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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.B26 expressed in Escherichia coli strain ER2566 Saccharolobus solfataricus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.B26 F222A the mutant shows 105% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 F359A the mutant shows 84% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 L213A the mutant shows 110% activity with ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 S220A the mutant shows 92% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 V209A the mutant shows 170% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 V209D inactive Saccharolobus solfataricus
3.2.1.B26 V209F inactive Saccharolobus solfataricus
3.2.1.B26 V209G the mutant shows 60% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 V209K inactive Saccharolobus solfataricus
3.2.1.B26 V209L the mutant shows 10% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 V209T the mutant shows 80% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 V209W inactive Saccharolobus solfataricus
3.2.1.B26 W361A the mutant shows 241% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 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
3.2.1.B26 W361G the mutant shows about 290% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 W361L the mutant shows about 40% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 W361T the mutant shows about 350% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus
3.2.1.B26 W361Y the mutant shows about 230% activity towards ginsenoside Rd compared to the wild type enzyme Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.B26 0.52
-
 ginsenoside Rd wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.57
-
 ginsenoside Rb1 wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1.59
-
 ginsenoside Rd mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1.93
-
 ginsenoside Rb1 mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.B26 Saccharolobus solfataricus P22498
-
-
3.2.1.B26 Saccharolobus solfataricus DSM 1617 P22498
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.B26 His-Trap column chromatography, and Superdex 200 gel filtration Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.B26 0.0019
-
 wild type enzyme, with ginsenoside C-Mc1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.0072
-
 mutant enzyme W361F, with ginsenoside C-Mc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.012
-
 mutant enzyme W361F, with ginsenoside C-Mc1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.0154
-
 wild type enzyme, with ginsenoside C-Mc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.0201
-
 wild type enzyme, with ginsenoside Rc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.0233
-
 mutant enzyme W361F, with ginsenoside Rc as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.0355
-
 mutant enzyme W361F, with naringin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.0391
-
 wild type enzyme, with hesperidin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.233
-
 wild type enzyme, with naringin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.288
-
 mutant enzyme W361F, with hesperidin as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 0.338
-
 wild type enzyme, with ginsenoside Rd as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1.347
-
 mutant enzyme W361F, with ginsenoside C-Y as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1.415
-
 mutant enzyme W361F, with ginsenoside Rd as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1.643
-
 wild type enzyme, with ginsenoside Rb2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 2.031
-
 mutant enzyme W361F, with ginsenoside C-O as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 3.213
-
 mutant enzyme W361F, with ginsenoside Rb1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 4.388
-
 wild type enzyme, with ginsenoside C-Y as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 5.082
-
 wild type enzyme, with ginsenoside Rb1 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 6.354
-
 wild type enzyme, with ginsenoside C-O as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 9.077
-
 mutant enzyme W361F, with ginsenoside Rb2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 13.13
-
 wild type enzyme, with ginsenoside F2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 13.227
-
 mutant enzyme W361F, with ginsenoside F2 as substrate, at pH 5.5 and 95°C Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.B26 4-nitrophenyl beta-D-glucoside + H2O
-
 Saccharolobus solfataricus 4-nitrophenol + D-glucose
-
 ?
3.2.1.B26 4-nitrophenyl beta-D-glucoside + H2O
-
 Saccharolobus solfataricus DSM 1617 4-nitrophenol + D-glucose
-
 ?
3.2.1.B26 ginsenoside C-Mc + H2O
-
 Saccharolobus solfataricus ?
-
 ?
3.2.1.B26 ginsenoside C-Mc + H2O
-
 Saccharolobus solfataricus DSM 1617 ?
-
 ?
3.2.1.B26 ginsenoside C-Mc1 + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinofuranose
-
 ?
3.2.1.B26 ginsenoside C-Mc1 + H2O
-
 Saccharolobus solfataricus DSM 1617 ginsenoside C-K + D-glucose + L-arabinofuranose
-
 ?
3.2.1.B26 ginsenoside C-O + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinopyranose
-
 ?
3.2.1.B26 ginsenoside C-Y + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + L-arabinopyranose
-
 ?
3.2.1.B26 ginsenoside F2 + H2O highest activity Saccharolobus solfataricus ginsenoside C-K + D-glucose
-
 ?
3.2.1.B26 ginsenoside Rb1 + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose
-
 ?
3.2.1.B26 ginsenoside Rb2 + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinopyranose
-
 ?
3.2.1.B26 ginsenoside Rc + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose + L-arabinofuranose
-
 ?
3.2.1.B26 ginsenoside Rd + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + ?
-
 ?
3.2.1.B26 ginsenoside Rd + H2O
-
 Saccharolobus solfataricus ginsenoside C-K + D-glucose
-
 ?
3.2.1.B26 hesperidin + H2O
-
 Saccharolobus solfataricus ?
-
 ?
3.2.1.B26 hesperidin + H2O
-
 Saccharolobus solfataricus DSM 1617 ?
-
 ?
3.2.1.B26 naringin + H2O
-
 Saccharolobus solfataricus ?
-
 ?
3.2.1.B26 naringin + H2O
-
 Saccharolobus solfataricus DSM 1617 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.B26 beta-glycosidase
-
 Saccharolobus solfataricus
3.2.1.B26 Bgl
-
 Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.B26 92
-
 ginsenoside Rd wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1049
-
 ginsenoside Rd mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 1343
-
 ginsenoside Rb1 mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 2257
-
 ginsenoside Rb1 wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus

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.B26 178
-
 ginsenoside Rd wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 660
-
 ginsenoside Rd mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 697
-
 ginsenoside Rb1 mutant enzyme W361F, at pH 5.5 and 95°C Saccharolobus solfataricus
3.2.1.B26 3967
-
 ginsenoside Rb1 wild type enzyme, at pH 5.5 and 95°C Saccharolobus solfataricus