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Information on EC 3.2.1.196 - limit dextrin alpha-1,6-maltotetraose-hydrolase for references in articles please use BRENDA:EC3.2.1.196
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EC Tree
IUBMB Comments This bacterial enzyme catalyses a reaction similar to EC 3.2.1.33, amylo-alpha-1,6-glucosidase (one of the activities of the eukaryotic glycogen debranching enzyme). However, while EC 3.2.1.33 removes single glucose residues linked by 1,6-alpha-linkage, and thus requires the additional activity of 4-alpha-glucanotransferase (EC 2.4.1.25) to act on limit dextrins formed by glycogen phosphorylase (EC 2.4.1.1), this enzyme removes maltotriose and maltotetraose chains that are attached by 1,6-alpha-linkage to the limit dextrin main chain, generating a debranched limit dextrin without a need for another enzyme.
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
Hydrolysis of (1->6)-alpha-D-glucosidic linkages to branches with degrees of polymerization of three or four glucose residues in limit dextrin.
Synonyms
GDE, GlgX, GlgX protein, glycogen debranching enzyme, isoamylase-type debranching enzyme,
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glycogen debranching enzyme
isoamylase-type debranching enzyme
GlgX
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gene name
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GlgX protein
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glycogen debranching enzyme
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ambiguous
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glycogen debranching enzyme
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isoamylase-type debranching enzyme
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isoamylase-type debranching enzyme
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isoamylase-type debranching enzyme
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isoamylase-type debranching enzyme
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Hydrolysis of (1->6)-alpha-D-glucosidic linkages to branches with degrees of polymerization of three or four glucose residues in limit dextrin.
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glycogen phosphorylase-limit dextrin maltotetraose-hydrolase
This bacterial enzyme catalyses a reaction similar to EC 3.2.1.33, amylo-alpha-1,6-glucosidase (one of the activities of the eukaryotic glycogen debranching enzyme). However, while EC 3.2.1.33 removes single glucose residues linked by 1,6-alpha-linkage, and thus requires the additional activity of 4-alpha-glucanotransferase (EC 2.4.1.25) to act on limit dextrins formed by glycogen phosphorylase (EC 2.4.1.1), this enzyme removes maltotriose and maltotetraose chains that are attached by 1,6-alpha-linkage to the limit dextrin main chain, generating a debranched limit dextrin without a need for another enzyme.
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amylopectin + H2O
?
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?
amylopectin + H2O
maltose
beta-cyclodextrin-alpha-1,6-linked maltopentaose + H2O
beta-cyclodextrin + maltopentaose
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?
beta-cyclodextrin-alpha-1,6-linked maltotetraose + H2O
beta-cyclodextrin + maltotetraose
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?
beta-cyclodextrin-alpha-1,6-linked maltotriose + H2O
beta-cyclodextrin + maltotriose
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?
G3-beta-cyclodextrin + H2O
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?
G4-beta-cyclodextrin + H2O
?
glycogen + H2O
?
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?
glycogen + H2O
maltose + ?
phosphorylase-limit dextrin
maltodextrin + ?
phosphorylase-limit dextrin + H2O
?
the enzyme hydrolyzes alpha-1,6-glycosidic linkages of phosphorylase-limit dextrin containing only three or four glucose subunits
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raw starch + H2O
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soluble starch + H2O
maltose + ?
additional information
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amylopectin + H2O
maltose
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?
amylopectin + H2O
maltose
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?
G4-beta-cyclodextrin + H2O
?
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?
G4-beta-cyclodextrin + H2O
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GlgX shows substrate specificity for G4 phosphorylase-limit dextrin
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glycogen + H2O
maltose + ?
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?
glycogen + H2O
maltose + ?
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?
phosphorylase-limit dextrin
maltodextrin + ?
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?
phosphorylase-limit dextrin
maltodextrin + ?
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?
soluble starch + H2O
maltose + ?
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?
soluble starch + H2O
maltose + ?
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?
additional information
?
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no activity towards G2-beta-cyclodextrin and G5-beta-cyclodextrin
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additional information
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no activity towards G2-beta-cyclodextrin and G5-beta-cyclodextrin
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additional information
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no activity with G2- and G5-beta-cyclodextrin
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additional information
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no activity with G2- and G5-beta-cyclodextrin
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additional information
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the isoamylase-type debranching enzyme shows high specificity for hydrolysis of chains consisting of three or four glucose residues
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additional information
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the isoamylase-type debranching enzyme shows high specificity for hydrolysis of chains consisting of three or four glucose residues
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phosphorylase-limit dextrin
maltodextrin + ?
phosphorylase-limit dextrin + H2O
?
P15067
the enzyme hydrolyzes alpha-1,6-glycosidic linkages of phosphorylase-limit dextrin containing only three or four glucose subunits
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phosphorylase-limit dextrin
maltodextrin + ?
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?
phosphorylase-limit dextrin
maltodextrin + ?
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?
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0.15
beta-cyclodextrin-alpha-1,6-linked maltotetraose
pH 7, 37°C
1.5
beta-cyclodextrin-alpha-1,6-linked maltotriose
pH 7, 37°C
1.51
G3-beta-cyclodextrin
in 50 mM sodium phosphate buffer (pH7.0) at 37°C
0.15
G4-beta-cyclodextrin
in 50 mM sodium phosphate buffer (pH7.0) at 37°C
additional information
amylopectin
additional information
amylopectin
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KM value 0.056 mg/ml, pH 7.0, 25°C
additional information
glycogen
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KM value 0.062 mg/ml, pH 7.0, 25°C
additional information
starch
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KM value 0.065 mg/ml, pH 7.0, 25°C
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41.7
beta-cyclodextrin-alpha-1,6-linked maltotetraose
pH 7, 37°C
38
beta-cyclodextrin-alpha-1,6-linked maltotriose
pH 7, 37°C
38
G3-beta-cyclodextrin
in 50 mM sodium phosphate buffer (pH7.0) at 37°C
41.7
G4-beta-cyclodextrin
in 50 mM sodium phosphate buffer (pH7.0) at 37°C
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brenda
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brenda
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brenda
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brenda
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brenda
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brenda
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brenda
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brenda
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UniProt
brenda
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brenda
member of glycosyl hydrolase 13 family
UniProt
brenda
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malfunction
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disruption of the glgX gene results in the enhanced fluctuation of glycogen content in the cells during light-dark cycles of the culture
malfunction
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inactivation of glgX leads to slower growth and to a higher intracellular glycogen pool throughout growth, when compared to those in the parental strain
malfunction
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inactivation of glgX leads to slower growth and to a higher intracellular glycogen pool throughout growth, when compared to those in the parental strain
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malfunction
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disruption of the glgX gene results in the enhanced fluctuation of glycogen content in the cells during light-dark cycles of the culture
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metabolism
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the enzyme is predominantly involved in glycogen catabolism by selectively debranching the polysaccharide outer chains that were previously recessed by glycogen phosphorylase
metabolism
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the enzyme is predominantly involved in glycogen catabolism by selectively debranching the polysaccharide outer chains that were previously recessed by glycogen phosphorylase
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physiological function
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the enzyme is required for glycogen degradation and for fast adaptation to hyperosmotic stress
physiological function
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the enzyme is required for glycogen degradation and for fast adaptation to hyperosmotic stress
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76000
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x * 76000, His6-tagged enzyme, SDS-PAGE
78956
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x * 78956, calculated from amino acid sequence
110000
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4 * 110000, His6-tagged enzyme, SDS-PAGE
417000
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native enzyme, gel filtration
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monomer
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1 * 83500 SDS-PAGE
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x * 76000, His6-tagged enzyme, SDS-PAGE
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x * 76000, His6-tagged enzyme, SDS-PAGE
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x * 78956, calculated from amino acid sequence
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x * 78956, calculated from amino acid sequence
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homotetramer
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4 * 110000, His6-tagged enzyme, SDS-PAGE
homotetramer
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4 * 110000, His6-tagged enzyme, SDS-PAGE
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hanging drop vapor diffusion method, using 0.2 M sodium citrate, 47% (v/v) MPD, 4% (v/v) PEG 3350, and HEPES (pH 8.0) buffer; hanging drop vapor diffusion method, using 0.2 M sodium citrate, 47% (w/v) 2-methyl-2,4-pentanediol, 4% (w/v) PEG 3350, and HEPES (pH 8.0) buffer
to 2.25 A resolution. Structure reveals a monomer consisting of three major domains with high structural similarity to the subunit of TreX, the oligomeric bifunctional glycogen debranching enzyme from Sulfolobus. In the overlapping substrate binding groove, conserved residues Leu270, Asp271, and Pro208 block the cleft, yielding a shorter narrow GlgX cleft compared to that of TreX. Residues 207-213 form a unique helical conformation that is observed in both GlgX and TreX, possibly distinguishing GDEs from isoamylases and pullulanases
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45
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1 h, 59% residual activity
95
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the His6-tagged enzyme is inactivated by incubation at 95°C for 10 min
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HisTrap column chromatography and Superdex 200 gel filtration
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Ni-NTA agarose column chromatography
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Ni-NTA column chromatography and Superdex 200 gel filtration; Ni-NTA column chromatography and Superdex 200 gel filtration
purification from culture filtrate
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expressed in Escherichia coli BL21AI cells
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expressed in Escherichia coli ER2566 cells
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expressed in Escherichia coli MC1061 cells; expressed in Escherichia coli MC1061 cells
expressed in the glgX mutant of Synechocystis elongatus PCC 7942
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maximum gene transcription is observed after 5 min of incubation with seed exudates (50.29fold) whereas in the presence of naringenin a 2.88fold increase is observed after 8 h of incubation
maximum gene transcription is observed after 5 min of incubation with seed exudates (50.29fold) whereas in the presence of naringenin a 2.88fold increase is observed after 8 h of incubation
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maximum gene transcription is observed after 5 min of incubation with seed exudates (50.29fold) whereas in the presence of naringenin a 2.88fold increase is observed after 8 h of incubation
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Song, H.N.; Jung, T.Y.; Park, J.T.; Park, B.C.; Myung, P.K.; Boos, W.; Woo, E.J.; Park, K.H.
Structural rationale for the short branched substrate specificity of the glycogen debranching enzyme GlgX
Proteins
78
1847-1855
2010
Escherichia coli, Escherichia coli (P15067)
brenda
Song, H.; Jung, T.; Park, J.; Park, B.; Myung, P.; Boos, W.; Woo, E.; Park, K.
Structural ratio-rials for the short branched substrato specificity of the glycogen debranching enzyme GlgX
Proteins Struct. Funct. Bioinform.
78
1847-1855
2010
Escherichia coli (P15067)
brenda
Suzuki, E.; Umeda, K.; Nihei, S.; Moriya, K.; Ohkawa, H.; Fujiwara, S.; Tsuzuki, M.; Nakamura, Y.
Role of the GlgX protein in glycogen metabolism of the cyanobacterium, Synechococcus elongatus PCC 7942
Biochim. Biophys. Acta
1770
763-773
2007
Synechococcus elongatus, Synechococcus elongatus PCC 6803
brenda
Oliveira, L.R.; Marcelino, F.C.; Barcellos, F.G.; Rodrigues, E.P.; Megias, M.; Hungria, M.
The nodC, nodG, and glgX genes of Rhizobium tropici strain PRF 81
Funct. Integr. Genomics
10
425-431
2010
Rhizobium tropici, Rhizobium tropici PRF 81
brenda
Dauvillee, D.; Kinderf, I.S.; Li, Z.; Kosar-Hashemi, B.; Samuel, M.S.; Rampling, L.; Ball, S.; Morell, M.K.
Role of the Escherichia coli glgX gene in glycogen metabolism
J. Bacteriol.
187
1465-1473
2005
Escherichia coli, Escherichia coli BW25113
brenda
Seibold, G.M.; Eikmanns, B.J.
The glgX gene product of Corynebacterium glutamicum is required for glycogen degradation and for fast adaptation to hyperosmotic stress
Microbiology
153
2212-2220
2007
Corynebacterium glutamicum, Corynebacterium glutamicum ATCC 13032
brenda
Shetty, P.
Glucoamylase from the predacious fungus Arthrobotrys conoides a cationic enzyme with high debranching activity and raw starch digestibility
Appl. Biochem. Microbiol.
52
176-182
2016
Arthrobotrys conoides
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brenda
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3.2.1.196 limit dextrin alpha-1,6-maltotetraose-hydrolase
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