Any feedback?
Information on EC 3.2.1.142 - limit dextrinase
for references in articles please use BRENDA:EC3.2.1.142Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
3.2.1.142 limit dextrinaseIUBMB Comments
Plant enzymes with little or no action on glycogen. Action on amylopectin is incomplete, but action on alpha-limit dextrins is complete. Maltose is the smallest sugar it can release from an alpha-(1->6)-linkage.
Specify your search results
Word Map
- 3.2.1.142
- barley
- malt
- pullulan
- amylopectin
- isoamylase
-
diastatic
-
3.2.1.41
- pullulanases
- analysis
-
starch-branching
-
beta-limit
- food industry
- beta-amylase
- synthesis
- brewing
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
Synonyms
limit dextrinase, starch-debranching enzyme, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
pullulanase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hyrolysis of O-glycosyl bonds
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
dextrin 6-alpha-glucanohydrolase
Plant enzymes with little or no action on glycogen. Action on amylopectin is incomplete, but action on alpha-limit dextrins is complete. Maltose is the smallest sugar it can release from an alpha-(1->6)-linkage.
CAS REGISTRY NUMBER
COMMENTARY
9032-15-9
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
limit dextrin + H2O
?
the enzyme hydrolyses alpha-1,6-glucosidic linkages in limit dextrins derived from amylopectin
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
limit dextrin + H2O
?
the enzyme hydrolyses alpha-1,6-glucosidic linkages in limit dextrins derived from amylopectin
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
amylopectin
Km value of wild-type 6.9 mg/ml, mutant M440G 4.4 mg/ml, respectively, 37°C, pH not specified in the publication
additional information
amylopectin
-
Km value of wild-type 6.9 mg/ml, mutant M440G 4.4 mg/ml, respectively, 37°C, pH not specified in the publication
additional information
additional information
classical Michaelis-Menten kinetics results in Km of 0.081 mg/ml, fitting the uncompetitive substrate inhibition Michaelis-Menten equation results in Km 0.16 mg/ml, pH 5.5, 40°C
-
additional information
additional information
-
classical Michaelis-Menten kinetics results in Km of 0.081 mg/ml, fitting the uncompetitive substrate inhibition Michaelis-Menten equation results in Km 0.16 mg/ml, pH 5.5, 40°C
-
additional information
pullulan
Km value of wild-type 0.16 mg/ml, mutant M440G 0.15 mg/ml, respectively, 37°C, pH not specified in the publication
additional information
pullulan
-
Km value of wild-type 0.16 mg/ml, mutant M440G 0.15 mg/ml, respectively, 37°C, pH not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.9
amylopectin
mutant M440G, 37°C, pH not specified in the publication
78
pullulan
fitting the uncompetitive substrate inhibition Michaelis-Menten equation, pH 5.5, 40°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
amylopectin
kcat/Km value of wild-type 2.3 ml/mg/s, mutant M440G 0.9 ml/mg/s, respectively, 37°C, pH not specified in the publication
additional information
amylopectin
-
kcat/Km value of wild-type 2.3 ml/mg/s, mutant M440G 0.9 ml/mg/s, respectively, 37°C, pH not specified in the publication
additional information
additional information
classical Michaelis-Menten kinetics results in kcat/Km 753 ml/mg s, fitting the uncompetitive substrate inhibition Michaelis-Menten equation results in kcat/Km 488 ml/mg s, pH 5.5, 40°C
-
additional information
additional information
-
classical Michaelis-Menten kinetics results in kcat/Km 753 ml/mg s, fitting the uncompetitive substrate inhibition Michaelis-Menten equation results in kcat/Km 488 ml/mg s, pH 5.5, 40°C
-
additional information
pullulan
kcat/Km value of wild-type 488 ml/mg/s, mutant M440G 480 ml/mg/s, respectively, 37°C, pH not specified in the publication
additional information
pullulan
-
kcat/Km value of wild-type 488 ml/mg/s, mutant M440G 480 ml/mg/s, respectively, 37°C, pH not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the debranching enzyme belongs to the glycoside hydrolase family 13 subfamily 13, GH13_13
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). O48541_HORVV
904
0
99124
TrEMBL
other Location (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
binding of alpha-, beta-, and gamma-cyclodextrin binding to limit dextrinase with Kd of 27.2, 0.70, and 34.7 microM, respectively
additional information
-
binding of alpha-, beta-, and gamma-cyclodextrin binding to limit dextrinase with Kd of 27.2, 0.70, and 34.7 microM, respectively
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with its endogenous inhibitor LDI, at 2.7 A resolution. A hydrophobic cluster flanked by ionic interactions in the protein-protein interface is vital for the picomolar affinity of LDI to the enzyme
purified free enzyme with a glycerol molecule in the active site or purified enzyme in complex with competitive inhibitors alpha- and beta-cyclodextrin, hanging drop vapour diffusion method, streak-seeding, mixing of 10 mg/mlprotein in 50 mM MES buffer pH 6.6, 250 mM NaCl, 0.5 mM CaCl2, 0.67mM maltotriose, with a reservoir solution consisting of 30% w/v PEG 3350, 5% glycerol, 0.3 M NaI, cysteine is added to the crystallization drops to a final concentration of 5-7 mM, one week, 20°C, X-ray diffraction structure determination and analysis at 1.9-2.5 A resolution, molecular replacement and modelling
structures of barley limit dextrinase and active-site mutants in complex with natural substrates, products and substrate analogues. Avoidance of alpha-1,4-hydrolytic activity and strong preference for alpha-1,6-hydrolytic activity seems to be based on the strong interaction to Trp512 and Phe553 on each flank of the catalytic cleft, which will keep non-branched polysaccharides out of the reach of the catalytic nucleophile and acid-base catalyst
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A885S
single nucleotide polymorphism associated with thermostability
D730R
modest change in the association rate constant with endogenous inhibitor LDI. Residue D730 binds in a positively charged pocket on the surface of LDI via a hydrogen bond and a salt bridge to residue Arg34 and a hydrogen bond to Arg84 of LDI. D730 adopts different rotamers in the free enzyme and enzyme bound to LDI
D730W
modest change in the association rate constant with endogenous inhibitor LDI. Residue D730 binds in a positively charged pocket on the surface of LDI via a hydrogen bond and a salt bridge to residue Arg34 and a hydrogen bond to Arg84 of LDI. D730 adopts different rotamers in the free enzyme and enzyme bound to LDI
T233A
single nucleotide polymorphism associated with thermostability
additional information
additional information
investigation on genetic diversity of limit dextrinase using single strand conformation polymorphism based on the amino acid substitutions. Only limited genetic variation is detected in the current malting barley varieties, although wide variation is observed in the wider barley germplasm
additional information
investigation on genetic diversity of limit dextrinase using single strand conformation polymorphism based on the amino acid substitutions. Only limited genetic variation is detected in the current malting barley varieties, although wide variation is observed in the wider barley germplasm
additional information
investigation on genetic diversity of limit dextrinase using single strand conformation polymorphism based on the amino acid substitutions. Only limited genetic variation is detected in the current malting barley varieties, although wide variation is observed in the wider barley germplasm
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
the level of limit dextrinase thermostability is inversely correlated with total limit dextrinase activity
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
methods for a rapid and cost efficient assay of both beta-amylase and limit dextrinase thermostability
food industry
the prediction of malt fermentability is achieved by both forward step-wise multi-linear regression and the partial least squares multivariate model development methods. Both methods produce similar identifications of the parameters predicting wort fermentability at similar levels of predictive power. Both models are substantially better at predicting fermentability than the traditional use of diastatic power on its own. Limit dextrinase thermostability is not a substantial predictor of fermentability, presumably due to its negative correlation with total limit dextrinase activity. The application of these insights in the malting and brewing industries is expected to result in substantial improvements in brewing consistency and enable more specific quality targets for barley breeders progeny selection cut-off limits to be more precisely defined
synthesis
expression of the limit dextrinase encoding gene fragment without signal peptide, with the Saccharomyces cerevisiae alpha-factor secretion signal under control of the alcohol oxidase 1 promoter, in Pichia pastoris leads to highly active barley limit dextrinase secreted during high cell-density fermentation. Optimization of a fedbatch fermentation procedure enables efficient production in a 5-l bioreactor, yielding 34 mg homogenous enzyme with 84% recovery
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Evans, D.; Dambergs, R.; Ratkowsky, D.; Li, C.; Harasymow, S.; Roumeliotis, S.; Eglinton, J.
Refining the prediction of potential malt fermentability by including an assessment of limit dextrinase thermostability and additional measures of malt modification, using two different methods for multivariate model development
J. Inst. Brew.
116
86-96
2010
Hordeum vulgare (O48541)
-
Yang, X.; Westcott, S.; Gong, X.; Evans, E.; Zhang, X.; Lance, R.; Li, C.
Amino acid substitutions of the limit dextrinase gene in barley are associated with enzyme thermostability
Mol. Breed.
23
61-74
2009
Hordeum vulgare (O48541), Hordeum vulgare (Q9FYY0), Hordeum vulgare (Q9S7S8)
-
Vester-Christensen, M.B.; Hachem, M.A.; Naested, H.; Svensson, B.
Secretory expression of functional barley limit dextrinase by Pichia pastoris using high cell-density fermentation
Protein Expr. Purif.
69
112-119
2010
Hordeum vulgare (O48541), Hordeum vulgare
Moeller, M.S.; Abou Hachem, M.; Svensson, B.; Henriksen, A.
Structure of the starch-debranching enzyme barley limit dextrinase reveals homology of the N-terminal domain to CBM21
Acta Crystallogr. Sect. F
68
1008-1012
2012
Hordeum vulgare (O48541), Hordeum vulgare
Moller, M.S.; Vester-Christensen, M.B.; Jensen, J.M.; Hachem, M.A.; Henriksen, A.; Svensson, B.
Crystal structure of barley limit dextrinase-limit dextrinase inhibitor (LD-LDI) complex reveals insights into mechanism and diversity of cereal type inhibitors
J. Biol. Chem.
290
12614-12629
2015
Hordeum vulgare (O48541), Hordeum vulgare
Moller, M.S.; Windahl, M.S.; Sim, L.; Bojstrup, M.; Abou Hachem, M.; Hindsgaul, O.; Palcic, M.; Svensson, B.; Henriksen, A.
Oligosaccharide and substrate binding in the starch debranching enzyme barley limit dextrinase
J. Mol. Biol.
427
1263-1277
2015
Hordeum vulgare (O48541), Hordeum vulgare
EXTERNAL LINKS (specific for EC-Number 3.2.1.142)
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
