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Literature summary for 3.2.1.78 extracted from

  • Rosengren, A.; Hägglund, P.; Anderson, L.; Pavon-Orozco, P.; Peterson-Wulff, R.; Nerinckx, W.; Stalbrand, H.
    The role of subsite 2 of the Trichoderma reesei beta-mannanase TrMan5A in hydrolysis and transglycosylation (2012), Biocatal. Biotransform., 30, 338-352 .
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzyme in Pichia pastoris Trichoderma reesei

Protein Variants

Protein Variants Comment Organism
R171K the mutant displays retained activity on polymeric galactomannan but reduced activity on oligosaccharides due to an increase of Km. While the wild-type enzyme produces mannobiose as dominant product from mannotetraose, the R171K mutant enzyme produces mannotriose and mannose. The preferred productive binding mode of mannotetraose is shifted from subsite -2 to +2 in the wild-type to subsite -3 to +1 in the R171K mutant. The wild-type enzyme can perform transglycosylation on to saccharide acceptors while the R171K mutant cannot. Wild-type and mutant enzyme show the ability to perform alcoholysis reactions with methanol and butanol, forming new beta-linked glycoconjugates. It appears that the wild-type enzyme produces mainly mannobiose conjugates using mannotetetraoses substrate, while in contrast the R171K mutant produces mainly mannotriose conjugates, due to the altered subsite binding Trichoderma reesei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
mannohexaose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
0.08
-
Mannopentaose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
0.25
-
4-methylumbelliferyl mannotrioside pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
0.31
-
Mannotetraose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
0.34
-
mannohexaose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
0.46
-
4-methylumbelliferyl mannotrioside pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
0.6
-
locust bean gum pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
1.3
-
locust bean gum pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
1.7
-
Mannopentaose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
10.6
-
Mannotetraose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei

Organism

Organism UniProt Comment Textmining
Trichoderma reesei Q99036
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl mannotrioside + H2O
-
Trichoderma reesei 4-methylumbelliferone + mannotriose
-
?
locust bean gum + H2O
-
Trichoderma reesei ?
-
?
mannohexaose + H2O
-
Trichoderma reesei ?
-
?
mannopentaose + H2O
-
Trichoderma reesei ?
-
?
mannotetraose + H2O the wild-type enzyme produces mannobiose as dominant product from mannotetraose. The R171K mutant enzyme produces mannotriose and mannose Trichoderma reesei ?
-
?
additional information the wild-type enzyme can perform transglycosylation on to saccharide acceptors while the R171K mutant cannot Wild-type and mutant enzyme show the ability to perform alcoholysis reactions with methanol and butanol, forming new beta-linked glycoconjugates. It appears that the wild-type enzyme produces mainly mannobiose conjugates using mannotetetraoses substrate, while in contrast the R171K mutant produces mainly mannotriose conjugates, due to the altered subsite binding Trichoderma reesei ?
-
?

Synonyms

Synonyms Comment Organism
endo-1,4-beta-D-mannanase
-
Trichoderma reesei
TrMan5A
-
Trichoderma reesei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
4-methylumbelliferyl mannotrioside pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
0.03
-
4-methylumbelliferyl mannotrioside pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
2 8 mannohexaose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
3
-
Mannotetraose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
6
-
Mannotetraose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
11
-
Mannopentaose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
13
-
Mannopentaose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
20
-
mannohexaose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
240
-
locust bean gum pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
260
-
locust bean gum pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.02
-
4-methylumbelliferyl mannotrioside pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
0.12
-
4-methylumbelliferyl mannotrioside pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
0.28
-
Mannotetraose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
6.5
-
Mannopentaose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
19.4
-
Mannotetraose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
82.4
-
mannohexaose pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
163
-
Mannopentaose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
200
-
locust bean gum pH 40°C, pH 4.5, mutant enzyme R171K Trichoderma reesei
400
-
locust bean gum pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei
400
-
mannohexaose pH 40°C, pH 4.5, wild-type enzyme Trichoderma reesei