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Literature summary for 2.4.1.30 extracted from

  • De Doncker, M.; Vleminckx, S.; Franceus, J.; Vercauteren, R.; Desmet, T.
    Converting bulk sugars into functional fibers discovery and application of a thermostable beta-1,3-oligoglucan phosphorylase (2024), J. Agric. Food Chem., 72, 10497-10505.
    View publication on PubMed
Search for more literature for 2.4.1.30

Application

Application Comment Organism
synthesis glycoside phosphorylases are promising catalysts for the bottom-up synthesis of beta-1,3-(oligo)glucans since they combine strict regioselectivity with a cheap donor substrate (i.e., alpha-glucose 1-phosphate). But the need for an expensive priming substrate (e.g., laminaribiose) and the tendency to produce shorter oligosaccharides still major bottlenecks. The thermostable beta-1,3-oligoglucan phosphorylase can be useful in converting bulk sugars into functional fibers. Coupling of enzye AtbetaOGP with a thermostable variant of Bifidobacterium adolescentis sucrose phosphorylase enables the efficient production of tailorable beta-1,3-(oligo)glucans from sucrose, with a near-complete conversion of over 99 mol% Anaerolinea thermophila
synthesis glycoside phosphorylases are promising catalysts for the bottom-up synthesis of beta-1,3-(oligo)glucans since they combine strict regioselectivity with a cheap donor substrate (i.e., alpha-glucose 1-phosphate). But the need for an expensive priming substrate (e.g., laminaribiose) and the tendency to produce shorter oligosaccharides still major bottlenecks. The thermostable beta-1,3-oligoglucan phosphorylase can be useful in converting bulk sugars into functional fibers. Coupling of enzyme AtbetaOGP with a thermostable variant of Bifidobacterium adolescentis sucrose phosphorylase enables the efficient production of tailorable beta-1,3-(oligo)glucans from sucrose, with a near-complete conversion of over 99 mol% Euglena gracilis var. saccharophila

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of His-tagged enzyme in Escherichia coli Anaerolinea thermophila

Protein Variants

Protein Variants Comment Organism
additional information coupling of enzye AtbetaOGP with a thermostable variant of Bifidobacterium adolescentis sucrose phosphorylase enables the efficient production of tailorable beta-1,3-(oligo)glucans from sucrose, with a near-complete conversion of over 99 mol% Anaerolinea thermophila
additional information coupling of enzye AtbetaOGP with a thermostable variant of Bifidobacterium adolescentis sucrose phosphorylase enables the efficient production of tailorable beta-1,3-(oligo)glucans from sucrose, with a near-complete conversion of over 99 mol% Euglena gracilis var. saccharophila

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Anaerolinea thermophila
0.56
-
 D-glucose pH 7.0, 30°C recombinant enzyme Euglena gracilis var. saccharophila
5.8
-
 D-glucose pH 6.5, 50°C recombinant enzyme Anaerolinea thermophila

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
133000
-
 sequence calculation Anaerolinea thermophila

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
[(1->3)-beta-D-glucosyl]n + alpha-D-glucose 1-phosphate Euglena gracilis var. saccharophila
-
 [(1->3)-beta-D-glucosyl]n+1 + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate Euglena gracilis var. saccharophila
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila DSM 14523
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila JCM 11388
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila NBRC 100420
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila UNI-1
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila DSM 14523
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila JCM 11388
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila NBRC 100420
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila UNI-1
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r

Organism

Organism UniProt Comment Textmining
Anaerolinea thermophila E8N5S1
-
-
Anaerolinea thermophila DSM 14523 E8N5S1
-
-
Anaerolinea thermophila JCM 11388 E8N5S1
-
-
Anaerolinea thermophila NBRC 100420 E8N5S1
-
-
Anaerolinea thermophila UNI-1 E8N5S1
-
-
Euglena gracilis var. saccharophila A0A2I6SUQ9 cf. EC 2.4.1.97
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, dialysis, and ultrafiltration Anaerolinea thermophila

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila laminaribiose + phosphate
-
 r
D-glucose + alpha-D-glucose 1-phosphate
-
 Euglena gracilis var. saccharophila laminaribiose + phosphate
-
 r
D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila DSM 14523 laminaribiose + phosphate
-
 r
D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila JCM 11388 laminaribiose + phosphate
-
 r
D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila NBRC 100420 laminaribiose + phosphate
-
 r
D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila UNI-1 laminaribiose + phosphate
-
 r
additional information analysis of product profile. Specific activity of AtbetaOGP toward alternative acceptor substrates sucrose and fructose in a coupled reaction with sucrose phosphorylase (SP), overview. Bifidobacterium SP (BaSP) and couples its activity to that of AtbetaOGP. BaSP converts sucrose into alphaG1P in the presence of inorganic phosphate (Pi), while AtbetaOGP utilizes the formed glucosyl phosphate to iteratively elongate growing beta-1,3-(oligo)glucan chains, thereby releasing Pi and enabling its recycling Anaerolinea thermophila ?
-
-
additional information analysis of product profile. Specific activity of AtbetaOGP toward alternative acceptor substrates sucrose and fructose in a coupled reaction with sucrose phosphorylase (SP), overview. Bifidobacterium SP (BaSP) and couples its activity to that of AtbetaOGP. BaSP converts sucrose into alphaG1P in the presence of inorganic phosphate (Pi), while AtbetaOGP utilizes the formed glucosyl phosphate to iteratively elongate growing beta-1,3-(oligo)glucan chains, thereby releasing Pi and enabling its recycling Anaerolinea thermophila DSM 14523 ?
-
-
additional information analysis of product profile. Specific activity of AtbetaOGP toward alternative acceptor substrates sucrose and fructose in a coupled reaction with sucrose phosphorylase (SP), overview. Bifidobacterium SP (BaSP) and couples its activity to that of AtbetaOGP. BaSP converts sucrose into alphaG1P in the presence of inorganic phosphate (Pi), while AtbetaOGP utilizes the formed glucosyl phosphate to iteratively elongate growing beta-1,3-(oligo)glucan chains, thereby releasing Pi and enabling its recycling Anaerolinea thermophila JCM 11388 ?
-
-
additional information analysis of product profile. Specific activity of AtbetaOGP toward alternative acceptor substrates sucrose and fructose in a coupled reaction with sucrose phosphorylase (SP), overview. Bifidobacterium SP (BaSP) and couples its activity to that of AtbetaOGP. BaSP converts sucrose into alphaG1P in the presence of inorganic phosphate (Pi), while AtbetaOGP utilizes the formed glucosyl phosphate to iteratively elongate growing beta-1,3-(oligo)glucan chains, thereby releasing Pi and enabling its recycling Anaerolinea thermophila NBRC 100420 ?
-
-
additional information analysis of product profile. Specific activity of AtbetaOGP toward alternative acceptor substrates sucrose and fructose in a coupled reaction with sucrose phosphorylase (SP), overview. Bifidobacterium SP (BaSP) and couples its activity to that of AtbetaOGP. BaSP converts sucrose into alphaG1P in the presence of inorganic phosphate (Pi), while AtbetaOGP utilizes the formed glucosyl phosphate to iteratively elongate growing beta-1,3-(oligo)glucan chains, thereby releasing Pi and enabling its recycling Anaerolinea thermophila UNI-1 ?
-
-
[(1->3)-beta-D-glucosyl]n + alpha-D-glucose 1-phosphate
-
 Euglena gracilis var. saccharophila [(1->3)-beta-D-glucosyl]n+1 + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate
-
 Euglena gracilis var. saccharophila [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila DSM 14523 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila JCM 11388 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila NBRC 100420 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila UNI-1 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila DSM 14523 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila JCM 11388 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila NBRC 100420 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
[(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila UNI-1 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r

Synonyms

Synonyms Comment Organism
ANT_17590
-
 Anaerolinea thermophila
ANT_17590
-
 Euglena gracilis var. saccharophila
AtbetaOGP
-
 Anaerolinea thermophila
beta-1,3-oligoglucan phosphorylase
-
 Anaerolinea thermophila
beta-1,3-oligoglucan phosphorylase
-
 Euglena gracilis var. saccharophila
betaOGP
-
 Anaerolinea thermophila
betaOGP
-
 Euglena gracilis var. saccharophila
EgbetaOGP
-
 Euglena gracilis var. saccharophila
additional information cf. EC 2.4.1.97 Euglena gracilis var. saccharophila

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Euglena gracilis var. saccharophila
65
-
-
 Anaerolinea thermophila

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
45 75 activity range, profile overview, inactivation at 80°C Anaerolinea thermophila

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
100
-
 purified enzyme, 10 min, inactivation Anaerolinea thermophila

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1
-
 D-glucose pH 7.0, 30°C recombinant enzyme Euglena gracilis var. saccharophila
784
-
 D-glucose pH 6.5, 50°C recombinant enzyme Anaerolinea thermophila

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5 7
-
 Anaerolinea thermophila
7
-
 assay at Euglena gracilis var. saccharophila

pH Range

pH Minimum pH Maximum Comment Organism
5 8.5 activity range, profile overview, over 80% of maximal activity at pH 6.5-7.5 Anaerolinea thermophila

General Information

General Information Comment Organism
evolution the enzyme belongs to glycoside family 149, GH149 Anaerolinea thermophila
evolution the enzyme belongs to glycoside family 149, GH149 Euglena gracilis var. saccharophila
additional information comparisons of the beta-1,3-(oligo)glucan product profiles of betaOGPs from Anaerolinea thermophila (AtbetaOGP) and Euglena gracilis (EgbetaOGP) Anaerolinea thermophila
additional information comparisons of the beta-1,3-(oligo)glucan product profiles of betaOGPs from Anaerolinea thermophila (AtbetatOGP) and Euglena gracilis (EgbetaOGP) Euglena gracilis var. saccharophila
physiological function thermostable beta-1,3-oligoglucan phosphorylase can convert bulk sugars into functional fibers. The enzyme displays a 20fold higher catalytic efficiency than any betaOGP or betaGP reported to date, and it conveniently synthesizes insoluble beta-1,3-glucans up to DP 75 from glucose as a priming substrate Anaerolinea thermophila
physiological function thermostable beta-1,3-oligoglucan phosphorylase can convert bulk sugars into functional fibers. The enzyme displays a 20fold higher catalytic efficiency than any betaOGP or betaGP reported to date, and it conveniently synthesizes insoluble beta-1,3-glucans up to DP 75 from glucose as a priming substrate Euglena gracilis var. saccharophila

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.99
-
 D-glucose pH 7.0, 30°C recombinant enzyme Euglena gracilis var. saccharophila
135
-
 D-glucose pH 6.5, 50°C recombinant enzyme Anaerolinea thermophila