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Literature summary for 3.2.1.B26 extracted from

  • Shin, K.C.; Choi, H.Y.; Seo, M.J.; Oh, D.K.
    Improved conversion of ginsenoside Rb to compound K by semi-rational design of Sulfolobus solfataricus beta-glycosidase (2017), AMB Express, 7, 186 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.B26

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli strain ER2566 Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
F222A the mutant shows 105% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
F359A the mutant shows 84% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
L213A the mutant shows 110% activity with ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
S220A the mutant shows 92% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
V209A the mutant shows 170% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
V209D inactive Saccharolobus solfataricus
V209F inactive Saccharolobus solfataricus
V209G the mutant shows 60% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
V209K inactive Saccharolobus solfataricus
V209L the mutant shows 10% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
V209T the mutant shows 80% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
V209W inactive Saccharolobus solfataricus
W361A the mutant shows 241% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
W361F the mutant exhibits 4.2fold higher activity, 3.7fold higher catalytic efficiency, and 3.1fold lower binding energy for ginsenoside Rd than the wild type enzyme. The mutant completely converts ginsenoside Rb1 to compound K and shows also 7.4fold higher activity for hesperidin than the wild type enzyme Saccharolobus solfataricus
W361G the mutant shows about 290% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
W361L the mutant shows about 40% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
W361T the mutant shows about 350% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
W361Y the mutant shows about 230% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.52
-
 ginsenoside Rd wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
0.57
-
 ginsenoside Rb1 wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
1.59
-
 ginsenoside Rd mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
1.93
-
 ginsenoside Rb1 mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus P22498
-
-
Saccharolobus solfataricus DSM 1617 P22498
-
-

Purification (Commentary)

Purification (Comment) Organism
His-Trap column chromatography, and Superdex 200 gel filtration Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0019
-
 wild type enzyme, with ginsenoside C-Mc1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.0072
-
 mutant enzyme W361F, with ginsenoside C-Mc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.012
-
 mutant enzyme W361F, with ginsenoside C-Mc1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.0154
-
 wild type enzyme, with ginsenoside C-Mc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.0201
-
 wild type enzyme, with ginsenoside Rc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.0233
-
 mutant enzyme W361F, with ginsenoside Rc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.0355
-
 mutant enzyme W361F, with naringin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.0391
-
 wild type enzyme, with hesperidin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.233
-
 wild type enzyme, with naringin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.288
-
 mutant enzyme W361F, with hesperidin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
0.338
-
 wild type enzyme, with ginsenoside Rd as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
1.347
-
 mutant enzyme W361F, with ginsenoside C-Y as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
1.415
-
 mutant enzyme W361F, with ginsenoside Rd as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
1.643
-
 wild type enzyme, with ginsenoside Rb2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
2.031
-
 mutant enzyme W361F, with ginsenoside C-O as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.213
-
 mutant enzyme W361F, with ginsenoside Rb1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
4.388
-
 wild type enzyme, with ginsenoside C-Y as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
5.082
-
 wild type enzyme, with ginsenoside Rb1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
6.354
-
 wild type enzyme, with ginsenoside C-O as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
9.077
-
 mutant enzyme W361F, with ginsenoside Rb2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
13.13
-
 wild type enzyme, with ginsenoside F2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
13.227
-
 mutant enzyme W361F, with ginsenoside F2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-D-glucoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + D-glucose
-
 ?
4-nitrophenyl beta-D-glucoside + H2O
-
 Saccharolobus solfataricus DSM 1617 4-nitrophenol + D-glucose
-
 ?
ginsenoside C-Mc + H2O
-
 Saccharolobus solfataricus ?
-
 ?
ginsenoside C-Mc + H2O
-
 Saccharolobus solfataricus DSM 1617 ?
-
 ?
ginsenoside C-Mc1 + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinofuranose
-
 ?
ginsenoside C-Mc1 + H2O
-
 Saccharolobus solfataricus DSM 1617 ginsenoside C-K + D-glucose + L-arabinofuranose
-
 ?
ginsenoside C-O + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinopyranose
-
 ?
ginsenoside C-Y + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + L-arabinopyranose
-
 ?
ginsenoside F2 + H2O highest activity Saccharolobus solfataricus ginsenoside C-K + D-glucose
-
 ?
ginsenoside Rb1 + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose
-
 ?
ginsenoside Rb2 + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinopyranose
-
 ?
ginsenoside Rc + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinofuranose
-
 ?
ginsenoside Rd + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + ?
-
 ?
ginsenoside Rd + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose
-
 ?
hesperidin + H2O
-
 Saccharolobus solfataricus ?
-
 ?
hesperidin + H2O
-
 Saccharolobus solfataricus DSM 1617 ?
-
 ?
naringin + H2O
-
 Saccharolobus solfataricus ?
-
 ?
naringin + H2O
-
 Saccharolobus solfataricus DSM 1617 ?
-
 ?

Synonyms

Synonyms Comment Organism
beta-glycosidase
-
 Saccharolobus solfataricus
Bgl
-
 Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
92
-
 ginsenoside Rd wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
1049
-
 ginsenoside Rd mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
1343
-
 ginsenoside Rb1 mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
2257
-
 ginsenoside Rb1 wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
178
-
 ginsenoside Rd wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
660
-
 ginsenoside Rd mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
697
-
 ginsenoside Rb1 mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
3967
-
 ginsenoside Rb1 wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus