Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Franceus, J.; Decuyper, L.; Dhooghe, M.; Desmet, T.
    Exploring the sequence diversity in glycoside hydrolase family 13_18 reveals a novel glucosylglycerol phosphorylase (2018), Appl. Microbiol. Biotechnol., 102, 3183-3191 .
    View publication on PubMed

Application

EC Number Application Comment Organism
2.4.1.359 synthesis glucosylglycerol phosphorylase might be an attractive biocatalyst for the production of the osmolyte glucosylglycerol, which is currently produced on industrial scale by exploiting a side activity of the closely related sucrose phosphorylase Marinobacter adhaerens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.359 expression in Escherichia coli Marinobacter adhaerens
2.4.1.359 gene gtfA, sequence comparisons and phylogenetic tree, recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Marinobacter adhaerens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.359 homology modeling of structure. Residues Tyr194 and Gln336 each form a hydrogen bond with a hydroxyl group of glycerol, while Ala333 is situated at a suitable location for hydrophobic interaction with atom C1 Marinobacter adhaerens

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.359 A333D site-directed mutagenesis Marinobacter adhaerens
2.4.1.359 Q336A site-directed mutagenesis Marinobacter adhaerens
2.4.1.359 Q336A 2.6% residual activity Marinobacter adhaerens
2.4.1.359 Y194A 2.7% residual activity Marinobacter adhaerens
2.4.1.359 Y196A site-directed mutagenesis Marinobacter adhaerens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.359 additional information
-
 additional information Michaelis-Menten kinetics Marinobacter adhaerens
2.4.1.359 2.5
-
 alpha-D-glucose 1-phosphate pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 2.5
-
 alpha-D-glucose 1-phosphate recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 3.9
-
 2-O-(alpha-D-glucopyranosyl)glycerol pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 3.9
-
 2-O-(alpha-D-glucopyranosyl)glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 7
-
 glycerol pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 7
-
 glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 10
-
 phosphate pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 10
-
 2-O-(alpha-D-glucopyranosyl)glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.359 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate Marinobacter adhaerens
-
 alpha-D-glucose 1-phosphate + glycerol
-
 r
2.4.1.359 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate Marinobacter adhaerens DSM 23420
-
 alpha-D-glucose 1-phosphate + glycerol
-
 r
2.4.1.359 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate Marinobacter adhaerens HP15
-
 alpha-D-glucose 1-phosphate + glycerol
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.359 Marinobacter adhaerens E4PMA5
-
-
2.4.1.359 Marinobacter adhaerens E4PMA5 member of family GH13_18
-
2.4.1.359 Marinobacter adhaerens DSM 23420 E4PMA5
-
-
2.4.1.359 Marinobacter adhaerens DSM 23420 E4PMA5 member of family GH13_18
-
2.4.1.359 Marinobacter adhaerens HP15 E4PMA5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.359 ecombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration Marinobacter adhaerens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.359 12.7
-
 purified recombinant enzyme, pH 6.5, 37°C, phosphorolytic activity Marinobacter adhaerens
2.4.1.359 14.5
-
 pH 6.5, 37°C, acceptor substrate 1,2-propanediol Marinobacter adhaerens
2.4.1.359 17
-
 substrate 2-O-(alpha-D-glucopyranosyl)glycerol, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 33
-
 pH 6.5, 37°C, acceptor substrate glycerol Marinobacter adhaerens
2.4.1.359 33
-
 purified recombinant wild-type enzyme, pH 6.5, 30°C Marinobacter adhaerens
2.4.1.359 45.2
-
 purified recombinant enzyme, pH 6.5, 37°C, synthetic activity Marinobacter adhaerens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.359 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate
-
 Marinobacter adhaerens alpha-D-glucose 1-phosphate + glycerol
-
 r
2.4.1.359 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate
-
 Marinobacter adhaerens DSM 23420 alpha-D-glucose 1-phosphate + glycerol
-
 r
2.4.1.359 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate
-
 Marinobacter adhaerens HP15 alpha-D-glucose 1-phosphate + glycerol
-
 r
2.4.1.359 alpha-D-glucose 1-phosphate + 1,2-propanediol
-
 Marinobacter adhaerens 2-O-(alpha-D-glucopyranosyl)propanediol + phosphate
-
 r
2.4.1.359 alpha-D-glucose 1-phosphate + 1,2-propanediol
-
 Marinobacter adhaerens DSM 23420 2-O-(alpha-D-glucopyranosyl)propanediol + phosphate
-
 r
2.4.1.359 alpha-D-glucose 1-phosphate + 1,2-propanediol
-
 Marinobacter adhaerens HP15 2-O-(alpha-D-glucopyranosyl)propanediol + phosphate
-
 r
2.4.1.359 alpha-D-glucose 1-phosphate + glycerol
-
 Marinobacter adhaerens 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate
-
 r
2.4.1.359 alpha-D-glucose 1-phosphate + glycerol
-
 Marinobacter adhaerens DSM 23420 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate
-
 r
2.4.1.359 alpha-D-glucose 1-phosphate + glycerol
-
 Marinobacter adhaerens HP15 2-O-(alpha-D-glucopyranosyl)glycerol + phosphate
-
 r
2.4.1.359 additional information no transglucosylation activity with fructose, fructose 6-phosphate or glycerate as acceptors Marinobacter adhaerens ?
-
 ?
2.4.1.359 additional information the enzyme catalyses the reversible phosphorolysis of 2-O-alpha-D-glucosylglycerol with retention of the anomeric configuration. Homology modelling, docking and mutagenesis are performed to pinpoint particular acceptor site residues (Tyr194, Ala333, Gln336) involved in the binding of glycerol. In the phosphorolytic direction, the enzyme is active on glucosylglycerol but not on sucrose or glucosylglycerate. A variety of other acceptors are surveyed as well, but significant activity can only be observed using 1,2-propanediol. No transglucosylation activity is detected when fructose, fructose 6-phosphate, or glycerate are supplied as acceptor Marinobacter adhaerens ?
-
-
2.4.1.359 additional information no transglucosylation activity with fructose, fructose 6-phosphate or glycerate as acceptors Marinobacter adhaerens DSM 23420 ?
-
 ?
2.4.1.359 additional information the enzyme catalyses the reversible phosphorolysis of 2-O-alpha-D-glucosylglycerol with retention of the anomeric configuration. Homology modelling, docking and mutagenesis are performed to pinpoint particular acceptor site residues (Tyr194, Ala333, Gln336) involved in the binding of glycerol. In the phosphorolytic direction, the enzyme is active on glucosylglycerol but not on sucrose or glucosylglycerate. A variety of other acceptors are surveyed as well, but significant activity can only be observed using 1,2-propanediol. No transglucosylation activity is detected when fructose, fructose 6-phosphate, or glycerate are supplied as acceptor Marinobacter adhaerens DSM 23420 ?
-
-
2.4.1.359 additional information no transglucosylation activity with fructose, fructose 6-phosphate or glycerate as acceptors Marinobacter adhaerens HP15 ?
-
 ?
2.4.1.359 additional information the enzyme catalyses the reversible phosphorolysis of 2-O-alpha-D-glucosylglycerol with retention of the anomeric configuration. Homology modelling, docking and mutagenesis are performed to pinpoint particular acceptor site residues (Tyr194, Ala333, Gln336) involved in the binding of glycerol. In the phosphorolytic direction, the enzyme is active on glucosylglycerol but not on sucrose or glucosylglycerate. A variety of other acceptors are surveyed as well, but significant activity can only be observed using 1,2-propanediol. No transglucosylation activity is detected when fructose, fructose 6-phosphate, or glycerate are supplied as acceptor Marinobacter adhaerens HP15 ?
-
-

Subunits

EC Number Subunits Comment Organism
2.4.1.359 ? x * 55900, about, sequence calculation Marinobacter adhaerens

Synonyms

EC Number Synonyms Comment Organism
2.4.1.359 2-O-alpha-D-glucopyranosylglycerol:phosphate alpha-D-glucosyltransferase
-
 Marinobacter adhaerens
2.4.1.359 2-O-alpha-D-glucosylglycerol phosphorylase (retaining)
-
 Marinobacter adhaerens
2.4.1.359 GGoP
-
 Marinobacter adhaerens
2.4.1.359 glucosylglycerol phosphorylase
-
 Marinobacter adhaerens
2.4.1.359 GtfA
-
 Marinobacter adhaerens
2.4.1.359 MaGGoP
-
 Marinobacter adhaerens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.359 37
-
-
 Marinobacter adhaerens

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.1.359 25 50 over 60% of maximal activity within this range Marinobacter adhaerens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.359 45
-
 10 min, no loss of activity Marinobacter adhaerens
2.4.1.359 50
-
 10 min, 46% residual activity Marinobacter adhaerens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.359 3 6 alpha-D-glucose 1-phosphate pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 3 6 alpha-D-glucose 1-phosphate recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 14
-
 phosphate pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 14
-
 2-O-(alpha-D-glucopyranosyl)glycerol pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 14
-
 2-O-(alpha-D-glucopyranosyl)glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 39
-
 glycerol pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 39
-
 glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.359 6.5
-
-
 Marinobacter adhaerens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.359 4.5 7.5 over 20% of maximal activity within this range, profile overview Marinobacter adhaerens

General Information

EC Number General Information Comment Organism
2.4.1.359 evolution the enzyme belongs to the glycoside hydrolase GH13_18 family, simplified phylogenetic tree Marinobacter adhaerens
2.4.1.359 metabolism the enzyme specificity provides additional insights into bacterial metabolic routes, describing a phosphorolytic route for glucosylglycerol in a glucosylglycerol-producing organism Marinobacter adhaerens
2.4.1.359 additional information homology structure modeling using the crystal structure from the Bifidobacterium adolescentis sucrose phosphorylase (PDB IDs 1R7A, 2GDU, 2GDV) as template, docking study and enzyme-substrate interactions analysis, overview. Tyr194 and Gln336 each seemingly form a hydrogen bond with a hydroxyl group of glycerol, while Ala333 is situated at a suitable location for hydrophobic interaction with atom C1' Marinobacter adhaerens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.359 1.4
-
 2-O-(alpha-D-glucopyranosyl)glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 1.4
-
 phosphate pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 3.59
-
 2-O-(alpha-D-glucopyranosyl)glycerol pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 3.6
-
 2-O-(alpha-D-glucopyranosyl)glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 5.57
-
 glycerol pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 5.6
-
 glycerol recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 14.4
-
 alpha-D-glucose 1-phosphate recombinant enzyme, pH 6.5, 37°C Marinobacter adhaerens
2.4.1.359 14.4
-
 alpha-D-glucose 1-phosphate pH 6.5, 37°C Marinobacter adhaerens