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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces sp. G12 | I7L8N7 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 Codon Plus (DE3) RP | Streptomyces sp. G12 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
carboxymethyl cellulose + H2O | AZO-carboxymethyl cellulose | Streptomyces sp. G12 | ? | - |
? | |
additional information | bioconversion of the pretreated Arundo donax lignocellulosic biomass by wild-type and mutant enzymes | Streptomyces sp. G12 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta-1,4-endoglucanase | - |
Streptomyces sp. G12 |
CelStrep | - |
Streptomyces sp. G12 |
endo-beta-1,4-glucanase | - |
Streptomyces sp. G12 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
recombinant wild-type and mutant enzymes | Streptomyces sp. G12 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
recombinant wild-type and mutant enzymes, assay at | Streptomyces sp. G12 |
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 |