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Literature summary 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 .
    View publication on PubMed

Crystallization (Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.21 E349G crystallization data Thermotoga neapolitana
3.2.1.21 F224I mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected Thermotoga neapolitana
3.2.1.21 F310E mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected Thermotoga neapolitana
3.2.1.21 F310L mutation the aglycone binding region, flavonoid glucoside hydrolysis is not affected Thermotoga neapolitana
3.2.1.21 N220F mutation the aglycone binding region Thermotoga neapolitana
3.2.1.21 N220S mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside Thermotoga neapolitana
3.2.1.21 N221F mutation the aglycone binding region Thermotoga neapolitana
3.2.1.21 N221S mutation the aglycone binding region, increases the catalytic efficiency on both quercetin 3-glucoside and 4-nitrophenyl-beta-D-glucoside Thermotoga neapolitana
3.2.1.21 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.21 0.09
-
 quercetin 3-O-beta-D-glucopyranoside mutant N220S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.09
-
 quercetin 3-O-beta-D-glucopyranoside mutant N221S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.13
-
 quercetin 3-O-beta-D-glucopyranoside wild-type, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.15
-
 quercetin 3-O-beta-D-glucopyranoside mutant F224I, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.17
-
 4-nitrophenyl beta-D-glucopyranoside mutant F310L, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.19
-
 4-nitrophenyl beta-D-glucopyranoside mutant F310E, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.22
-
 4-nitrophenyl beta-D-glucopyranoside mutant W322A, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.24
-
 4-nitrophenyl beta-D-glucopyranoside mutant N221F, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.24
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.3
-
 4-nitrophenyl beta-D-glucopyranoside mutant N220F, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.31
-
 4-nitrophenyl beta-D-glucopyranoside mutant N220S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.43
-
 4-nitrophenyl beta-D-glucopyranoside mutant N221S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 0.64
-
 4-nitrophenyl beta-D-glucopyranoside mutant F224I, pH 5.6, 80°C Thermotoga neapolitana

Organism

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.21 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermotoga neapolitana 4-nitrophenol + beta-D-glucose
-
 ?
3.2.1.21 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermotoga neapolitana DSM 4359 4-nitrophenol + beta-D-glucose
-
 ?
3.2.1.21 daidzein-7-O-beta-D-glucopyranoside + H2O
-
 Thermotoga neapolitana daidzein + D-glucose
-
 ?
3.2.1.21 daidzein-7-O-beta-D-glucopyranoside + H2O
-
 Thermotoga neapolitana DSM 4359 daidzein + D-glucose
-
 ?
3.2.1.21 quercetin 3,4'-O-di-beta-D-glucopyranoside + 2 H2O
-
 Thermotoga neapolitana quercetin + 2 D-glucose
-
 ?
3.2.1.21 quercetin 3,4'-O-di-beta-D-glucopyranoside + 2 H2O
-
 Thermotoga neapolitana DSM 4359 quercetin + 2 D-glucose
-
 ?
3.2.1.21 quercetin 3-O-beta-D-glucopyranoside + H2O
-
 Thermotoga neapolitana quercetin + D-glucose
-
 ?
3.2.1.21 quercetin 3-O-beta-D-glucopyranoside + H2O
-
 Thermotoga neapolitana DSM 4359 quercetin + D-glucose
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.21 80
-
 assay at Thermotoga neapolitana

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.21 100.3
-
 mutant W322A, melting temperature Thermotoga neapolitana
3.2.1.21 100.8
-
 mutant N221S, melting temperature Thermotoga neapolitana
3.2.1.21 101.1
-
 mutant F224I, melting temperature Thermotoga neapolitana
3.2.1.21 101.4
-
 mutant F310L, melting temperature Thermotoga neapolitana
3.2.1.21 101.6
-
 mutant N220S, melting temperature Thermotoga neapolitana
3.2.1.21 101.9
-
 wild-type, melting temperature Thermotoga neapolitana

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.21 13.6
-
 quercetin 3-O-beta-D-glucopyranoside wild-type, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 14.4
-
 quercetin 3-O-beta-D-glucopyranoside mutant F224I, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 22
-
 4-nitrophenyl beta-D-glucopyranoside mutant N221F, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 39.8
-
 quercetin 3-O-beta-D-glucopyranoside mutant N220S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 40.8
-
 quercetin 3-O-beta-D-glucopyranoside mutant N221S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 61
-
 4-nitrophenyl beta-D-glucopyranoside mutant W322A, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 371
-
 4-nitrophenyl beta-D-glucopyranoside mutant F310L, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 485
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 508
-
 4-nitrophenyl beta-D-glucopyranoside mutant F310E, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 757
-
 4-nitrophenyl beta-D-glucopyranoside mutant N220S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 803
-
 4-nitrophenyl beta-D-glucopyranoside mutant F224I, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 849
-
 4-nitrophenyl beta-D-glucopyranoside mutant N220F, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 1170
-
 4-nitrophenyl beta-D-glucopyranoside mutant N221S, pH 5.6, 80°C Thermotoga neapolitana

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.21 5.6
-
 assay at Thermotoga neapolitana

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 93
-
 4-nitrophenyl beta-D-glucopyranoside mutant N221F, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 95
-
 quercetin 3-O-beta-D-glucopyranoside mutant F224I, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 101
-
 quercetin 3-O-beta-D-glucopyranoside wild-type, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 282
-
 4-nitrophenyl beta-D-glucopyranoside mutant W322A, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 428
-
 quercetin 3-O-beta-D-glucopyranoside mutant N220S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 446
-
 quercetin 3-O-beta-D-glucopyranoside mutant N221S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 1247
-
 4-nitrophenyl beta-D-glucopyranoside mutant F224I, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 2000
-
 4-nitrophenyl beta-D-glucopyranoside wild-type, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 2144
-
 4-nitrophenyl beta-D-glucopyranoside mutant F310L, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 2433
-
 4-nitrophenyl beta-D-glucopyranoside mutant N220S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 2714
-
 4-nitrophenyl beta-D-glucopyranoside mutant F310E, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 2724
-
 4-nitrophenyl beta-D-glucopyranoside mutant N221S, pH 5.6, 80°C Thermotoga neapolitana
3.2.1.21 2857
-
 4-nitrophenyl beta-D-glucopyranoside mutant N220F, pH 5.6, 80°C Thermotoga neapolitana