Cloned (Comment) | Organism |
---|---|
recombinant expression of soluble enzyme chimeric mutant CtGH1-L1-CtGH5-F194A in Escherichia coli strain BL21 | Acetivibrio thermocellus |
Protein Variants | Comment | Organism |
---|---|---|
F194A | site-directed mutagenesis, the mutant shows 2fold increased activity compared to wild-type enzyme | Acetivibrio thermocellus |
additional information | enzyme mutant CtGH5-F194A is fused with a beta-1,4-glucosidase, CtGH1 from Clostridium thermocellum to develop a chimeric enzyme. Improved structural integrity, thermostability and enhanced bifunctional enzyme activities of the chimeric mutant compared to the single point mutant | Acetivibrio thermocellus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics, overview | Acetivibrio thermocellus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetivibrio thermocellus | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
40 | - |
enzyme mutant F194A, pH 5.5, 50°C | Acetivibrio thermocellus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
beta-glucan + H2O | - |
Acetivibrio thermocellus | ? | - |
? | |
carboxymethyl cellulose + H2O | - |
Acetivibrio thermocellus | cellobiose + cellooligosaccharides | - |
? | |
lichenan + H2O | - |
Acetivibrio thermocellus | ? | - |
? | |
additional information | final hydrolysed product produced from carboxymethyl cellulose by chimera 1 is glucose confirming both beta-1,4-endoglucanase and beta-1,4-glucosidase activities, while the products of CtGH5-F194A point mutant are cellobiose and cello-oligosaccharides. Product indentification by thin layer chromatography. Enzymatic hydrolysis of 1% w/v Sorghum stalk pretreated by 1% NaOH by mutant CtGH5-F194A with or without wild-type enzyme CtGH1, which exhibits beta-glucosidase activity. No activity of the wild-type enzyme and mutant F194A with 4-nitrophenyl beta-D-glucoside, which is a substrate of the beta-glucosidase activity of chimeric mutant CtGH1-CtGH5-F194A | Acetivibrio thermocellus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta-1,4-endoglucanase | - |
Acetivibrio thermocellus |
CtGH5 | - |
Acetivibrio thermocellus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
mutant CtGH5 F194A | Acetivibrio thermocellus |
60 | - |
wild-type CtGH5 and chimeric mutant CtGH1-CtGH5 F194A | Acetivibrio thermocellus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
68 | - |
melting temperature of mutant enzyme CtGH5-F194A | Acetivibrio thermocellus |
79 | - |
melting temperature of wild-type enzyme CtGH5 | Acetivibrio thermocellus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.006 | - |
carboxymethyl cellulose | pH 4.2, 60°C, recombinant wild-type enzyme CtGH5 | Acetivibrio thermocellus | |
0.026 | - |
carboxymethyl cellulose | pH 4.2, 60°C, recombinant mutant CtGH5-F194A | Acetivibrio thermocellus | |
0.045 | - |
beta-Glucan | pH 4.2, 60°C, recombinant mutant CtGH1-CtGH5-F194A | Acetivibrio thermocellus | |
0.053 | - |
lichenan | pH 4.2, 60°C, recombinant chimeric mutant CtGH1-CtGH5-F194A | Acetivibrio thermocellus | |
0.09 | - |
beta-Glucan | pH 4.2, 60°C, recombinant wild-type enzyme CtGH5 | Acetivibrio thermocellus | |
0.1 | - |
lichenan | pH 4.2, 60°C, recombinant wild-type enzyme CtGH5 | Acetivibrio thermocellus | |
0.115 | - |
carboxymethyl cellulose | pH 4.2, 60°C, recombinant mutant CtGH1-CtGH5-F194A | Acetivibrio thermocellus | |
0.135 | - |
beta-Glucan | pH 4.2, 60°C, recombinant mutant CtGH5-F194A | Acetivibrio thermocellus | |
0.145 | - |
lichenan | pH 4.2, 60°C, recombinant mutant CtGH5-F194A | Acetivibrio thermocellus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.2 | - |
wild-type CtGH5 | Acetivibrio thermocellus |
5 | - |
chimeric mutant CtGH1-CtGH5 F194A | Acetivibrio thermocellus |
5.5 | - |
mutant CtGH5 F194A | Acetivibrio thermocellus |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycoside hydrolase family 5,GH5 | Acetivibrio thermocellus |