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Literature summary extracted from

  • van Lieshout, J.F.; Perez Gutierrez, O.N.; Vroom, W.; Planas, A.; de Vos, W.M.; van der Oost, J.; Koutsopoulos, S.
    Thermal stabilization of an endoglucanase by cyclization (2012), Appl. Biochem. Biotechnol., 167, 2039-2053.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.73 expression of the N-terminally His-tagged enzyme with or without inteins, constructs for linear and circular enzymes, in Escherichia coli strains BL21(DE3) or JM109(DE3). The inteins are spliced out Bacillus licheniformis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.73 additional information covalent linkage between the N- and C-termini of a polypeptide chain to create circular variants of the enzyme by an intein-driven protein splicing approach, method, overview. Two circular variants, LicAC1 and LicA-C2, which have connecting loops of 20 and 14 amino acids, respectively, show catalytic activities that are approximately two and three times higher, respectively, compared to that of the linear LicA, LicA-L1. Also the thermal stability of the circular variants is significantly increased compared to the linear form. The circular proteins contain a thrombin recognition site and can be linearized by cleavage with thrombin Bacillus licheniformis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.73 additional information the wild-type enzyme contains a signal peptide Bacillus licheniformis
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.73 27000
-
 1 * 27000, recombinant mature circular enzyme variant LicA-C1 without intein sequences, SDS-PAGE Bacillus licheniformis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.73 Bacillus licheniformis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.73 recombinant N-terminally His-tagged linear and circular enzymes from Escherichia coli strains BL21(DE3) or JM109(DE3) by nickel affinity chromatography Bacillus licheniformis

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 2.7
-
 purified recombinant linear enzyme and wild-type enzyme, substrate barley-beta-glucan, pH and temperature not specified in the publication Bacillus licheniformis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.73 barley-beta-glucan + H2O
-
 Bacillus licheniformis ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.73 monomer 1 * 27000, recombinant mature circular enzyme variant LicA-C1 without intein sequences, SDS-PAGE Bacillus licheniformis

Synonyms

EC Number Synonyms Comment Organism
3.2.1.73 LicA
-
 Bacillus licheniformis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.73 additional information
-
 addition of 1 mM EDTA causes the linear enzyme to denature at 59.8°C, which is lower compared to the melting temperature of 62.6°C in the presence of calcium. At 1 mM EDTA, the circular enzyme variant LicA-C1 shows a melting temperature of 62.8°C, at 1 mM CaCl2 the melting temperature is 66.4°C Bacillus licheniformis
3.2.1.73 65
-
 the linear enzyme form loses 50% activity within 3 min, whereas the two circular variants have 6fold (LicA-C1) and 16fold (LicA-C2) increased half-life time of inactivation Bacillus licheniformis

General Information

EC Number General Information Comment Organism
3.2.1.73 additional information both the loss of the signal peptide (29 amino acids) and the addition of the His-tag to the peptide backbone do not affect the specific activity of linear enzyme Bacillus licheniformis