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Literature summary extracted from

  • Lu, H.; Luo, H.; Shi, P.; Huang, H.; Meng, K.; Yang, P.; Yao, B.
    A novel thermophilic endo-beta-1,4-mannanase from Aspergillus nidulans XZ3: functional roles of carbohydrate-binding module and Thr/Ser-rich linker region (2014), Appl. Microbiol. Biotechnol., 98, 2155-2163.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.78 acetone activates the wild-type enzyme and mutant Man5DELTACBM at 40-60% v/v, no effect on mutant Man5DELTACL Aspergillus nidulans
3.2.1.78 SDS the wild-type enzyme slightly at 0.5-1.0% v/v, inhibits the mutant enzymes Aspergillus nidulans
3.2.1.78 Triton X-100 activates the wild-type enzyme slightly at 0.5% v/v, inhibits the mutant enzymes Aspergillus nidulans

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.78 gene man5XZ3, sequence comparisons, cloning in Escherichia coli strain Trans1-T1, and recombinant expression of wild-type and mutant enzymes fused in frame to the DNA sequence for the Saccharomyces cerevisiae alpha-factor secretory signal peptide and under the control of the methanol-inducible alcohol oxidase promoter in Pichia pastoris strain GS115, induction by methanol, the proteins are secreted Aspergillus nidulans

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.78 additional information generation of two truncated enzyme mutants lacking the family 1 carbohydrate-binding module Man5DELTACBM or the family 1 carbohydrate-binding module and the Thr/Ser-rich linker region, Man5DELTACL, respectively. The mutant enzymes show significantly altered secondary structures compared to the wild-type enzyme, overview. Removal of the family 1 carbohydrate-binding module alone improves the thermostability of the enzyme, but additional removal of the linker region results in worse thermostability. The mutants are less stable in presence of acetone, SDS, Triton X-100, or urea compared to the wild-type enzyme Aspergillus nidulans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.78 additional information
-
 additional information Km values with ocust bean gum are 0.9 mg/ml for wild-type enzyme, 0.8 mg/ml for mutant Man5DELTACBM, and 1.1 mg/ml for mutant Man5DELTACL Aspergillus nidulans

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.78 extracellular the enzyme is secreted Aspergillus nidulans
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.78 38000
-
 x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans
3.2.1.78 41000
-
 x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans
3.2.1.78 55000
-
 x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans
3.2.1.78 58000
-
 x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans
3.2.1.78 60000
-
 x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans
3.2.1.78 63000
-
 x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.78 Aspergillus nidulans
-
 gene man5XZ3
-
3.2.1.78 Aspergillus nidulans XZ3
-
 gene man5XZ3
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.2.1.78 glycoprotein one potential N-glycosylation site, Asn98-Phe99-Thr100, the enzyme is deglycosylated by endo-beta-N-acetylglucosaminidase H. The Thr/Ser-rich linker region has numerous putative O-glycosylation sites Aspergillus nidulans

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.78 recombinant wild-type and mutant enzymes from Pichia pastoris strain GS115 culture supernatant by ultrafiltration and anion exchange chromatography to homogeneity Aspergillus nidulans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.78 guar gum + H2O 41.2% of the activity with locust bean gum Aspergillus nidulans ?
-
 ?
3.2.1.78 guar gum + H2O 41.2% of the activity with locust bean gum Aspergillus nidulans XZ3 ?
-
 ?
3.2.1.78 locust bean gum + H2O best substrate Aspergillus nidulans mannobiose + mannotriose + mannotetraose hydrolysis products are 17.2% mannobiose, 37.4% mannotriose, 9.9% mannotetraose, and 35.5% other unidentified oligosaccharides ?
3.2.1.78 locust bean gum + H2O best substrate Aspergillus nidulans XZ3 mannobiose + mannotriose + mannotetraose hydrolysis products are 17.2% mannobiose, 37.4% mannotriose, 9.9% mannotetraose, and 35.5% other unidentified oligosaccharides ?
3.2.1.78 additional information no activity with beechwood xylan, 4-nitrophenyl beta-D-glucopyranoside, 4-nitrophenyl beta-D-mannoside, and carboxymethyl cellulose Aspergillus nidulans ?
-
 ?
3.2.1.78 additional information no activity with beechwood xylan, 4-nitrophenyl beta-D-glucopyranoside, 4-nitrophenyl beta-D-mannoside, and carboxymethyl cellulose Aspergillus nidulans XZ3 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.78 ? x * 63000, glycosylated recombinant wild-type enzyme, SDS-PAGE, x * 58000, glycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 41000, glycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE, x * 60000, deglycosylated recombinant wild-type enzyme, SDS-PAGE, x * 55000, deglycosylated mutant lacking the CBM1 domain, SDS-PAGE, x * 38000, deglycosylated mutant lacking the CBM1 domain and linker region, SDS-PAGE Aspergillus nidulans
3.2.1.78 More the multimodular enzyme consists of a family 1 carbohydrate-binding module (CBM1), a Thr/Ser-rich linker region, and a catalytic domain Aspergillus nidulans

Synonyms

EC Number Synonyms Comment Organism
3.2.1.78 Man5XZ3
-
 Aspergillus nidulans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.78 80
-
-
 Aspergillus nidulans

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.78 30 90 activity range, wild-type enzyme Aspergillus nidulans

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.78 additional information
-
 removal of the family 1 carbohydrate-binding module alone improves the thermostability of the enzyme, but additional removal of the linker region results in worse thermostability Aspergillus nidulans
3.2.1.78 30 90 purified recombinant wild-type enzyme, pH 5.0, 1 h, more than 45% of maximal activity at 50-80°C and over 30% activity at 90°C Aspergillus nidulans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.78 5
-
-
 Aspergillus nidulans

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.78 4 9 activity range, wild-type enzyme, optimum at pH 5.0, 45% of maximal activity at pH 9.0 Aspergillus nidulans

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.78 1 2 purified recombinant wild-type enzyme, 37°C, 1 h, 50% activity remaining Aspergillus nidulans
3.2.1.78 3
-
 purified recombinant wild-type enzyme, 37°C, 1 h, 50% activity remaining Aspergillus nidulans
3.2.1.78 4 9 purified recombinant wild-type enzyme, 37°C, 1 h, stable at Aspergillus nidulans

General Information

EC Number General Information Comment Organism
3.2.1.78 evolution the enzyme belongs to the glycosyl hydrolase family 5, GH5 Aspergillus nidulans