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Literature summary 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

Application

EC Number Application Comment Organism
2.4.1.30 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
2.4.1.30 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.30 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
2.4.1.30 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.30 additional information
-
 additional information Michaelis-Menten kinetics Anaerolinea thermophila
2.4.1.30 0.56
-
 D-glucose pH 7.0, 30°C recombinant enzyme Euglena gracilis var. saccharophila
2.4.1.30 5.8
-
 D-glucose pH 6.5, 50°C recombinant enzyme Anaerolinea thermophila

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.30 133000
-
 sequence calculation Anaerolinea thermophila

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.30 [(1->3)-beta-D-glucosyl]n + alpha-D-glucose 1-phosphate Euglena gracilis var. saccharophila
-
 [(1->3)-beta-D-glucosyl]n+1 + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate Euglena gracilis var. saccharophila
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila DSM 14523
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila JCM 11388
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila NBRC 100420
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate Anaerolinea thermophila UNI-1
-
 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila DSM 14523
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila JCM 11388
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate Anaerolinea thermophila NBRC 100420
-
 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(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

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

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.30 D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila laminaribiose + phosphate
-
 r
2.4.1.30 D-glucose + alpha-D-glucose 1-phosphate
-
 Euglena gracilis var. saccharophila laminaribiose + phosphate
-
 r
2.4.1.30 D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila DSM 14523 laminaribiose + phosphate
-
 r
2.4.1.30 D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila JCM 11388 laminaribiose + phosphate
-
 r
2.4.1.30 D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila NBRC 100420 laminaribiose + phosphate
-
 r
2.4.1.30 D-glucose + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila UNI-1 laminaribiose + phosphate
-
 r
2.4.1.30 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 ?
-
-
2.4.1.30 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 ?
-
-
2.4.1.30 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 ?
-
-
2.4.1.30 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 ?
-
-
2.4.1.30 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 ?
-
-
2.4.1.30 [(1->3)-beta-D-glucosyl]n + alpha-D-glucose 1-phosphate
-
 Euglena gracilis var. saccharophila [(1->3)-beta-D-glucosyl]n+1 + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate
-
 Euglena gracilis var. saccharophila [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila DSM 14523 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila JCM 11388 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila NBRC 100420 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n + phosphate
-
 Anaerolinea thermophila UNI-1 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila DSM 14523 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila JCM 11388 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(1->3)-beta-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 Anaerolinea thermophila NBRC 100420 [(1->3)-beta-D-glucosyl]n + phosphate
-
 r
2.4.1.30 [(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

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.30 30
-
 assay at Euglena gracilis var. saccharophila
2.4.1.30 65
-
-
 Anaerolinea thermophila

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.1.30 45 75 activity range, profile overview, inactivation at 80°C Anaerolinea thermophila

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.30 100
-
 purified enzyme, 10 min, inactivation Anaerolinea thermophila

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.30 1.1
-
 D-glucose pH 7.0, 30°C recombinant enzyme Euglena gracilis var. saccharophila
2.4.1.30 784
-
 D-glucose pH 6.5, 50°C recombinant enzyme Anaerolinea thermophila

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.30 6.5 7
-
 Anaerolinea thermophila
2.4.1.30 7
-
 assay at Euglena gracilis var. saccharophila

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.30 5 8.5 activity range, profile overview, over 80% of maximal activity at pH 6.5-7.5 Anaerolinea thermophila

General Information

EC Number General Information Comment Organism
2.4.1.30 evolution the enzyme belongs to glycoside family 149, GH149 Anaerolinea thermophila
2.4.1.30 evolution the enzyme belongs to glycoside family 149, GH149 Euglena gracilis var. saccharophila
2.4.1.30 additional information comparisons of the beta-1,3-(oligo)glucan product profiles of betaOGPs from Anaerolinea thermophila (AtbetaOGP) and Euglena gracilis (EgbetaOGP) Anaerolinea thermophila
2.4.1.30 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
2.4.1.30 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
2.4.1.30 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.30 1.99
-
 D-glucose pH 7.0, 30°C recombinant enzyme Euglena gracilis var. saccharophila
2.4.1.30 135
-
 D-glucose pH 6.5, 50°C recombinant enzyme Anaerolinea thermophila