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Literature summary for 3.2.1.176 extracted from

  • Dotsenko, A.S.; Gusakov, A.V.; Volkov, P.V.; Rozhkova, A.M.; Sinitsyn, A.P.
    N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity (2016), Biotechnol. Bioeng., 113, 283-291.
    View publication on PubMed
Search for more literature for 3.2.1.176

Cloned(Commentary)

Cloned (Comment) Organism
expression in Penicillium canescens PCA10 Talaromyces verruculosus

Crystallization (Commentary)

Crystallization (Comment) Organism
3D structural modeling of the Cel7A catalytic module with N-linked glucans attached to Asn45, Asn194, and Asn388 glycosylation sites. Residue Asn45 is located near the entrance to the enzyme active site tunnel, while the two other N-glycosylation sites are located on the loops forming the tunnel, Asn194 being closer to the entrance and Asn388 being closer to the exit of the tunnel Talaromyces verruculosus

Protein Variants

Protein Variants Comment Organism
N194A mutation in glycosylation site, has no notable effect on pH-optimum of activity and enzyme thermostability, leads to decrease in substrate digestion rates Talaromyces verruculosus
N388A mutation in glycosylation site, has no notable effect on pH-optimum of activity and enzyme thermostability, leads to decrease in substrate digestion rates Talaromyces verruculosus
N430A expression of N430A mutant could not be achieved Talaromyces verruculosus
N45A mutation in glycosylation site, has no notable effect on pH-optimum of activity and enzyme thermostability, but leads to a significant increase in the rate of avicel and milled aspen wood hydrolysis Talaromyces verruculosus

Organism

Organism UniProt Comment Textmining
Talaromyces verruculosus
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein sequence shows four potential N-glycosylation sites at the catalytic module: Asn45, Asn194, Asn388, and Asn430. The N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13(GlcNAc)2, or a single GlcNAc residue. Mutations in the glycosylation sites have no notable effect on pH-optimum of activity and enzyme thermostability but influence both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates Talaromyces verruculosus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.028
-
 substrate 4-nitrophenyl beta-D-lactoside, wild-type, pH 5, 50°C Talaromyces verruculosus
0.036
-
 substrate 4-nitrophenyl beta-D-lactoside, mutant N194A, pH 5, 50°C Talaromyces verruculosus
0.038
-
 substrate 4-nitrophenyl beta-D-lactoside, mutant N45A, pH 5, 50°C Talaromyces verruculosus
0.039
-
 substrate 4-nitrophenyl beta-D-lactoside, mutant N388A, pH 5, 50°C Talaromyces verruculosus
0.041
-
 substrate beta-glucan, mutant N388A, pH 5, 50°C Talaromyces verruculosus
0.044
-
 substrate beta-glucan, mutant N194A, pH 5, 50°C Talaromyces verruculosus
0.047
-
 substrate beta-glucan, wild-type, pH 5, 50°C Talaromyces verruculosus
0.117
-
 substrate beta-glucan, mutant N45A, pH 5, 50°C Talaromyces verruculosus
0.17
-
 substrate avicel, mutant N194A, pH 5, 40°C Talaromyces verruculosus
0.19
-
 substrate avicel, mutant N388A, pH 5, 40°C Talaromyces verruculosus
0.2
-
 substrate avicel, wild-type, pH 5, 40°C Talaromyces verruculosus
0.22
-
 substrate avicel, mutant N45A, pH 5, 40°C Talaromyces verruculosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-D-lactoside + H2O
-
 Talaromyces verruculosus 4-nitrophenol + D-lactose
-
 ?
avicel + H2O
-
 Talaromyces verruculosus cellobiose
-
 ?
beta-glucan + H2O
-
 Talaromyces verruculosus ?
-
 ?
additional information no substrates: carboxymethyl cellulose, 4-nitrophenyl-beta-D-glucopyranoside and 4-nitrophenyl-beta-D-galactopyranoside Talaromyces verruculosus ?
-
 ?

Synonyms

Synonyms Comment Organism
Cel7A
-
 Talaromyces verruculosus