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Literature summary for 3.2.1.73 extracted from

  • Bernardi, A.V.; de Gouvea, P.F.; Gerolamo, L.E.; Yonamine, D.K.; de Lourdes de Lima Balico, L.; Uyemura, S.A.; Dinamarco, T.M.
    Functional characterization of GH7 endo-1,4-beta-glucanase from Aspergillus fumigatus and its potential industrial application (2018), Protein Expr. Purif., 150, 1-11 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
2-mercaptoethanol activation to 132.1% at 5 mM Aspergillus fumigatus
DMSO activation to 111.5% at 5 mM Aspergillus fumigatus
DTT activation to 165.2% at 5 mM Aspergillus fumigatus
EDTA activation to 108.1% at 5 mM Aspergillus fumigatus
L-cysteine activation to 152.2% at 5 mM Aspergillus fumigatus
Tween 20 activation to 106.2% at 5 mM Aspergillus fumigatus

Application

Application Comment Organism
additional information upon addition to a commercial cellulase cocktail, Af-EGL7 significantly improves sugarcane exploded bagasse (SEB) saccharification, which suggests that enzyme Af-EGL7 has great potential to hydrolyze complex biomass. Af-EGL7 is a promising candidate to enhance enzyme cocktails used in biorefineries such as consolidated bioprocessing Aspergillus fumigatus

Cloned(Commentary)

Cloned (Comment) Organism
gene AFUA_6G01800 or Af-egl7, DNA and amino acid sequnce determination and analysis, sequence comparisons, quantitative RT-PCR exoression analysis, recombinant expression in Escherichia coli strain Rosetta (DE3) pLysS in inclusion bodies, subcloning in Escherichia coli strain DH10beta Aspergillus fumigatus

Inhibitors

Inhibitors Comment Organism Structure
Ag+ complete inhibition at 5 mM Aspergillus fumigatus
Cu2+ 32.4% inhibition at 5 mM Aspergillus fumigatus
Mn2+ activates by 233% at 5 mM, best at 10 mM, slightly inhibitory at 20 mM Aspergillus fumigatus
SDS complete inhibition Aspergillus fumigatus
Zn2+ 20.4% inhibition at 5 mM Aspergillus fumigatus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Km for carboxymethyl cellulose is 209.7 mg/ml and for barley beta-glucan 113.9 mg/ml in presence of 5 mM Mn2+ Aspergillus fumigatus

Localization

Localization Comment Organism GeneOntology No. Textmining
extracellular N-terminal signal peptide at residues 1-20 (MDSKRGVVAAVLALLPLVSA), enzyme Af-EGL7 is secreted Aspergillus fumigatus
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
(NH4)2SO4 activation to 105.5% at 5 mM Aspergillus fumigatus
Ca2+ activation to 135.7% at 5 mM Aspergillus fumigatus
Co2+ activation to 161.1% at 5 mM Aspergillus fumigatus
Fe2+ activation to 128.1% at 5 mM Aspergillus fumigatus
KCl activation to 110.4% at 5 mM Aspergillus fumigatus
Mg2+ activation to 122.6% at 5 mM Aspergillus fumigatus
Mn2+ activates by 233%, best at 10 mM, slightly inhibitory at 20 mM Aspergillus fumigatus
NaCl activation to 113.2% at 5 mM Aspergillus fumigatus
Ni2+ activation to 114.2% at 5 mM Aspergillus fumigatus

Organism

Organism UniProt Comment Textmining
Aspergillus fumigatus Q4WCM9 i.e. Aspergillus fumigatus
-
Aspergillus fumigatus Af293 Q4WCM9 i.e. Aspergillus fumigatus
-
Aspergillus fumigatus ATCC MYA-4609 Q4WCM9 i.e. Aspergillus fumigatus
-
Aspergillus fumigatus CBS 101355 Q4WCM9 i.e. Aspergillus fumigatus
-
Aspergillus fumigatus FGSC A1100 Q4WCM9 i.e. Aspergillus fumigatus
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein the enzyme has four potential N-glycosylation sites (N96, N201, N205, and N379) that are highly conserved among GH7 endoglucanases from other Aspergillus species Aspergillus fumigatus

Renatured (Commentary)

Renatured (Comment) Organism
recombinant enzyme from inclusion bodies by denaturing and dialysis against Tris 10 mM, TCEP 0.1 mM, 0.05 mM EDTA, and 0.06% v/v N-lauroylsarcosine, pH 8.0, and protein refolding in 50 mM Tris, 12.5 mM cyclodextrin, and 1 mM EDTA, pH 7.5 by dialysis Aspergillus fumigatus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
barley beta-glucan + H2O best substrate, 1% w/v substrate concentration, best substrate Aspergillus fumigatus ?
-
?
barley beta-glucan + H2O best substrate, 1% w/v substrate concentration, best substrate Aspergillus fumigatus CBS 101355 ?
-
?
barley beta-glucan + H2O best substrate, 1% w/v substrate concentration, best substrate Aspergillus fumigatus ATCC MYA-4609 ?
-
?
barley beta-glucan + H2O best substrate, 1% w/v substrate concentration, best substrate Aspergillus fumigatus FGSC A1100 ?
-
?
barley beta-glucan + H2O best substrate, 1% w/v substrate concentration, best substrate Aspergillus fumigatus Af293 ?
-
?
carboxymethyl cellulose + H2O 1% w/v substrate concentration, low activity Aspergillus fumigatus ?
-
?
carboxymethyl cellulose + H2O 1% w/v substrate concentration, low activity Aspergillus fumigatus CBS 101355 ?
-
?
carboxymethyl cellulose + H2O 1% w/v substrate concentration, low activity Aspergillus fumigatus ATCC MYA-4609 ?
-
?
carboxymethyl cellulose + H2O 1% w/v substrate concentration, low activity Aspergillus fumigatus FGSC A1100 ?
-
?
carboxymethyl cellulose + H2O 1% w/v substrate concentration, low activity Aspergillus fumigatus Af293 ?
-
?
additional information the enzyme exhibits higher activity toward beta-glucan than toward xyloglucan and carboxymethyl cellulose suggesting that the enzyme corresponds to a beta-1,3-1,4-glucanase. No activity with Avicel and beechwood xylan Aspergillus fumigatus ?
-
?
additional information the enzyme exhibits higher activity toward beta-glucan than toward xyloglucan and carboxymethyl cellulose suggesting that the enzyme corresponds to a beta-1,3-1,4-glucanase. No activity with Avicel and beechwood xylan Aspergillus fumigatus CBS 101355 ?
-
?
additional information the enzyme exhibits higher activity toward beta-glucan than toward xyloglucan and carboxymethyl cellulose suggesting that the enzyme corresponds to a beta-1,3-1,4-glucanase. No activity with Avicel and beechwood xylan Aspergillus fumigatus ATCC MYA-4609 ?
-
?
additional information the enzyme exhibits higher activity toward beta-glucan than toward xyloglucan and carboxymethyl cellulose suggesting that the enzyme corresponds to a beta-1,3-1,4-glucanase. No activity with Avicel and beechwood xylan Aspergillus fumigatus FGSC A1100 ?
-
?
additional information the enzyme exhibits higher activity toward beta-glucan than toward xyloglucan and carboxymethyl cellulose suggesting that the enzyme corresponds to a beta-1,3-1,4-glucanase. No activity with Avicel and beechwood xylan Aspergillus fumigatus Af293 ?
-
?
sugarcane exploded bagasse + H2O 1% w/v substrate concentration, sugarcane exploded bagasse biomass (SEB). SEB is a highly heterogeneous material that consists of 47.5% cellulose, 9.0% hemicellulose, and 34.3% lignin Aspergillus fumigatus ?
-
?
sugarcane exploded bagasse + H2O 1% w/v substrate concentration, sugarcane exploded bagasse biomass (SEB). SEB is a highly heterogeneous material that consists of 47.5% cellulose, 9.0% hemicellulose, and 34.3% lignin Aspergillus fumigatus CBS 101355 ?
-
?
sugarcane exploded bagasse + H2O 1% w/v substrate concentration, sugarcane exploded bagasse biomass (SEB). SEB is a highly heterogeneous material that consists of 47.5% cellulose, 9.0% hemicellulose, and 34.3% lignin Aspergillus fumigatus ATCC MYA-4609 ?
-
?
sugarcane exploded bagasse + H2O 1% w/v substrate concentration, sugarcane exploded bagasse biomass (SEB). SEB is a highly heterogeneous material that consists of 47.5% cellulose, 9.0% hemicellulose, and 34.3% lignin Aspergillus fumigatus FGSC A1100 ?
-
?
sugarcane exploded bagasse + H2O 1% w/v substrate concentration, sugarcane exploded bagasse biomass (SEB). SEB is a highly heterogeneous material that consists of 47.5% cellulose, 9.0% hemicellulose, and 34.3% lignin Aspergillus fumigatus Af293 ?
-
?
xyloglucan + H2O high activity, xyloglucan from tamarind seed, 1% w/v substrate concentration Aspergillus fumigatus ?
-
?
xyloglucan + H2O high activity, xyloglucan from tamarind seed, 1% w/v substrate concentration Aspergillus fumigatus CBS 101355 ?
-
?
xyloglucan + H2O high activity, xyloglucan from tamarind seed, 1% w/v substrate concentration Aspergillus fumigatus ATCC MYA-4609 ?
-
?
xyloglucan + H2O high activity, xyloglucan from tamarind seed, 1% w/v substrate concentration Aspergillus fumigatus FGSC A1100 ?
-
?
xyloglucan + H2O high activity, xyloglucan from tamarind seed, 1% w/v substrate concentration Aspergillus fumigatus Af293 ?
-
?

Subunits

Subunits Comment Organism
? x * 52000, recombinant enzyme, SDS-PAGE, x * 48000, about, sequence calculation Aspergillus fumigatus

Synonyms

Synonyms Comment Organism
Af-EGL7
-
Aspergillus fumigatus
Afu6g01800
-
Aspergillus fumigatus
beta-1,3-1,4-glucanase
-
Aspergillus fumigatus
GH7 endo-1,4-beta-glucanase
-
Aspergillus fumigatus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
Aspergillus fumigatus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 80 40% of maximal activity at 30°C, 80% at 45°C and 60°C, about 35% at 70°C, and 20% at 80°C, profile overview Aspergillus fumigatus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30 60 completely stable up to 50°C for 1 h and loss of 10% activity after 48 h at 50°C, 50% activity remaing at 55°C after 30 min and inactivation after 5 h at 55°C, rapid inactivation at 60°C Aspergillus fumigatus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
43.32
-
carboxymethyl cellulose pH 5.0, 55°C, recombinant enzyme Aspergillus fumigatus
159.9
-
Barley beta-glucan pH 5.0, 55°C, recombinant enzyme Aspergillus fumigatus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
-
Aspergillus fumigatus

pH Range

pH Minimum pH Maximum Comment Organism
3 6.5 activity range, over 50% of maximal activity at pH 4.0-5.5, profile overview Aspergillus fumigatus

pH Stability

pH Stability pH Stability Maximum Comment Organism
3 8 completely stable up to pH 6.0, 40% activity remaining at pH 7.0, inactivation at pH 8.0 Aspergillus fumigatus

Expression

Organism Comment Expression
Aspergillus fumigatus substrate sugarcane exploded bagasse (SEB), induces Af-egl7 by 2500fold up

General Information

General Information Comment Organism
evolution the enzyme belongs to the glycosyl hydrolase family 7, GH7 Aspergillus fumigatus
additional information residues 424-460 from the carbohydrate-binding domain, CBM1 domain Aspergillus fumigatus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
additional information Km for carboxymethyl cellulose is 0.207 mg/ml/s and for barley beta-glucan 1.404 mg/ml/s in presence of 5 mM Mn2+ Aspergillus fumigatus