Literature summary for 3.2.1.73 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.

Search for more literature for 3.2.1.73

Cloned(Commentary)

Cloned (Comment)	Organism
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

Protein Variants	Comment	Organism
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

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

Molecular Weight [Da]

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

Organism

Organism	UniProt	Comment	Textmining
Bacillus licheniformis	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
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]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
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)

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

Subunits

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

Synonyms

Synonyms	Comment	Organism
LicA	-	Bacillus licheniformis

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
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
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

General Information	Comment	Organism
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

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Release 2023.1 (January 2023)