Literature summary for 3.2.1.21 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.

Search for more literature for 3.2.1.21

Cloned(Commentary)

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

Protein Variants

Protein Variants	Comment	Organism
F304I	mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside	Humicola insolens
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
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
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
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
W69I	mutant enzyme shows no activity with 4-nitrophenyl beta-D-glucopyranoside	Humicola insolens
Y509T	introduction of mutation Y509T causes the increase of the Km value, but does not affect the kcat value	Humicola insolens

Inhibitors

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

KM Value [mM]

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

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	5 mM, 1.1fold activiation	Humicola insolens
Mn2+	5 mM, 1.2fold activiation	Humicola insolens
Mn2+	5 mM, 1.3fold activiation	Humicola insolens
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
Zn2+	5 mM, 1.1fold activiation	Humicola insolens

Molecular Weight [Da]

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

Organism

Organism	UniProt	Comment	Textmining
Humicola insolens	A0A191KCR0	-	-
Humicola insolens	A0A191KCR1	-	-
Humicola insolens	A0A191KCR2	-	-
Humicola insolens Y1	A0A191KCR0	-	-
Humicola insolens Y1	A0A191KCR1	-	-
Humicola insolens Y1	A0A191KCR2	-	-

Posttranslational Modification

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

Purification (Commentary)

Purification (Comment)	Organism
-	Humicola insolens

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
HiBgl3A	-	Humicola insolens
HiBgl3B	-	Humicola insolens
HiBgl3C	-	Humicola insolens

Temperature Optimum [°C]

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

Temperature Range [°C]

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

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

Turnover Number [1/s]

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

pH Optimum

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

pH Range

pH Minimum	pH Maximum	Comment	Organism
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
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
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

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

Ki Value [mM]

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

pI Value

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

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
1.61	-	4-nitrophenyl beta-D-glucopyranoside	pH 6.0, 50°C, mutant enzyme I48W	Humicola insolens
1.71	-	4-nitrophenyl beta-D-glucopyranoside	pH 6.0, 50°C, mutant enzyme I48W/I278F/T484Y	Humicola insolens
3.98	-	4-nitrophenyl beta-D-glucopyranoside	pH 6.0, 50°C, mutant enzyme I278F	Humicola insolens
6.36	-	4-nitrophenyl beta-D-glucopyranoside	pH 6.0, 50°C, mutant enzyme T484Y	Humicola insolens
11.1	-	cellobiose	pH 5.5, 60°C, wild-type enzyme	Humicola insolens
23	-	cellobiose	pH 5.5, 55°C	Humicola insolens
26.77	-	4-nitrophenyl beta-D-glucopyranoside	pH 6.0, 50°C, wild-type enzyme	Humicola insolens
27.82	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.5, 60°C, mutant enzyme Y509T	Humicola insolens
81.6	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.5, 60°C, wild-type enzyme	Humicola insolens
1557	-	4-nitrophenyl beta-D-glucopyranoside	pH 5.5, 60°C	Humicola insolens

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Release 2023.1 (January 2023)