Literature summary for 3.2.1.21 extracted from

Kulkarni, T.; Khan, S.; Villagomez, R.; Mahmood, T.; Lindahl, S.; Logan, D.; Linares-Pasten, J.; Nordberg Karlsson, E.
Crystal structure of beta-glucosidase 1A from Thermotoga neapolitana and comparison of active site mutants for hydrolysis of flavonoid glucosides (2017), Proteins, 85, 872-884 .

Search for more literature for 3.2.1.21

Crystallization (Commentary)

Crystallization (Comment)	Organism
mutant E349G, to 1.9 A resolution, and docking studies of flavonoid substrates	Thermotoga neapolitana

Protein Variants

Protein Variants	Comment	Organism
E349G	crystallization data	Thermotoga neapolitana
F224I	mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected	Thermotoga neapolitana
F310E	mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected	Thermotoga neapolitana
F310L	mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected	Thermotoga neapolitana
N220F	mutation the aglycone binding region	Thermotoga neapolitana
N220S	mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside	Thermotoga neapolitana
N221F	mutation the aglycone binding region	Thermotoga neapolitana
N221S	mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside	Thermotoga neapolitana
W322A	mutation the aglycone binding region, residue is important for substrate accomodation. Mutation W322A results in a large reduction of hydrolytic activity on 3-glucosylated flavonoids	Thermotoga neapolitana

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.09	-	quercetin 3-O-beta-D-glucopyranoside	mutant N220S, pH 5.6, 80°C	Thermotoga neapolitana
0.09	-	quercetin 3-O-beta-D-glucopyranoside	mutant N221S, pH 5.6, 80°C	Thermotoga neapolitana
0.13	-	quercetin 3-O-beta-D-glucopyranoside	wild-type, pH 5.6, 80°C	Thermotoga neapolitana
0.15	-	quercetin 3-O-beta-D-glucopyranoside	mutant F224I, pH 5.6, 80°C	Thermotoga neapolitana
0.17	-	4-nitrophenyl beta-D-glucopyranoside	mutant F310L, pH 5.6, 80°C	Thermotoga neapolitana
0.19	-	4-nitrophenyl beta-D-glucopyranoside	mutant F310E, pH 5.6, 80°C	Thermotoga neapolitana
0.22	-	4-nitrophenyl beta-D-glucopyranoside	mutant W322A, pH 5.6, 80°C	Thermotoga neapolitana
0.24	-	4-nitrophenyl beta-D-glucopyranoside	mutant N221F, pH 5.6, 80°C	Thermotoga neapolitana
0.24	-	4-nitrophenyl beta-D-glucopyranoside	wild-type, pH 5.6, 80°C	Thermotoga neapolitana
0.3	-	4-nitrophenyl beta-D-glucopyranoside	mutant N220F, pH 5.6, 80°C	Thermotoga neapolitana
0.31	-	4-nitrophenyl beta-D-glucopyranoside	mutant N220S, pH 5.6, 80°C	Thermotoga neapolitana
0.43	-	4-nitrophenyl beta-D-glucopyranoside	mutant N221S, pH 5.6, 80°C	Thermotoga neapolitana
0.64	-	4-nitrophenyl beta-D-glucopyranoside	mutant F224I, pH 5.6, 80°C	Thermotoga neapolitana

Organism

Organism	UniProt	Comment	Textmining
Thermotoga neapolitana	B9K7M5	cf. EC 3.2.1.74	-
Thermotoga neapolitana DSM 4359	B9K7M5	cf. EC 3.2.1.74	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl beta-D-glucopyranoside + H2O	-	Thermotoga neapolitana	4-nitrophenol + beta-D-glucose	-	?
4-nitrophenyl beta-D-glucopyranoside + H2O	-	Thermotoga neapolitana DSM 4359	4-nitrophenol + beta-D-glucose	-	?
daidzein-7-O-beta-D-glucopyranoside + H2O	-	Thermotoga neapolitana	daidzein + D-glucose	-	?
daidzein-7-O-beta-D-glucopyranoside + H2O	-	Thermotoga neapolitana DSM 4359	daidzein + D-glucose	-	?
quercetin 3,4'-O-di-beta-D-glucopyranoside + 2 H2O	-	Thermotoga neapolitana	quercetin + 2 D-glucose	-	?
quercetin 3,4'-O-di-beta-D-glucopyranoside + 2 H2O	-	Thermotoga neapolitana DSM 4359	quercetin + 2 D-glucose	-	?
quercetin 3-O-beta-D-glucopyranoside + H2O	-	Thermotoga neapolitana	quercetin + D-glucose	-	?
quercetin 3-O-beta-D-glucopyranoside + H2O	-	Thermotoga neapolitana DSM 4359	quercetin + D-glucose	-	?

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
80	-	assay at	Thermotoga neapolitana

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
100.3	-	mutant W322A, melting temperature	Thermotoga neapolitana
100.8	-	mutant N221S, melting temperature	Thermotoga neapolitana
101.1	-	mutant F224I, melting temperature	Thermotoga neapolitana
101.4	-	mutant F310L, melting temperature	Thermotoga neapolitana
101.6	-	mutant N220S, melting temperature	Thermotoga neapolitana
101.9	-	wild-type, melting temperature	Thermotoga neapolitana

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
13.6	-	quercetin 3-O-beta-D-glucopyranoside	wild-type, pH 5.6, 80°C	Thermotoga neapolitana
14.4	-	quercetin 3-O-beta-D-glucopyranoside	mutant F224I, pH 5.6, 80°C	Thermotoga neapolitana
22	-	4-nitrophenyl beta-D-glucopyranoside	mutant N221F, pH 5.6, 80°C	Thermotoga neapolitana
39.8	-	quercetin 3-O-beta-D-glucopyranoside	mutant N220S, pH 5.6, 80°C	Thermotoga neapolitana
40.8	-	quercetin 3-O-beta-D-glucopyranoside	mutant N221S, pH 5.6, 80°C	Thermotoga neapolitana
61	-	4-nitrophenyl beta-D-glucopyranoside	mutant W322A, pH 5.6, 80°C	Thermotoga neapolitana
371	-	4-nitrophenyl beta-D-glucopyranoside	mutant F310L, pH 5.6, 80°C	Thermotoga neapolitana
485	-	4-nitrophenyl beta-D-glucopyranoside	wild-type, pH 5.6, 80°C	Thermotoga neapolitana
508	-	4-nitrophenyl beta-D-glucopyranoside	mutant F310E, pH 5.6, 80°C	Thermotoga neapolitana
757	-	4-nitrophenyl beta-D-glucopyranoside	mutant N220S, pH 5.6, 80°C	Thermotoga neapolitana
803	-	4-nitrophenyl beta-D-glucopyranoside	mutant F224I, pH 5.6, 80°C	Thermotoga neapolitana
849	-	4-nitrophenyl beta-D-glucopyranoside	mutant N220F, pH 5.6, 80°C	Thermotoga neapolitana
1170	-	4-nitrophenyl beta-D-glucopyranoside	mutant N221S, pH 5.6, 80°C	Thermotoga neapolitana

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5.6	-	assay at	Thermotoga neapolitana

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
93	-	4-nitrophenyl beta-D-glucopyranoside	mutant N221F, pH 5.6, 80°C	Thermotoga neapolitana
95	-	quercetin 3-O-beta-D-glucopyranoside	mutant F224I, pH 5.6, 80°C	Thermotoga neapolitana
101	-	quercetin 3-O-beta-D-glucopyranoside	wild-type, pH 5.6, 80°C	Thermotoga neapolitana
282	-	4-nitrophenyl beta-D-glucopyranoside	mutant W322A, pH 5.6, 80°C	Thermotoga neapolitana
428	-	quercetin 3-O-beta-D-glucopyranoside	mutant N220S, pH 5.6, 80°C	Thermotoga neapolitana
446	-	quercetin 3-O-beta-D-glucopyranoside	mutant N221S, pH 5.6, 80°C	Thermotoga neapolitana
1247	-	4-nitrophenyl beta-D-glucopyranoside	mutant F224I, pH 5.6, 80°C	Thermotoga neapolitana
2000	-	4-nitrophenyl beta-D-glucopyranoside	wild-type, pH 5.6, 80°C	Thermotoga neapolitana
2144	-	4-nitrophenyl beta-D-glucopyranoside	mutant F310L, pH 5.6, 80°C	Thermotoga neapolitana
2433	-	4-nitrophenyl beta-D-glucopyranoside	mutant N220S, pH 5.6, 80°C	Thermotoga neapolitana
2714	-	4-nitrophenyl beta-D-glucopyranoside	mutant F310E, pH 5.6, 80°C	Thermotoga neapolitana
2724	-	4-nitrophenyl beta-D-glucopyranoside	mutant N221S, pH 5.6, 80°C	Thermotoga neapolitana
2857	-	4-nitrophenyl beta-D-glucopyranoside	mutant N220F, pH 5.6, 80°C	Thermotoga neapolitana

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Release 2023.1 (January 2023)