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Literature summary extracted from

  • Li, B.; Ji, Y.; Li, Y.; Ding, G.
    Characterization of a glycoside hydrolase family 78 alpha-L-rhamnosidase from Bacteroides the taiotaomicron VPI-5482 and identification of functional residues (2018), 3 Biotech, 8, 120 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
3.2.1.40 industry the enzyme is a promising alternative biocatalyst for industrial applications due to good pH stability, relatively high thermostability and tolerance to low concentration of alcohols Bacteroides thetaiotaomicron

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.40 overexpression in Escherichia coli BL21(DE3) Bacteroides thetaiotaomicron

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.40 D330A 2350fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 D335A 560fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 D335E 2770fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 D335N 1170fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 D342A 7670fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 E595A 6970fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 E595Q 19170fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 H620A 40fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 R334A 280fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 W339A 1230fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 W440A 530fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron
3.2.1.40 Y383A 740fold reduction in specific activity as compared to the wild-type enzyme Bacteroides thetaiotaomicron

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.40 acetone 1% (v/v), about 50% loss of activity, 5% (v/v), about 75% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 acetonitrile 1% (v/v), about 55% loss of activity, 5% (v/v), about 90% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 DMSO 1% (v/v), about 45% loss of activity, 5% (v/v), about 75% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 ethanol 1% (v/v), about 25% loss of activity, 5% (v/v), about 55% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 ethylene glycol 1% (v/v), about 15% loss of activity, 5% (v/v), about 30% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 Isopropanol 1% (v/v), about 25% loss of activity, 5% (v/v), about 55% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 methanol 1% (v/v), about 15% loss of activity, 5% (v/v), about 50% loss of activity Bacteroides thetaiotaomicron
3.2.1.40 N,N-Dimethylformamide 1% (v/v), about 70% loss of activity, 5% (v/v), about 90% loss of activity Bacteroides thetaiotaomicron

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.40 0.53
-
 4-nitrophenyl alpha-L-rhamnopyranoside 50°C, pH 6.5 Bacteroides thetaiotaomicron

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.40 86000
-
 SDS-PAGE Bacteroides thetaiotaomicron

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.40 Bacteroides thetaiotaomicron Q8A916
-
-
3.2.1.40 Bacteroides thetaiotaomicron VPI-5482 Q8A916
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.40
-
 Bacteroides thetaiotaomicron

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.40 4-nitrophenyl alpha-L-rhamnopyranoside + H2O
-
 Bacteroides thetaiotaomicron 4-nitrophenol + alpha-L-rhamnopyranose
-
 ?
3.2.1.40 4-nitrophenyl alpha-L-rhamnopyranoside + H2O
-
 Bacteroides thetaiotaomicron VPI-5482 4-nitrophenol + alpha-L-rhamnopyranose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.40 ? x * 86000, SDS-PAGE Bacteroides thetaiotaomicron

Synonyms

EC Number Synonyms Comment Organism
3.2.1.40 BtRha78A
-
 Bacteroides thetaiotaomicron

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.40 60
-
-
 Bacteroides thetaiotaomicron

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.40 35 65 35°C: about 50% of maximal activity, 65°C: about 70% of maximal activity Bacteroides thetaiotaomicron

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.40 50
-
 half-life: approximately 100 min Bacteroides thetaiotaomicron

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.40 69.8
-
 4-nitrophenyl alpha-L-rhamnopyranoside 50°C, pH 6.5 Bacteroides thetaiotaomicron

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.40 6.5
-
-
 Bacteroides thetaiotaomicron

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.40 5.5 8.6 pH 5.5: about 80% of maximal activity, high activity (more than 60%) at alkaline pH 7.5-8.6 Bacteroides thetaiotaomicron

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.40 5
-
 1 h, 40 min, enzyme retains 75% of its initial activity Bacteroides thetaiotaomicron
3.2.1.40 5.5 9 1 h, 40°C, more than 95% of the activity is retained Bacteroides thetaiotaomicron
3.2.1.40 8.6 9 the activity of BtRha78A is enhanced significantly after the incubation in alkaline environment. The enzyme retains 142% and 133% of initial activity after the incubation of 1 h at 40 °C in 50 mM glycine-NaOH buffer (pH 8.6 and pH 9.0), respectively Bacteroides thetaiotaomicron

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.40 130
-
 4-nitrophenyl alpha-L-rhamnopyranoside 50°C, pH 6.5 Bacteroides thetaiotaomicron