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Literature summary for 3.2.1.4 extracted from

  • Cecchini, D.A.; Pepe, O.; Pennacchio, A.; Fagnano, M.; Faraco, V.
    Directed evolution of the bacterial endo-beta-1,4-glucanase from Streptomyces sp. G12 towards improved catalysts for lignocellulose conversion (2018), AMB Express, 8, 74 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.4

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21 CodonPlus (DE3) RP, subcloning in Escherichia coli strain TOP10 Streptomyces sp. G12

Protein Variants

Protein Variants Comment Organism
G145D/N207K random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme Streptomyces sp. G12
G263C/R307H random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme Streptomyces sp. G12
additional information development of biocatalysts for enhanced hydrolysis of (hemi)cellulose into monosaccharides with random diversity by directed evolution of the gene coding for bacterial endo-beta-1,4-glucanase from Streptomyces sp. G12, improved catalysis of lignocellulose conversion, screening of a library of 10000 random mutants, detection of variants with higher activity than the wild-type enzyme, and structure-function relationships of the mutants, overview. Mutations T67N, D142E, T157I, and S218N are located in the catalytic module and mutations G251D, V259D, V242D, and D330E in the carbohydrate binding module (CBM) Streptomyces sp. G12
P228R random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme Streptomyces sp. G12
T157I/G251D/V259D random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme Streptomyces sp. G12
T67N/D142E/S218N/V242D/D330E random mutagenesis, the mutant shows increased activity with carboxymethyl cellulose compared to the wild-type enzyme Streptomyces sp. G12

Organism

Organism UniProt Comment Textmining
Streptomyces sp. G12 I7L8N7
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-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 Codon Plus (DE3) RP Streptomyces sp. G12

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
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 substrate carboxymethyl cellulose, recombinant wild-type enzyme shows 70.4 mU/ml activity, activities of enzyme mutants G263C/R307H, G145D/N207K, P228R, T67N/D142E/S218N/V242D/D330E, and T157I/G251D/V259D are 103.8, 88.6, 112.6, 89.1, and 102.9 mU/ml, respectively Streptomyces sp. G12

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
carboxymethyl cellulose + H2O AZO-carboxymethyl cellulose Streptomyces sp. G12 ?
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 ?
additional information bioconversion of the pretreated Arundo donax lignocellulosic biomass by wild-type and mutant enzymes Streptomyces sp. G12 ?
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 ?

Synonyms

Synonyms Comment Organism
beta-1,4-endoglucanase
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 Streptomyces sp. G12
CelStrep
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 Streptomyces sp. G12
endo-beta-1,4-glucanase
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 Streptomyces sp. G12

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
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 recombinant wild-type and mutant enzymes Streptomyces sp. G12

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 recombinant wild-type and mutant enzymes, assay at Streptomyces sp. G12

General Information

General Information Comment Organism
evolution the enzyme belongs to the glycosyl hydrolase family 12, GH12 Streptomyces sp. G12
additional information homology modeling of the three-dimensional structures of the catalytic and binding modules of recombinant enzyme rCelStrep, overview Streptomyces sp. G12