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Literature summary extracted from

  • Shin, K.C.; Hong, S.H.; Seo, M.J.; Oh, D.K.
    An amino acid at position 512 in beta-glucosidase from Clavibacter michiganensis determines the regioselectivity for hydrolyzing gypenoside XVII (2015), Appl. Microbiol. Biotechnol., 99, 7987-7996 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.191 recombinant expression of wild-type and mutant enzymes in Escherichia coli strain ER2566 Clavibacter michiganensis
3.2.1.193 recombinant expression of wild-type and mutant enzymes in Escherichia coli strain ER2566 Clavibacter michiganensis
3.2.1.194 recombinant expression of wild-type and mutant enzymes in Escherichia coli strain ER2566 Clavibacter michiganensis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.191 E332A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows decreased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 E83A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows increased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 H238A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows decreased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 L509A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows slightly decreased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 additional information the hydrolytic pathways of enzyme mutants E83A, R131A, R199A, H238A, E332A, and L509A convert GypXVII to GypLXXV, showing the same regioselectivity as the wild-type enzyme. But mutant W512A converts GypXVII to GypLXXV and F2 (cf. EC 3.2.1.193), indicating that the variant simultaneously hydrolyzes the inner glucose linked to the C-3 position and the outer glucose linked to the C-20 position in GypXVII. Mutant W512F variant enzyme containing an aromatic side chain converts GypXVII as a substrate to GypLXXV, while mutant W512R containing a positively charged side chain converts GypXVII to F2 Clavibacter michiganensis
3.2.1.191 R131A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows decreased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 R199A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows increased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 W331A site-directed mutagenesis, mutation of a potential catalytic residue, inactive mutant Clavibacter michiganensis
3.2.1.191 W512A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows slightly decreased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme. Mutant W512A converts GypXVII to GypLXXV and F2 (cf. EC 3.2.1.193) Clavibacter michiganensis
3.2.1.191 W512D site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows strongly decreased activity with ginsenoside Rb1 and no activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 W512E site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows decreased activity with ginsenoside Rb1 and no activity with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 W512F site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows slightly decreased activity with ginsenoside Rb1 and increased activity with gypenoside XVII compared to the wild-type enzyme. Mutant W512F variant enzyme containing an aromatic side chain converts GypXVII as a substrate to GypLXXV Clavibacter michiganensis
3.2.1.191 W512K site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows strongly decreased activity with ginsenoside Rb1 and with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.191 W512R site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows strongly decreased activity with ginsenoside Rb1 and with gypenoside XVII compared to the wild-type enzyme. mutant W512R containing a positively charged side chain converts GypXVII to F2, cf. EC 3.2.1.193 Clavibacter michiganensis
3.2.1.191 Y288A site-directed mutagenesis, mutation of a potential catalytic residue, inactive mutant Clavibacter michiganensis
3.2.1.193 additional information the hydrolytic pathways of enzyme mutants E83A, R131A, R199A, H238A, E332A, and L509A convert GypXVII to GypLXXV, showing the same regioselectivity as the wild-type enzyme. But mutant W512A converts GypXVII to GypLXXV and F2 (cf. EC 3.2.1.193), indicating that the variant simultaneously hydrolyzes the inner glucose linked to the C-3 position and the outer glucose linked to the C-20 position in GypXVII. Mutant W512F variant enzyme containing an aromatic side chain converts GypXVII as a substrate to GypLXXV, while mutant W512R containing a positively charged side chain converts GypXVII to F2 Clavibacter michiganensis
3.2.1.193 W512A site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows slightly decreased activity with ginsenoside Rb1 and decreased activity with gypenoside XVII compared to the wild-type enzyme. Mutant W512A converts GypXVII to GypLXXV and F2 (cf.. EC 3.2.1.193) Clavibacter michiganensis
3.2.1.193 W512K site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows strongly decreased activity with ginsenoside Rb1 and with gypenoside XVII compared to the wild-type enzyme Clavibacter michiganensis
3.2.1.193 W512R site-directed mutagenesis, mutation of a potential catalytic residue, the mutant shows strongly decreased activity with ginsenoside Rb1 and with gypenoside XVII compared to the wild-type enzyme. mutant W512R containing a positively charged side chain converts GypXVII to F2, cf. EC 3.2.1.193 Clavibacter michiganensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.191 0.18
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant wild-type enzyme Clavibacter michiganensis
3.2.1.191 0.2
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512A Clavibacter michiganensis
3.2.1.191 0.21
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512R Clavibacter michiganensis
3.2.1.191 0.32
-
 gypenoside XVII pH 7.5, 35°C, recombinant mutant W512R Clavibacter michiganensis
3.2.1.191 0.43
-
 gypenoside XVII pH 7.5, 35°C, recombinant mutant W512A Clavibacter michiganensis
3.2.1.191 0.46
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512E Clavibacter michiganensis
3.2.1.191 0.62
-
 gypenoside XVII pH 7.5, 35°C, recombinant wild-type enzyme Clavibacter michiganensis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.191 ginsenoside C-Mc1 + H2O Clavibacter michiganensis
-
 ginsenoside C-Mc + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside C-O + H2O Clavibacter michiganensis
-
 ginsenoside C-Y + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside F2 + H2O Clavibacter michiganensis
-
 ginsenoside C-K + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rb1 + H2O Clavibacter michiganensis
-
 gypenoside XVII + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rb1 + H2O Clavibacter michiganensis
-
 gypenoside XVII + ?
-
 ?
3.2.1.191 ginsenoside Rb2 + H2O Clavibacter michiganensis
-
 ginsenoside C-O + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rc + H2O Clavibacter michiganensis
-
 ginsenoside C-Mc1 + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rd + H2O Clavibacter michiganensis
-
 ginsenoside F2 + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rg3 + H2O Clavibacter michiganensis
-
 ginsenoside Rh2 + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rh2 + H2O Clavibacter michiganensis
-
 protopanaxadiol aglycone + D-glucopyranose
-
 ?
3.2.1.191 gypenoside LXXV + H2O Clavibacter michiganensis
-
 ginsenoside C-K + D-glucopyranose
-
 ?
3.2.1.191 gypenoside XVII + H2O Clavibacter michiganensis
-
 gypenoside LXXV + D-glucose
-
 ?
3.2.1.191 additional information Clavibacter michiganensis the specific activity of beta-glucosidase from Clavibacter michiganensis for the PPD-type ginsenosides as substrates followed the order Rb2 > Rd > Rc > Rg3 > Rb1 > compound O (C-O) > compound Mc1 (C-Mc1) > GypXVII > F2 > Rh2 > gypenside LXXV (GypLXXV) and the products of which are C-O, F2, C-Mc1, Rh2, GypXVII, compound Y (C-Y), compound Mc (C-Mc), GypLXXV, compound K (C-K), APPD, and C-K, respectively. The specific activity of beta-glucosidase from Clavibacter michiganensis for the PPT-type ginsenosides as substrates follows the order Rf > Rh1 > Rg1 and the products of which are Rh1, APPT, and F1, respectively ?
-
 ?
3.2.1.194 ginsenoside Rf + H2O Clavibacter michiganensis
-
 ginsenoside Rh1 + D-glucose
-
 ?
3.2.1.194 ginsenoside Rf + H2O Clavibacter michiganensis DSM 46364
-
 ginsenoside Rh1 + D-glucose
-
 ?
3.2.1.194 ginsenoside Rg1 + H2O Clavibacter michiganensis
-
 ginsenoside F1 + D-glucose
-
 ?
3.2.1.194 ginsenoside Rh1 + H2O Clavibacter michiganensis
-
 protopanaxatriol aglycone + D-glucose
-
 ?
3.2.1.194 additional information Clavibacter michiganensis the specific activity of beta-glucosidase from Clavibacter michiganensis for the PPD-type ginsenosides as substrates followed the order Rb2 > Rd > Rc > Rg3 > Rb1 > compound O (C-O) > compound Mc1 (C-Mc1) > GypXVII > F2 > Rh2 > gypenside LXXV (GypLXXV) and the products of which are C-O, F2, C-Mc1, Rh2, GypXVII, compound Y (C-Y), compound Mc (C-Mc), GypLXXV, compound K (C-K), APPD, and C-K, respectively. The specific activity of beta-glucosidase from Clavibacter michiganensis for the PPT-type ginsenosides as substrates followed the order Rf > Rh1 > Rg1 and the products of which are Rh1, APPT, and F1, respectively ?
-
 ?
3.2.1.194 additional information Clavibacter michiganensis DSM 46364 the specific activity of beta-glucosidase from Clavibacter michiganensis for the PPD-type ginsenosides as substrates followed the order Rb2 > Rd > Rc > Rg3 > Rb1 > compound O (C-O) > compound Mc1 (C-Mc1) > GypXVII > F2 > Rh2 > gypenside LXXV (GypLXXV) and the products of which are C-O, F2, C-Mc1, Rh2, GypXVII, compound Y (C-Y), compound Mc (C-Mc), GypLXXV, compound K (C-K), APPD, and C-K, respectively. The specific activity of beta-glucosidase from Clavibacter michiganensis for the PPT-type ginsenosides as substrates followed the order Rf > Rh1 > Rg1 and the products of which are Rh1, APPT, and F1, respectively ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.191 Clavibacter michiganensis
-
-
-
3.2.1.191 Clavibacter michiganensis DSM 46364
-
-
-
3.2.1.193 Clavibacter michiganensis
-
-
-
3.2.1.193 Clavibacter michiganensis DSM 46364
-
-
-
3.2.1.194 Clavibacter michiganensis
-
-
-
3.2.1.194 Clavibacter michiganensis DSM 46364
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.191 0.0136
-
 recombinant mutant W512K, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0196
-
 recombinant mutant W512A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0204
-
 recombinant mutant H238A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0273
-
 recombinant mutant R131A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0326
-
 recombinant mutant L509A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0333
-
 recombinant mutant W512R, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0335
-
 recombinant mutant R199A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0348
-
 recombinant mutant E332A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0354
-
 recombinant mutant E83A, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0421
-
 recombinant wild-type enzyme, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 0.0647
-
 recombinant mutant W512F, pH 7.5, 35°C, substrate gypenoside XVII Clavibacter michiganensis
3.2.1.191 1.173
-
 recombinant mutant W512K, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 1.248
-
 recombinant mutant W512D, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 1.474
-
 recombinant mutant W512R, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 1.665
-
 recombinant mutant H238A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 1.896
-
 recombinant mutant R131A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 1.998
-
 recombinant mutant E332A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2
-
 recombinant mutant W512E, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2.109
-
 recombinant mutant L509A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2.119
-
 recombinant mutant W512F, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2.135
-
 recombinant mutant W512A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2.189
-
 recombinant wild-type enzyme, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2.22
-
 recombinant mutant E83A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis
3.2.1.191 2.301
-
 recombinant mutant R199A, pH 7.5, 35°C, substrate ginsenoside Rb1 Clavibacter michiganensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.191 ginsenoside C-Mc1 + H2O
-
 Clavibacter michiganensis ginsenoside C-Mc + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside C-O + H2O
-
 Clavibacter michiganensis ginsenoside C-Y + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside F2 + H2O
-
 Clavibacter michiganensis ginsenoside C-K + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rb1 + H2O
-
 Clavibacter michiganensis gypenoside XVII + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rb1 + H2O
-
 Clavibacter michiganensis gypenoside XVII + ?
-
 ?
3.2.1.191 ginsenoside Rb2 + H2O
-
 Clavibacter michiganensis ginsenoside C-O + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rc + H2O
-
 Clavibacter michiganensis ginsenoside C-Mc1 + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rd + H2O
-
 Clavibacter michiganensis ginsenoside F2 + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rg3 + H2O
-
 Clavibacter michiganensis ginsenoside Rh2 + D-glucopyranose
-
 ?
3.2.1.191 ginsenoside Rh2 + H2O
-
 Clavibacter michiganensis protopanaxadiol aglycone + D-glucopyranose
-
 ?
3.2.1.191 gypenoside LXXV + H2O
-
 Clavibacter michiganensis ginsenoside C-K + D-glucopyranose
-
 ?
3.2.1.191 gypenoside LXXV + H2O GypLXXV, low activity Clavibacter michiganensis ginsenoside C-K + D-glucopyranose
-
 ?
3.2.1.191 gypenoside XVII + H2O
-
 Clavibacter michiganensis gypenoside LXXV + D-glucose
-
 ?
3.2.1.191 gypenoside XVII + H2O GypXVI, enzyme mutants W512A and W512K hydrolyze the inner glucose linked to the C-3 position and the outer glucose linked to the C-20 position of GypXVII to produce GypLXXV and F2. W512R hydrolyzes only the outer glucose linked to the C-20 position of GypXVII to produce F2. Mutants W512E and W512D exhibit no activity for GypXVII Clavibacter michiganensis gypenoside LXXV + D-glucose
-
 ?
3.2.1.191 additional information the specific activity of beta-glucosidase from Clavibacter michiganensis for the PPD-type ginsenosides as substrates followed the order Rb2 > Rd > Rc > Rg3 > Rb1 > compound O (C-O) > compound Mc1 (C-Mc1) > GypXVII > F2 > Rh2 > gypenside LXXV (GypLXXV) and the products of which are C-O, F2, C-Mc1, Rh2, GypXVII, compound Y (C-Y), compound Mc (C-Mc), GypLXXV, compound K (C-K), APPD, and C-K, respectively. The specific activity of beta-glucosidase from Clavibacter michiganensis for the PPT-type ginsenosides as substrates follows the order Rf > Rh1 > Rg1 and the products of which are Rh1, APPT, and F1, respectively Clavibacter michiganensis ?
-
 ?
3.2.1.191 additional information recombinant beta-glucosidase from Clavibacter michiganensis specifically hydrolyzes the outer and inner glucose linked to the C-3 position in protopanaxadiol (PPD)-type ginsenosides (EC 3.2.1.191) and the C-6 position in protopanaxatriol (PPT)-type ginsenosides (EC 3.2.1.194) except for the hydrolysis of gypenoside LXXV (GypLXXV). The enzyme converts gypenoside XVII (GypXVII) to GypLXXV by hydrolyzing the inner glucose linked to the C-3 position Clavibacter michiganensis ?
-
 ?
3.2.1.193 gypenoside XVII + H2O GypXVI, enzyme mutants W512A and W512K hydrolyze the inner glucose linked to the C-3 position and the outer glucose linked to the C-20 position of GypXVII to produce GypLXXV (cf. EC 3.2.1.191) and F2. Mutant W512R hydrolyzes only the outer glucose linked to the C-20 position of GypXVII to produce F2. The wild-type enzyme hydrolyzes GypXVI only to gypenoside LXXV, GypLXXV Clavibacter michiganensis F2 + D-glucose
-
 ?
3.2.1.193 gypenoside XVII + H2O GypXVI, enzyme mutants W512A and W512K hydrolyze the inner glucose linked to the C-3 position and the outer glucose linked to the C-20 position of GypXVII to produce GypLXXV (cf. EC 3.2.1.191) and F2. Mutant W512R hydrolyzes only the outer glucose linked to the C-20 position of GypXVII to produce F2. The wild-type enzyme hydrolyzes GypXVI only to gypenoside LXXV, GypLXXV Clavibacter michiganensis DSM 46364 F2 + D-glucose
-
 ?
3.2.1.193 additional information recombinant wild-type beta-glucosidase from Clavibacter michiganensis specifically hydrolyzes the outer and inner glucose linked to the C-3 position in protopanaxadiol (PPD)-type ginsenosides (EC 3.2.1.191) and the C-6 position in protopanaxatriol (PPT)-type ginsenosides (EC 3.2.1.194) except for the hydrolysis of gypenoside LXXV (GypLXXV). The enzyme converts gypenoside XVII (GypXVII) to GypLXXV by hydrolyzing the inner glucose linked to the C-3 position Clavibacter michiganensis ?
-
 ?
3.2.1.193 additional information recombinant wild-type beta-glucosidase from Clavibacter michiganensis specifically hydrolyzes the outer and inner glucose linked to the C-3 position in protopanaxadiol (PPD)-type ginsenosides (EC 3.2.1.191) and the C-6 position in protopanaxatriol (PPT)-type ginsenosides (EC 3.2.1.194) except for the hydrolysis of gypenoside LXXV (GypLXXV). The enzyme converts gypenoside XVII (GypXVII) to GypLXXV by hydrolyzing the inner glucose linked to the C-3 position Clavibacter michiganensis DSM 46364 ?
-
 ?
3.2.1.194 ginsenoside Rf + H2O
-
 Clavibacter michiganensis ginsenoside Rh1 + D-glucose
-
 ?
3.2.1.194 ginsenoside Rf + H2O
-
 Clavibacter michiganensis DSM 46364 ginsenoside Rh1 + D-glucose
-
 ?
3.2.1.194 ginsenoside Rg1 + H2O
-
 Clavibacter michiganensis ginsenoside F1 + D-glucose
-
 ?
3.2.1.194 ginsenoside Rh1 + H2O
-
 Clavibacter michiganensis protopanaxatriol aglycone + D-glucose
-
 ?
3.2.1.194 additional information the specific activity of beta-glucosidase from Clavibacter michiganensis for the PPD-type ginsenosides as substrates followed the order Rb2 > Rd > Rc > Rg3 > Rb1 > compound O (C-O) > compound Mc1 (C-Mc1) > GypXVII > F2 > Rh2 > gypenside LXXV (GypLXXV) and the products of which are C-O, F2, C-Mc1, Rh2, GypXVII, compound Y (C-Y), compound Mc (C-Mc), GypLXXV, compound K (C-K), APPD, and C-K, respectively. The specific activity of beta-glucosidase from Clavibacter michiganensis for the PPT-type ginsenosides as substrates followed the order Rf > Rh1 > Rg1 and the products of which are Rh1, APPT, and F1, respectively Clavibacter michiganensis ?
-
 ?
3.2.1.194 additional information recombinant beta-glucosidase from Clavibacter michiganensis specifically hydrolyzes the outer and inner glucose linked to the C-3 position in protopanaxadiol (PPD)-type ginsenosides (EC 3.2.1.191) and the C-6 position in protopanaxatriol (PPT)-type ginsenosides (EC 3.2.1.194) except for the hydrolysis of gypenoside LXXV (GypLXXV). The enzyme converts gypenoside XVII (GypXVII) to GypLXXV by hydrolyzing the inner glucose linked to the C-3 position Clavibacter michiganensis ?
-
 ?
3.2.1.194 additional information the specific activity of beta-glucosidase from Clavibacter michiganensis for the PPD-type ginsenosides as substrates followed the order Rb2 > Rd > Rc > Rg3 > Rb1 > compound O (C-O) > compound Mc1 (C-Mc1) > GypXVII > F2 > Rh2 > gypenside LXXV (GypLXXV) and the products of which are C-O, F2, C-Mc1, Rh2, GypXVII, compound Y (C-Y), compound Mc (C-Mc), GypLXXV, compound K (C-K), APPD, and C-K, respectively. The specific activity of beta-glucosidase from Clavibacter michiganensis for the PPT-type ginsenosides as substrates followed the order Rf > Rh1 > Rg1 and the products of which are Rh1, APPT, and F1, respectively Clavibacter michiganensis DSM 46364 ?
-
 ?
3.2.1.194 additional information recombinant beta-glucosidase from Clavibacter michiganensis specifically hydrolyzes the outer and inner glucose linked to the C-3 position in protopanaxadiol (PPD)-type ginsenosides (EC 3.2.1.191) and the C-6 position in protopanaxatriol (PPT)-type ginsenosides (EC 3.2.1.194) except for the hydrolysis of gypenoside LXXV (GypLXXV). The enzyme converts gypenoside XVII (GypXVII) to GypLXXV by hydrolyzing the inner glucose linked to the C-3 position Clavibacter michiganensis DSM 46364 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.191 More cf. EC 3.2.1.195, EC 3.2.1.193, and EC 3.2.1.194 Clavibacter michiganensis
3.2.1.193 More cf. EC 3.2.1.191 and EC 3.2.1.194 Clavibacter michiganensis
3.2.1.194 More cf. EC 3.2.1.191 Clavibacter michiganensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.191 35
-
-
 Clavibacter michiganensis
3.2.1.193 35
-
-
 Clavibacter michiganensis
3.2.1.194 35
-
-
 Clavibacter michiganensis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.191 9
-
 gypenoside XVII pH 7.5, 35°C, recombinant mutant W512A Clavibacter michiganensis
3.2.1.191 9
-
 gypenoside XVII pH 7.5, 35°C, recombinant mutant W512R Clavibacter michiganensis
3.2.1.191 22
-
 gypenoside XVII pH 7.5, 35°C, recombinant wild-type enzyme Clavibacter michiganensis
3.2.1.191 343
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512R Clavibacter michiganensis
3.2.1.191 795
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512A Clavibacter michiganensis
3.2.1.191 1025
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant wild-type enzyme Clavibacter michiganensis
3.2.1.191 1151
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512E Clavibacter michiganensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.191 7.5
-
-
 Clavibacter michiganensis
3.2.1.193 7.5
-
-
 Clavibacter michiganensis
3.2.1.194 7.5
-
-
 Clavibacter michiganensis

General Information

EC Number General Information Comment Organism
3.2.1.191 additional information the amino acid at position 512 in beta-glucosidase from Clavibacter michiganensis determines the regioselectivity for hydrolyzing gypenoside XVII, obtained from the GypXVII-docked homology models of beta-glucosidase. Homology modeling of Clavibacter michiganensis beta-glucosidase based on the crystal structure of beta-D-glucan exohydrolase from Hordeum vulgare (PDB ID 1EX1) as a template Clavibacter michiganensis
3.2.1.193 additional information the amino acid at position 512 in beta-glucosidase from Clavibacter michiganensis determines the regioselectivity for hydrolyzing gypenoside XVII, obtained from the GypXVII-docked homology models of beta-glucosidase. Homology modeling of Clavibacter michiganensis beta-glucosidase based on the crystal structure of beta-D-glucan exohydrolase from Hordeum vulgare (PDB ID 1EX1) as a template Clavibacter michiganensis
3.2.1.194 additional information the amino acid at position 512 in beta-glucosidase from Clavibacter michiganensis determines the regioselectivity for hydrolyzing gypenoside XVII, obtained from the GypXVII-docked homology models of beta-glucosidase. Homology modeling of Clavibacter michiganensis beta-glucosidase based on the crystal structure of beta-D-glucan exohydrolase from Hordeum vulgare (PDB ID 1EX1) as a template Clavibacter michiganensis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.191 20.9
-
 gypenoside XVII pH 7.5, 35°C, recombinant mutant W512A Clavibacter michiganensis
3.2.1.191 28.125
-
 gypenoside XVII pH 7.5, 35°C, recombinant mutant W512R Clavibacter michiganensis
3.2.1.191 35.5
-
 gypenoside XVII pH 7.5, 35°C, recombinant wild-type enzyme Clavibacter michiganensis
3.2.1.191 1633
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512R Clavibacter michiganensis
3.2.1.191 2502
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512E Clavibacter michiganensis
3.2.1.191 3975
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant mutant W512A Clavibacter michiganensis
3.2.1.191 5694
-
 ginsenoside Rb1 pH 7.5, 35°C, recombinant wild-type enzyme Clavibacter michiganensis