), Sci. Rep., 6, 27062.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.21 heterogeneous expression in Pichia pastoris strain GS115 Humicola insolens

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.21 F304I mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 I278F decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 I48W decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 I48W/I278F/T484Y decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 T484Y decreased activities towards 4-nitrophenyl beta-D-glucopyranoside, hydrolysis activity on sophorose, which is much higher than the activity on 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 W69I mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 Y509T introduction of mutation Y509T causes the increase of the Km value, but does not affect the kcat value Humicola insolens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.21 2-mercaptoethanol 5 mM, 22% inhibition Humicola insolens
3.2.1.21 Ag+ 5 mM, 72% inhibition; 5 mM, 76% inhibition; 5 mM, 89% inhibition Humicola insolens
3.2.1.21 Co2+ 5 mM, 10% inhibition Humicola insolens
3.2.1.21 Cr3+ 5 mM, 30% inhibition Humicola insolens
3.2.1.21 Cu2+ 5 mM, 6% inhibition Humicola insolens
3.2.1.21 D-glucose
-
 Humicola insolens
3.2.1.21 EDTA 5 mM, 9% inhibition Humicola insolens
3.2.1.21 Fe3+ 5 mM, 5% inhibition; 5 mM, 6% inhibition; 5 mM, 7% inhibition Humicola insolens
3.2.1.21 Mg2+ 5 mM, 23% loss of activity Humicola insolens
3.2.1.21 Ni2+ 5 mM, 10% inhibition Humicola insolens
3.2.1.21 Pb2+ 5 mM, 14% inhibition Humicola insolens
3.2.1.21 SDS 5 mM, 55% inhibition; 5 mM, 62% inhibition; 5 mM, 95% inhibition Humicola insolens
3.2.1.21 Zn2+ 5 mM, 10% loss of activity Humicola insolens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.21 0.2
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C Humicola insolens
3.2.1.21 0.9
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 1.51
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, wild-type enzyme Humicola insolens
3.2.1.21 1.99
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I48W Humicola insolens
3.2.1.21 2.37
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme T484Y Humicola insolens
3.2.1.21 2.42
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I278F Humicola insolens
3.2.1.21 2.45
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C, mutant enzyme Y509T Humicola insolens
3.2.1.21 2.48
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I48W/I278F/T484Y Humicola insolens
3.2.1.21 6.63
-
 cellobiose pH 5.5, 55°C Humicola insolens
3.2.1.21 8.44
-
 cellobiose pH 5.5, 60°C, wild-type enzyme Humicola insolens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.21 Mg2+ 5 mM, 1.1fold activiation Humicola insolens
3.2.1.21 Mn2+ 5 mM, 1.2fold activiation Humicola insolens
3.2.1.21 Mn2+ 5 mM, 1.3fold activiation Humicola insolens
3.2.1.21 additional information no remarkable activation or inhibition by 5 mM Mg2+, 5 mM Zn2+, 5 mM Ca2+, 5 mM Cu2+, 5 mM Co2+, 5 mM Cr2+, 5 mM Ni2+, 5 mM Pb2+, 5 mM EDTA, 5 mM 2-mercaptoethanol Humicola insolens
3.2.1.21 Zn2+ 5 mM, 1.1fold activiation Humicola insolens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.21 78280
-
 calculated from sequence Humicola insolens
3.2.1.21 80000
-
-
 Humicola insolens
3.2.1.21 96870
-
 calculated from sequence Humicola insolens
3.2.1.21 97120
-
 calculated from sequence Humicola insolens
3.2.1.21 120000
-
 1 * 120000, SDS-PAGE Humicola insolens
3.2.1.21 127000
-
-
 Humicola insolens

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.21 Humicola insolens A0A191KCR0
-
-
3.2.1.21 Humicola insolens A0A191KCR1
-
-
3.2.1.21 Humicola insolens A0A191KCR2
-
-
3.2.1.21 Humicola insolens Y1 A0A191KCR0
-
-
3.2.1.21 Humicola insolens Y1 A0A191KCR1
-
-
3.2.1.21 Humicola insolens Y1 A0A191KCR2
-
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.2.1.21 glycoprotein 12 putative N-glycosylation sites Humicola insolens
3.2.1.21 glycoprotein 2 putative N-glycosylation sites Humicola insolens
3.2.1.21 glycoprotein 7 putative N-glycosylation sites Humicola insolens

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.21
-
 Humicola insolens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.21 0.95
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 6.0, 50°C, mutant enzyme I48W Humicola insolens
3.2.1.21 1
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 6.0, 50°C, mutant enzyme I48W/I278F/T484Y Humicola insolens
3.2.1.21 2.68
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 6.0, 50°C, mutant enzyme I278F Humicola insolens
3.2.1.21 3.36
-
 substrate: sophorose, pH 6.0, 50°C, mutant enzyme I48W/I278F/T484Y Humicola insolens
3.2.1.21 3.59
-
 substrate: sophorose, pH 6.0, 50°C, mutant enzyme I278F Humicola insolens
3.2.1.21 3.7
-
 substrate: sophorose, pH 6.0, 50°C, mutant enzyme I48W Humicola insolens
3.2.1.21 4.46
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 6.0, 50°C, mutant enzyme T484Y Humicola insolens
3.2.1.21 5.64
-
 substrate: sophorose, pH 6.0, 50°C, mutant enzyme T484Y Humicola insolens
3.2.1.21 22.3
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 5.5, 60°C, mutant enzyme Y509T Humicola insolens
3.2.1.21 31.6
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 6.0, 50°C, wild-type enzyme Humicola insolens
3.2.1.21 36.3
-
 substrate: substrate: cellobiose, pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 56.4
-
 substrate: cellobiose, pH 5.5, 55°C Humicola insolens
3.2.1.21 57.5
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 62.6
-
 substrate: daidzin, pH 6.0, 50°C Humicola insolens
3.2.1.21 89
-
 substrate: daidzin, pH 5.5, 55°C Humicola insolens
3.2.1.21 94.6
-
 substrate: sophorose, pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 103.4
-
 substrate: gentiobiose, pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 113.9
-
 substrate: sophorose, pH 5.5, 55°C Humicola insolens
3.2.1.21 158.8
-
 substrate: 4-nitrophenyl beta-D-glucopyranoside, pH 5.5, 60°C Humicola insolens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.21 4-nitrophenyl alpha-L-arabinofuranoside + H2O
-
 Humicola insolens 4-nitrophenol + alpha-L-arabinofuranose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-cellobioside + H2O
-
 Humicola insolens 4-nitrophenol + cellobiose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-cellobioside + H2O
-
 Humicola insolens Y1 4-nitrophenol + cellobiose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-galactoside + H2O
-
 Humicola insolens 4-nitrophenol + beta-D-galactose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-galactoside + H2O
-
 Humicola insolens Y1 4-nitrophenol + beta-D-galactose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Humicola insolens 4-nitrophenol + D-glucopyranose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Humicola insolens Y1 4-nitrophenol + D-glucopyranose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-xylopyranoside + H2O
-
 Humicola insolens 4-nitrophenol + beta-D-xylopyranose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-xylopyranoside + H2O
-
 Humicola insolens Y1 4-nitrophenol + beta-D-xylopyranose
-
 ?
3.2.1.21 cellobiose + H2O
-
 Humicola insolens 2 D-glucose
-
 ?
3.2.1.21 cellobiose + H2O
-
 Humicola insolens Y1 2 D-glucose
-
 ?
3.2.1.21 daidzin + H2O
-
 Humicola insolens beta-D-glucose + daidzein
-
 ?
3.2.1.21 daidzin + H2O
-
 Humicola insolens Y1 beta-D-glucose + daidzein
-
 ?
3.2.1.21 genistin + H2O
-
 Humicola insolens beta-D-glucose + genistein
-
 ?
3.2.1.21 gentiobiose + H2O
-
 Humicola insolens D-glucose
-
 ?
3.2.1.21 gentiobiose + H2O
-
 Humicola insolens Y1 D-glucose
-
 ?
3.2.1.21 glycitin + H2O
-
 Humicola insolens beta-D-glucose + glycitein
-
 ?
3.2.1.21 sophorose + H2O
-
 Humicola insolens 2 D-glucose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.21 monomer 1 * 120000, SDS-PAGE Humicola insolens
3.2.1.21 monomer 1 * 127000, calculated from sequence Humicola insolens
3.2.1.21 monomer 1 * 78276, calculated from sequence Humicola insolens
3.2.1.21 monomer 1 * 80000, calculated from sequence Humicola insolens
3.2.1.21 monomer 1 * 96873, calculated from sequence Humicola insolens
3.2.1.21 monomer 1 * 97117, calculated from sequence Humicola insolens

Synonyms

EC Number Synonyms Comment Organism
3.2.1.21 HiBgl3A
-
 Humicola insolens
3.2.1.21 HiBgl3B
-
 Humicola insolens
3.2.1.21 HiBgl3C
-
 Humicola insolens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.21 50 55 with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens
3.2.1.21 55
-
 with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens
3.2.1.21 60
-
 with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.21 40 60 40°C: about 50% of maximal activity, 60°C: about 85% of maximal activity, substrate: 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 40 60 40°C: about 65% of maximal activity, 60°C: about 70% of maximal activity, substrate: 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 50 65 50°C: about 45% of maximal activity, 65°C: about 50% of maximal activity, substrate: 4-nitrophenyl beta-D-glucopyranoside Humicola insolens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.21 50
-
 1 h, no loss of activity Humicola insolens
3.2.1.21 50
-
 1 h, about 10% loss of activity Humicola insolens
3.2.1.21 60
-
 1 h, about 55% loss of activity Humicola insolens
3.2.1.21 60
-
 30 min, about 90% loss of activity Humicola insolens
3.2.1.21 70
-
 1 h, 10 min, 95% loss of activity Humicola insolens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.21 3.21
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I48W Humicola insolens
3.2.1.21 4.24
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I48W/I278F/T484Y Humicola insolens
3.2.1.21 9.61
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I278F Humicola insolens
3.2.1.21 15.05
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme T484Y Humicola insolens
3.2.1.21 40.42
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, wild-type enzyme Humicola insolens
3.2.1.21 68.16
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C, mutant enzyme Y509T Humicola insolens
3.2.1.21 73.08
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C, wild-type enzyme Humicola insolens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.21 5 5.5 with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens
3.2.1.21 5.5
-
 with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens
3.2.1.21 6
-
 with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.21 4 8 pH 4.0: about 30% of maximal activity, pH 8.0: about 30% of maximal activity, substrate: 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 5 6.5 pH 5.0: about 30% of maximal activity, pH 6.5: about 60% of maximal activity, substrate: 4-nitrophenyl beta-D-glucopyranoside Humicola insolens
3.2.1.21 5 6.5 pH 5.0: about 90% of maximal activity, pH 6.5: about 70% of maximal activity, substrate: 4-nitrophenyl beta-D-glucopyranoside Humicola insolens

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.21 5 9 37°C, 1 h, enzyme retains more than 80% activity Humicola insolens
3.2.1.21 5 10 37°C, 1 h, enzyme retains more than 80% activity Humicola insolens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.21 2 5 D-glucose pH 5.5, 60°C, with 4-nitrophenyl beta-D-glucopyranoside as substrate, wild-type enzyme Humicola insolens
3.2.1.21 37.1
-
 D-glucose pH 5.5, 55°C, with 4-nitrophenyl beta-D-glucopyranoside as substrate Humicola insolens
3.2.1.21 55.2
-
 D-glucose pH 6.0, 50°C, with 4-nitrophenyl beta-D-glucopyranoside as substrat, wild-type enzyme Humicola insolens

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.2.1.21 Humicola insolens calculated
-
 5.93
3.2.1.21 Humicola insolens calculated
-
 6.03
3.2.1.21 Humicola insolens calculated
-
 6.26

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 1.61
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I48W Humicola insolens
3.2.1.21 1.71
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I48W/I278F/T484Y Humicola insolens
3.2.1.21 3.98
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme I278F Humicola insolens
3.2.1.21 6.36
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, mutant enzyme T484Y Humicola insolens
3.2.1.21 11.1
-
 cellobiose pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 23
-
 cellobiose pH 5.5, 55°C Humicola insolens
3.2.1.21 26.77
-
 4-nitrophenyl beta-D-glucopyranoside pH 6.0, 50°C, wild-type enzyme Humicola insolens
3.2.1.21 27.82
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C, mutant enzyme Y509T Humicola insolens
3.2.1.21 81.6
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C, wild-type enzyme Humicola insolens
3.2.1.21 1557
-
 4-nitrophenyl beta-D-glucopyranoside pH 5.5, 60°C Humicola insolens