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Literature summary for 3.2.1.21 extracted from

  • Nakabayashi, M.; Kataoka, M.; Mishima, Y.; Maeno, Y.; Ishikawa, K.
    Structural analysis of beta-glucosidase mutants derived from a hyperthermophilic tetrameric structure (2014), Acta Crystallogr. Sect. D, 70, 877-888.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 Pyrococcus furiosus

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme R170A/R220A/Y227F, hanging-drop vapour-diffusion method, the structure is determined at 2.35 A resolution, stable homotetrameric structure, a dimeric form of the enzyme is constructed and its crystal structure is solved at a resolution of 1.70 A Pyrococcus furiosus

Protein Variants

Protein Variants Comment Organism
R170A/R220A/Y227F mutant enzyme occurs as dimer Pyrococcus furiosus
R170A/R220A/Y227F/R448E mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme Pyrococcus furiosus
R170A/R220A/Y227F/R449R mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme Pyrococcus furiosus
R170A/R220A/Y227F/R459G mutant enzyme occurs as monomer, melting temperature is decreased form 110°C for the wild-type enzyme to 73-75°C for the mutant enzyme Pyrococcus furiosus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23600
-
wild-type enzyme tetramer, gel filtration Pyrococcus furiosus
50000
-
mutant enzyme R170A/R220A/Y227F/R459G, monomer, gel filtration Pyrococcus furiosus
55000
-
2 * 55000, SDS-PAGE after heating at 95°C,mutant enzyme R170A/R220A/Y227F Pyrococcus furiosus
71000
-
mutant enzyme R170A/R220A/Y227F/R448E, monomer, gel filtration Pyrococcus furiosus
96000
-
mutant enzyme R170A/R220A/Y227F/R449R, monomer, gel filtration Pyrococcus furiosus
136000
-
mutant enzyme R170A/R220A/Y227F, dimer, gel filtration Pyrococcus furiosus

Organism

Organism UniProt Comment Textmining
Pyrococcus furiosus Q51723
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzyme R170A/R220A/Y227F Pyrococcus furiosus

Subunits

Subunits Comment Organism
dimer 2 * 55000, SDS-PAGE after heating at 95°C,mutant enzyme R170A/R220A/Y227F Pyrococcus furiosus
monomer 1 * 50000, mutant enzyme R170A/R220A/Y227F/R459G, SDS-PAGE Pyrococcus furiosus
monomer 1 * 71000, mutant enzyme R170A/R220A/Y227F/R448E, SDS-PAGE Pyrococcus furiosus
monomer 1 * 96000, mutant enzyme R170A/R220A/Y227F/R449R, SDS-PAGE Pyrococcus furiosus
tetramer wild-type enzyme Pyrococcus furiosus

Synonyms

Synonyms Comment Organism
BGLPf
-
Pyrococcus furiosus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
oligomerization contributes to thermostability Pyrococcus furiosus
70 80 mutant enzymes R170A/R220A/Y227F/R448E, R170A/R220A/Y227F/R449R or R170A/R220A/Y227F/R459G are immediately inactivated Pyrococcus furiosus
73 75 melting temperature of mutant enzymes R170A/R220A/Y227F/R448E, R170A/R220A/Y227F/R449R or R170A/R220A/Y227F/R459G Pyrococcus furiosus
102
-
Tm-value of mutant enzyme R170A/R220A/Y227F Pyrococcus furiosus
110
-
melting temperature of tetrameric wild-type enzyme Pyrococcus furiosus