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Literature summary extracted from

  • Liu, W.; Lin, Y.; Jeng, W.; Chen, J.; Wang, A.; Shyur, L.
    Engineering of dual-functional hybrid glucanases (2012), Protein Eng. Des. Sel., 25, 771-780.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.39 gene TmLam, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Thermotoga maritima
3.2.1.73 expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Fibrobacter succinogenes

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.39 additional information engineering of dual-functional hybrid glucanases from a truncated and mutated 1,3-1,4-beta-D-glucanase gene TFsW203F from Fibrobacter succinogenes, and a 1,3-beta-D-glucanase gene TmLam from hyperthermophilic Thermotoga maritima used as target enzymes, by ligating substrate-binding domains (TmB1 and TmB2) and the catalytic domain (TmLamCD) of TmLam to the N- or C-terminus of TFsW203F to create four hybrid enzymes, TmB1-TFsW203F, TFsW203F-TmB2, TmB1-TFsW203F-TmB2 and TFsW203F-TmLamCD, creation of desirable hybrid enzymes with economic benefits for industrial applications. Improved thermal tolerance of the hybrid enzyme TFsW203FTmLamCD, fluorescence and circular dichroism spectrometric analyses, overview. Kinetic properties of mutant hybrid glucanases Thermotoga maritima
3.2.1.73 additional information engineering of dual-functional hybrid glucanases from a truncated and mutated 1,3-1,4-beta-D-glucanase gene TFsW203F from Fibrobacter succinogenes, and a 1,3-beta-D-glucanase gene TmLam from hyperthermophilic Thermotoga maritima used as target enzymes, by ligating substrate-binding domains (TmB1 and TmB2) and the catalytic domain (TmLamCD) of TmLam to the N- or C-terminus of TFsW203F to create four hybrid enzymes, TmB1-TFsW203F, TFsW203F-TmB2, TmB1-TFsW203F-TmB2 and TFsW203F-TmLamCD, creation of desirable hybrid enzymes with economic benefits for industrial applications. Improved thermal tolerance of the hybrid enzyme TFsW203FTmLamCD, fluorescence and circular dichroism spectrometric analyses, overview. Kinetic properties of parental TFsW203F and mutant hybrid glucanases Fibrobacter succinogenes
3.2.1.73 W203F site-directed mutagenesis, truncated and mutated 1,3–1,4-beta-D-glucanase, no activity with laminarin Fibrobacter succinogenes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.39 additional information
-
 additional information 8.3 mg/ml with lichenan and 1.3 mg/ml with laminarin, pH 6.0-7.0, 45°C, wild-type catalytic enzyme domain, kinetic properties of mutant hybrid glucanases, overview Thermotoga maritima
3.2.1.73 additional information
-
 additional information 5.3 mg/ml for mutant TFsW203F with lichenan, pH 6.0-7.0, 45°C. Kinetic properties of parental TFsW203F and mutant hybrid glucanases, overview Fibrobacter succinogenes

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.39 Thermotoga maritima
-
 gene TmLam
-
3.2.1.39 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589
-
 gene TmLam
-
3.2.1.73 Fibrobacter succinogenes
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.39 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by ncikel affiniyt chromatography Thermotoga maritima
3.2.1.73 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Fibrobacter succinogenes

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.73 626
-
 substrate lichenan, pH 6.0-7.0, 45°C, mutant TFsW203F Fibrobacter succinogenes

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.39 laminarin + H2O the mutant enzyme W203F fused to 1,3-beta-D-glucanase catalytic domain shows a 3.6fold increase in specific activity against laminarin as compared with the 1,3-beta-D-glucanase catalytic domain alone with laminaritriose as the major product Thermotoga maritima laminaritriose + ? major product ?
3.2.1.39 laminarin + H2O the mutant enzyme W203F fused to 1,3-beta-D-glucanase catalytic domain shows a 3.6fold increase in specific activity against laminarin as compared with the 1,3-beta-D-glucanase catalytic domain alone with laminaritriose as the major product Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 laminaritriose + ? major product ?
3.2.1.39 lichenan + H2O
-
 Thermotoga maritima ?
-
 ?
3.2.1.39 lichenan + H2O
-
 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 ?
-
 ?
3.2.1.73 laminarin + H2O
-
 Fibrobacter succinogenes ?
-
 ?
3.2.1.73 lichenan + H2O
-
 Fibrobacter succinogenes ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.39 1,3-beta-D-glucanase
-
 Thermotoga maritima
3.2.1.39 laminarinase
-
 Thermotoga maritima
3.2.1.39 TmLam
-
 Thermotoga maritima
3.2.1.73 1,3-1,4-beta-D-glucanase
-
 Fibrobacter succinogenes
3.2.1.73 Lichenase
-
 Fibrobacter succinogenes

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.39 95
-
-
 Thermotoga maritima
3.2.1.73 45
-
 assay at Fibrobacter succinogenes

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.39 30 90 temperature effects on wild-type and mutant hybrid enzymes, overview Thermotoga maritima
3.2.1.73 30 90 temperature effects on mutant W203F and on W203F mutant bifunctional hybrid enzymes, the latter is more resistant to heat treatment than the parental TFsW203F, overview Fibrobacter succinogenes

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.39 38
-
 lichenan pH 6.0-7.0, 45°C, wild-type catalytic enzyme domain Thermotoga maritima
3.2.1.39 109
-
 Laminarin pH 6.0-7.0, 45°C, wild-type catalytic enzyme domain Thermotoga maritima
3.2.1.73 10100
-
 lichenan pH 6.0-7.0, 45°C, mutant TFsW203F Fibrobacter succinogenes

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.39 6 8
-
 Thermotoga maritima
3.2.1.73 6 7 assay at Fibrobacter succinogenes

General Information

EC Number General Information Comment Organism
3.2.1.39 evolution the thermostable laminarinase is composed of a GH family 16 catalytic domain flanked by two family 4 carbohydrate-binding modules (CBM4-1 and CBM4-2) at each terminus Thermotoga maritima
3.2.1.39 physiological function fibrolytic enzyme which plays an important role in the hydrolysis of polysaccharide components. It hydrolyzes internal beta-1,3-glycosidic linkages in beta-1,3-glucans, which form the main component of the cell wall in yeasts and filamentous fungi and the major structural and storage polysaccharide in marine macroalgae such as Laminaria saccharina Thermotoga maritima
3.2.1.73 additional information Glu56, Asp58 and Glu60 residues located in the active site cavity of the enzyme play key roles in enzyme catalysis, functioning as general acid-base residues, structure and functional relationships of Fsbeta-glucanase Fibrobacter succinogenes
3.2.1.73 physiological function fibrolytic enzyme which plays an important role in the hydrolysis of polysaccharide components. It is responsible for precisely hydrolyzing beta-1,4-glycosidic bonds adjacent to the beta-1,3-linkages in lichenan or mixed-linked beta-D-glucans, yielding mainly cellotriose, cellotetraose and cellopentaose Fibrobacter succinogenes