3.2.1.B26: Sulfolobus solfataricus beta-glycosidase
This is an abbreviated version!
For detailed information about Sulfolobus solfataricus beta-glycosidase, go to the full flat file.
Word Map on EC 3.2.1.B26
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3.2.1.B26
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transglycosylation
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synthesis
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beta-d-galactoside
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beta-glucosides
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amygdalin
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glycone
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3.2.1.21
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transgalactosylation
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prunasin
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beta-d-fucoside
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food industry
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analysis
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nutrition
- 3.2.1.B26
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transglycosylation
- synthesis
- beta-d-galactoside
- beta-glucosides
- amygdalin
-
glycone
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3.2.1.21
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transgalactosylation
- prunasin
- beta-d-fucoside
- food industry
- analysis
- nutrition
Reaction
Wide substrate specificity, active on aryl-beta-D-galactose, -alpha-L-fucose, -beta-D-glucose and -beta-D-xylose and on di- and oligosaccharides. D-Glucose dimers are hydrolysed in the order of decreasing efficiency: beta-(1,3), beta-(1,4), beta-(1,6). Exo-acting enzyme with a preference for cellotetraose. =
Synonyms
beta-D-glycosidase, beta-glucosidase, beta-Gly, beta-glycosidase, Bgl, bgly, EcSbgly, GH1 beta-glycosidase, LACS, Sbeta-gly, Sbetagly, Ss-beta-Gly, Ssbeta-Glc1, Ssbeta-Gly, SsbetaG, SsbetaGlc1, SsbetaGly, SsGH1, sso1353, SSO3019
ECTree
3.2.1.B26 Sulfolobus solfataricus beta-glycosidaseAdvanced search results
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results (Engineering
Engineering on EC 3.2.1.B26 - Sulfolobus solfataricus beta-glycosidase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
D462G
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inactive mutant. Addition of 1 mM NaN3 rescues enzymatic activity using 2-nitrophenyl-beta-D-glucopyranoside as substrate. Activity is 7fold lower compared to wild-type
delHis489
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mutation produces faster enzyme inactivation, kcat/KM for 2-nitrophenyl beta-D-galactopyranoside is 45% of the wild-type value
delVal484-His489
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clone DELTA6 lacks the last six amino acids (-Val-Lys-Pro-Leu-Arg-His-COOH) and has no additional mutations. Mutation produces faster enzyme inactivation, kcat/KM for 2-nitrophenyl beta-D-galactopyranoside is 37% of the wild-type value
E206Q
E386G
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mutation in nucleophile residue E387, mutation completely abolishes activity under statndard conditions. The addition of 2 M sodium formate as an external nucleophile leads to the recovery of 8.40% activity with accumulation of oligosaccharides. At pH 3.0 and low concentrations of sodium formate buffer, the hyperthermophilic glycosynthase shows kcat values similar to those of the wild-type and 17fold higher than those observed at the usual reactivation conditions in 2 M sodium formate at pH 6.5
E387A
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inactive mutant enzyme of the catalytic nucleophile Glu387 is restored by externally added nucleophiles (sodium azide and sodium formate)
E387G
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inactive mutant enzyme of the catalytic nucleophile Glu387 is restored by externally added nucleophiles (sodium azide and sodium formate)
E387Q
E432C
F222A
the mutant shows 105% activity towards ginsenoside Rd compared to the wild type enzyme
F359A
the mutant shows 84% activity towards ginsenoside Rd compared to the wild type enzyme
H342A
the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol)
H489A
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mutation produces faster enzyme inactivation, kcat/KM for 2-nitrophenyl beta-D-galactopyranoside is 85% of the wild-type value
K219R
the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol)
L213A
L213Q
the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol)
L213W
the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol)
M439C
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shows almost identical values to wild-type for 4-methylumbelliferyl beta-D-glucopyranoside, 4-methylumbelliferyl beta-D-galactopyranoside or 4-methylumbelliferyl beta-D-mannopyranoside substrates. kcat/KM for 4-methylumbelliferyl beta-D-fucopyranoside is 1.8-fold lower than wild-type value and kcat/KM for 4-methylumbelliferyl beta-D-xylopyranoside is 4.7fold higher than wild-type
N97C
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mutation causes some changes in the structural and dynamic properties as observed by circular dichroism in far- and near-UV light, as well as by frequency domain fluorometry, with a simultaneous loss of thermostability
R488A
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mutation produces faster enzyme inactivation, kcat/KM for 2-nitrophenyl beta-D-galactopyranoside is 90% of the wild-type value
S101C
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mutation causes some changes in the structural and dynamic properties as observed by circular dichroism in far- and near-UV light, as well as by frequency domain fluorometry, with a simultaneous loss of thermostability
S220A
the mutant shows 92% activity towards ginsenoside Rd compared to the wild type enzyme
V209A
the mutant shows 170% activity towards ginsenoside Rd compared to the wild type enzyme
V209G
the mutant shows 60% activity towards ginsenoside Rd compared to the wild type enzyme
V209L
the mutant shows 10% activity towards ginsenoside Rd compared to the wild type enzyme
V209T
the mutant shows 80% activity towards ginsenoside Rd compared to the wild type enzyme
W361A
the mutant shows 241% activity towards ginsenoside Rd compared to the wild type enzyme
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
W361G
the mutant shows about 290% activity towards ginsenoside Rd compared to the wild type enzyme
W361L
the mutant shows about 40% activity towards ginsenoside Rd compared to the wild type enzyme
W361T
the mutant shows about 350% activity towards ginsenoside Rd compared to the wild type enzyme
W361Y
the mutant shows about 230% activity towards ginsenoside Rd compared to the wild type enzyme
W433C
F222A
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the mutant shows 105% activity towards ginsenoside Rd compared to the wild type enzyme
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F359A
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the mutant shows 84% activity towards ginsenoside Rd compared to the wild type enzyme
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K219R
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the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol)
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L213A
L213W
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the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol)
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S220A
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the mutant shows 92% activity towards ginsenoside Rd compared to the wild type enzyme
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W361A
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the mutant shows 241% activity towards ginsenoside Rd compared to the wild type enzyme
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E206Q
E432C
W433C
additional information
E206Q
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mutant shows 10- and 60-fold reduced activities on aryl-galacto and aryl-glucosides, respectively, when compared with the wild type. Significant decrease in Km-value with 4-nitrophenyl beta-D-glucoside or 2-nitrophenyl beta-D-glucoside. The residual activity of the mutant loses the typical pH dependence shown by the wild type. These data suggest that Glu206 acts as the general acid/base catalyst in the hydrolysis reaction
E206Q
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mutant shows 10- and 60-fold reduced activities on aryl-galacto and aryl-glucosides, respectively, when compared with the wild type. Significant decrease in Km-value with 4-nitrophenyl beta-D-glucoside or 2-nitrophenyl betaD-glucoside. The residual activity of the mutant loses the typical pH dependence shown by the wild type. These data suggest that Glu206 acts as the general acid/base catalyst in the hydrolysis reaction
E432C
kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 200fold, kcat/KM for 4-methylumbelliferyl beta-D-galactoside is reduced 130fold, kcat/KM for 4-methylumbelliferyl beta-D-xyloside is reduced 1.5fold, kcat/KM for 4-methylumbelliferyl beta-D-fucoside is reduced 2900fold, kcat/KM for 4-methylumbelliferyl beta-D-mannoside is ibcreased 1.5fold
E432C
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kcat/KM is reduced for 4-methylumbelliferyl beta-D-glucopyranoside is reduced 200fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-galactopyranoside is reduced 130fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-mannopyranoside is reduced 16fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-xylopyranoside is reduced 14fold
E432C
the kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 200fold, and 4-methylumbelliferyl beta-D-galactoside is reduced 130fold. Although the activities with 4-methylumbelliferyl beta-D-mannoside and 4-methylumbelliferyl beta-D-xyloside are also reduced in the mutant, this is to a much smaller extent. The mutant enzyme shows remarkably broader specificity than wild-type enzyme
L213A
the mutant shows 110% activity with ginsenoside Rd compared to the wild type enzyme
L213A
the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol). The variant enzyme converts ginsenosides Rc and C-Mc into C-K with molar conversions of 97%, which are 1.5 and 2fold higher, respectively, than those of the wild type enzyme
W433C
kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 140fold, kcat/KM for 4-methylumbelliferyl beta-D-galactoside is reduced 230fold, kcat/KM for 4-methylumbelliferyl beta-D-xyloside is reduced 14fold, kcat/KM for 4-methylumbelliferyl beta-D-fucoside is reduced 138fold, kcat/KM for 4-methylumbelliferyl beta-D-mannoside is reduced 16fold
W433C
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kcat/KM is reduced for 4-methylumbelliferyl beta-D-glucopyranoside is reduced 140fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-galactopyranoside is reduced 230fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-mannopyranoside is reduced 10fold, kcat/KM is reduced for 4-methylumbelliferyl beta-D-xylopyranoside is reduced 12fold
W433C
the kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 140fold, and 4-methylumbelliferyl beta-D-galactoside is reduced 230fold. Although the activities with 4-methylumbelliferyl beta-D-mannoside and 4-methylumbelliferyl beta-D-xyloside are also reduced in the mutant, this is to a much smaller extent. The mutant enzyme shows remarkably broader specificity than wild-type enzyme
L213A
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the mutant with increased alpha-L-arabinofuranosidase activity converts ginsenoside Rc to ginsenoside C-K (i.e. 20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol). The variant enzyme converts ginsenosides Rc and C-Mc into C-K with molar conversions of 97%, which are 1.5 and 2fold higher, respectively, than those of the wild type enzyme
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L213A
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the mutant shows 110% activity with ginsenoside Rd compared to the wild type enzyme
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E206Q
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mutant shows 10- and 60-fold reduced activities on aryl-galacto and aryl-glucosides, respectively, when compared with the wild type. Significant decrease in Km-value with 4-nitrophenyl beta-D-glucoside or 2-nitrophenyl beta-D-glucoside. The residual activity of the mutant loses the typical pH dependence shown by the wild type. These data suggest that Glu206 acts as the general acid/base catalyst in the hydrolysis reaction
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E206Q
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mutant shows 10- and 60-fold reduced activities on aryl-galacto and aryl-glucosides, respectively, when compared with the wild type. Significant decrease in Km-value with 4-nitrophenyl beta-D-glucoside or 2-nitrophenyl betaD-glucoside. The residual activity of the mutant loses the typical pH dependence shown by the wild type. These data suggest that Glu206 acts as the general acid/base catalyst in the hydrolysis reaction
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E432C
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kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 200fold, kcat/KM for 4-methylumbelliferyl beta-D-galactoside is reduced 130fold, kcat/KM for 4-methylumbelliferyl beta-D-xyloside is reduced 1.5fold, kcat/KM for 4-methylumbelliferyl beta-D-fucoside is reduced 2900fold, kcat/KM for 4-methylumbelliferyl beta-D-mannoside is ibcreased 1.5fold
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E432C
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the kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 200fold, and 4-methylumbelliferyl beta-D-galactoside is reduced 130fold. Although the activities with 4-methylumbelliferyl beta-D-mannoside and 4-methylumbelliferyl beta-D-xyloside are also reduced in the mutant, this is to a much smaller extent. The mutant enzyme shows remarkably broader specificity than wild-type enzyme
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W433C
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kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 140fold, kcat/KM for 4-methylumbelliferyl beta-D-galactoside is reduced 230fold, kcat/KM for 4-methylumbelliferyl beta-D-xyloside is reduced 14fold, kcat/KM for 4-methylumbelliferyl beta-D-fucoside is reduced 138fold, kcat/KM for 4-methylumbelliferyl beta-D-mannoside is reduced 16fold
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W433C
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the kcat/KM for 4-methylumbelliferyl beta-D-glucoside is reduced 140fold, and 4-methylumbelliferyl beta-D-galactoside is reduced 230fold. Although the activities with 4-methylumbelliferyl beta-D-mannoside and 4-methylumbelliferyl beta-D-xyloside are also reduced in the mutant, this is to a much smaller extent. The mutant enzyme shows remarkably broader specificity than wild-type enzyme
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additional information
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isolation of mutants with reduced expression of lacS. The mechanism of control occurs at the level of transcription. The mutations result in undetectable levels of lacS mRNA. The mechanism of reduction in lacS mRNA levels is unknown. It may result from events that act at the level of mRNA synthesis or mRNA degradation. Glycosyl hydrolase production in Sulfolobuis solfataricus requires a trans-acting factor for wild-type levels of expression. The trans-acting factor must recognize elements inherent to all target genes
additional information
generation of functional glycoside hydrolase hybrids by shuffling of the genes coding for Pyrococcus furiosus CelB and Sulfolobus solfataricus LacS. Generating of thermostable hybrid beta-glycosidases and isolating high-performance variants
additional information
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isolation of mutants with reduced expression of lacS. The mechanism of control occurs at the level of transcription. The mutations result in undetectable levels of lacS mRNA. The mechanism of reduction in lacS mRNA levels is unknown. It may result from events that act at the level of mRNA synthesis or mRNA degradation. Glycosyl hydrolase production in Sulfolobuis solfataricus requires a trans-acting factor for wild-type levels of expression. The trans-acting factor must recognize elements inherent to all target genes
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additional information
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generation of functional glycoside hydrolase hybrids by shuffling of the genes coding for Pyrococcus furiosus CelB and Sulfolobus solfataricus LacS. Generating of thermostable hybrid beta-glycosidases and isolating high-performance variants
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