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Literature summary extracted from

  • Nath, P.; Dhillon, A.; Kumar, K.; Sharma, K.; Jamaldheen, S.B.; Moholkar, V.S.; Goyal, A.
    Development of bi-functional chimeric enzyme (CtGH1-L1-CtGH5-F194A) from endoglucanase (CtGH5) mutant F194A and ?-1,4-glucosidase (CtGH1) from Clostridium thermocellum with enhanced activity and structural integrity (2019), Biores. Technol., 282, 494-501 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.4 recombinant expression of soluble enzyme chimeric mutant CtGH1-L1-CtGH5-F194A in Escherichia coli strain BL21 Acetivibrio thermocellus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.4 F194A site-directed mutagenesis, the mutant shows 2fold increased activity compared to wild-type enzyme Acetivibrio thermocellus
3.2.1.4 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.4 additional information
-
 additional information Michaelis-Menten kinetics, overview Acetivibrio thermocellus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.4 Acetivibrio thermocellus
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.4 40
-
 enzyme mutant F194A, pH 5.5, 50°C Acetivibrio thermocellus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.4 beta-glucan + H2O
-
 Acetivibrio thermocellus ?
-
 ?
3.2.1.4 carboxymethyl cellulose + H2O
-
 Acetivibrio thermocellus cellobiose + cellooligosaccharides
-
 ?
3.2.1.4 lichenan + H2O
-
 Acetivibrio thermocellus ?
-
 ?
3.2.1.4 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

EC Number Synonyms Comment Organism
3.2.1.4 beta-1,4-endoglucanase
-
 Acetivibrio thermocellus
3.2.1.4 CtGH5
-
 Acetivibrio thermocellus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.4 50
-
 mutant CtGH5 F194A Acetivibrio thermocellus
3.2.1.4 60
-
 wild-type CtGH5 and chimeric mutant CtGH1-CtGH5 F194A Acetivibrio thermocellus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.4 68
-
 melting temperature of mutant enzyme CtGH5-F194A Acetivibrio thermocellus
3.2.1.4 79
-
 melting temperature of wild-type enzyme CtGH5 Acetivibrio thermocellus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.4 0.006
-
 carboxymethyl cellulose pH 4.2, 60°C, recombinant wild-type enzyme CtGH5 Acetivibrio thermocellus
3.2.1.4 0.026
-
 carboxymethyl cellulose pH 4.2, 60°C, recombinant mutant CtGH5-F194A Acetivibrio thermocellus
3.2.1.4 0.045
-
 beta-Glucan pH 4.2, 60°C, recombinant mutant CtGH1-CtGH5-F194A Acetivibrio thermocellus
3.2.1.4 0.053
-
 lichenan pH 4.2, 60°C, recombinant chimeric mutant CtGH1-CtGH5-F194A Acetivibrio thermocellus
3.2.1.4 0.09
-
 beta-Glucan pH 4.2, 60°C, recombinant wild-type enzyme CtGH5 Acetivibrio thermocellus
3.2.1.4 0.1
-
 lichenan pH 4.2, 60°C, recombinant wild-type enzyme CtGH5 Acetivibrio thermocellus
3.2.1.4 0.115
-
 carboxymethyl cellulose pH 4.2, 60°C, recombinant mutant CtGH1-CtGH5-F194A Acetivibrio thermocellus
3.2.1.4 0.135
-
 beta-Glucan pH 4.2, 60°C, recombinant mutant CtGH5-F194A Acetivibrio thermocellus
3.2.1.4 0.145
-
 lichenan pH 4.2, 60°C, recombinant mutant CtGH5-F194A Acetivibrio thermocellus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.4 4.2
-
 wild-type CtGH5 Acetivibrio thermocellus
3.2.1.4 5
-
 chimeric mutant CtGH1-CtGH5 F194A Acetivibrio thermocellus
3.2.1.4 5.5
-
 mutant CtGH5 F194A Acetivibrio thermocellus

General Information

EC Number General Information Comment Organism
3.2.1.4 evolution the enzyme belongs to the glycoside hydrolase family 5,GH5 Acetivibrio thermocellus