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Literature summary for 1.1.3.9 extracted from

  • Mollerup, F.; Parikka, K.; Vuong, T.V.; Tenkanen, M.; Master, E.
    Influence of a family 29 carbohydrate binding module on the activity of galactose oxidase from Fusarium graminearum (2016), Biochim. Biophys. Acta, 1860, 354-362 .
    View publication on PubMed
Search for more literature for 1.1.3.9

Application

Application Comment Organism
synthesis galactose oxidase (GaO) selectively oxidizes the primary hydroxyl of galactose to a carbonyl, facilitating targeted chemical derivatization of galactose-containing polysaccharides, leading to renewable polymers with tailored physical and chemical properties. Increased substrate binding impeded the action of GaO fusions on more concentrated preparations of galactomannan, galactoglucomannan, and galactoxyloglucan Fusarium graminearum

Cloned(Commentary)

Cloned (Comment) Organism
gene gao, recombinant expression of wild-type enzyme and chimeric enzyme mutants GaO-CBM29 and CBM29-GaO in Pichia pastoris strain KM71H Fusarium graminearum

Protein Variants

Protein Variants Comment Organism
additional information a family 29 glucomannan binding module, CBM29-1-2, from Piromyces equi is separately linked to the N- and C-termini of GaO, effects on enzyme activity and binding of GaO towards various polysaccharides. The chimeric enzyme mutants demonstrate enhanced binding to galactomannan, galactoglucomannan and galactoxyloglucan compared to the wild-type enzyme. The position of the CBM29 fusion affects the enzyme function. Particularly, C-terminal fusion leads to greatest increases in galactomannan binding and catalytic efficiency, where relative to wild-type GaO, kcat/Km values increases by 7.5 and 19.8times on guar galactomannan and locust bean galactomannan, respectively. The fusion of CBM29 also induces oligomerization of GaO–CBM29. Removing CBM32 from wild-type GaO leads to complete loss in enzyme activity, and substituting the native CBM32 for CBM29-1-2 does not regain GaO function Fusarium graminearum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.008
-
 locust bean galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
0.037
-
 guar galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
0.07
-
 galactoxyloglucan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
0.074
-
 galactoglucomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
0.076
-
 galactoxyloglucan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
0.081
-
 guar galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
0.084
-
 locust bean galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
0.13
-
 galactoglucomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
0.14
-
 galactoxyloglucan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
0.16
-
 galactoglucomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
0.19
-
 locust bean galactomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
0.22
-
 guar galactomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
42.4
-
 D-galactose pH 7.0, 30°C, recombinant wild-type enzyme Fusarium graminearum
56
-
 D-galactose pH 7.0, 30°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
60.4
-
 D-galactose pH 7.0, 30°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ a copper oxidase Fusarium graminearum
additional information addition of copper sulfate to shake-flask cultivations doubles mutant GaO-CBM29 activity, whereas enzyme production in a bioreactor system increases the yield of CBM29-GaO and GaO-CBM29 by more than 12times and 6times, respectively. Corresponding specific activities also increase by more than 20%. Addition of 0.5 mM copper (II) sulfate to reaction mixtures containing purified GaO, CBM29-GaO, and GaO-CBM29, or treatment with potassium ferricyanide, does not increase corresponding specific activities, confirming that the purified proteins are in the fully oxidized and active state Fusarium graminearum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-galactose + O2 Fusarium graminearum
-
 D-galacto-hexodialdose + H2O2
-
 ?

Organism

Organism UniProt Comment Textmining
Fusarium graminearum P0CS93 i.e. Gibberella zeae
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type enzyme and chimeric enzyme mutants GaO-CBM29 and CBM29-GaO from Pichia pastoris strain KM71H by nickel affinity and hydrophobic interaction chromatography or vice-versa, follwed by dialysis and anion exchange chromatography Fusarium graminearum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose + O2
-
 Fusarium graminearum D-galacto-hexodialdose + H2O2
-
 ?
galactoglucomannan + O2
-
 Fusarium graminearum ?
-
 ?
galactoxyloglucan + O2
-
 Fusarium graminearum ?
-
 ?
guar galactomannan + O2
-
 Fusarium graminearum ?
-
 ?
locust bean galactomannan + O2
-
 Fusarium graminearum ?
-
 ?
additional information galactose oxidase (GaO) selectively oxidizes the primary hydroxyl of galactose to a carbonyl, facilitating targeted chemical derivatization of galactose-containing polysaccharides, leading to renewable polymers with tailored physical and chemical properties. The activity of wild-type GaO and GaO fusions is measured using the chromogenic ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) assay Fusarium graminearum ?
-
 ?

Synonyms

Synonyms Comment Organism
GAO
-
 Fusarium graminearum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 galactose-containing polysaccharides oxidation assay at Fusarium graminearum
30
-
 galactose oxidation assay at Fusarium graminearum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.25
-
 galactoglucomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
3.33
-
 galactoglucomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
3.37
-
 locust bean galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
3.57
-
 locust bean galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
3.65
-
 galactoxyloglucan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
3.68
-
 galactoxyloglucan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
3.98
-
 galactoglucomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
4.3
-
 locust bean galactomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
4.57
-
 galactoxyloglucan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
5.18
-
 guar galactomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
5.22
-
 guar galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
6.7
-
 guar galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
400
-
 D-galactose pH 7.0, 30°C, recombinant wild-type enzyme Fusarium graminearum
516.7
-
 D-galactose pH 7.0, 30°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
600
-
 D-galactose pH 7.0, 30°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Fusarium graminearum

General Information

General Information Comment Organism
additional information influence of a family 29 carbohydrate binding module on the activity of galactose oxidase from Fusarium graminearum Fusarium graminearum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
9.23
-
 D-galactose pH 7.0, 30°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
9.43
-
 D-galactose pH 7.0, 30°C, recombinant wild-type enzyme Fusarium graminearum
9.93
-
 D-galactose pH 7.0, 30°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
20.31
-
 galactoglucomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
22.63
-
 locust bean galactomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
23.02
-
 galactoxyloglucan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
23.56
-
 guar galactomannan pH 7.0, 25°C, recombinant wild-type enzyme Fusarium graminearum
26.07
-
 galactoxyloglucan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
30.64
-
 galactoglucomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
45.05
-
 galactoglucomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
60.09
-
 galactoxyloglucan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
64.4
-
 guar galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
75.79
-
 locust bean galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant CBM29-GaO Fusarium graminearum
181.08
-
 guar galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum
445.8
-
 locust bean galactomannan pH 7.0, 25°C, recombinant chimeric enzyme mutant GaO-CBM29 Fusarium graminearum