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Information on EC 2.4.1.4 - amylosucrase and Organism(s) Neisseria polysaccharea and UniProt Accession Q9ZEU2
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2.4.1.4 amylosucraseIUBMB Comments
The glucansucrases transfer a D-glucosyl residue from sucrose to a glucan chain. They are classified based on the linkage by which they attach the transferred residue. In some cases, in which the enzyme forms more than one linkage type, classification relies on the relative proportion of the linkages that are generated. This enzyme extends the glucan chain by an alpha(1->4) linkage.
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Word Map
- 2.4.1.4
- neisseria
- polysaccharea
-
deinococcus
- geothermalis
- synthesis
- food industry
- biotechnology
-
amylose-like
-
transglucosylation
-
waxy
- drug development
- turanose
- asases
- transglucosidase
- amylopectin
-
c-myc-binding
- maltooligosaccharides
-
glycoside-hydrolase
- trehalulose
- industry
The taxonomic range for the selected organisms is: Neisseria polysaccharea
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Reaction Schemes
Synonyms
amylosucrase, amy-1, drpas, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
glucosyltransferase, sucrose-1,4-alpha-glucan
-
-
-
-
sucrose-glucan glucosyltransferase
-
-
-
-
additional information
additional information
the enzyme is a glucansucrase from glycoside hydrolase, GH, family 13
additional information
the enzyme is a member of family 13 of the glycoside hydrolases
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sucrose + [(1->4)-alpha-D-glucosyl]n = D-fructose + [(1->4)-alpha-D-glucosyl]n+1
in Neisseria polysaccharea NpAS key residues force the fructosyl moiety to bind in an open state with the O3' ideally positioned to explain the preferential formation of turanose by NpAS. Residues Glu328 and Asp286 of NpAS are the general acid/base and the nucleophile, respectively, involved in the formation of the beta-glucosyl intermediate occurring in the alpha-retaining mechanism
sucrose + [(1->4)-alpha-D-glucosyl]n = D-fructose + [(1->4)-alpha-D-glucosyl]n+1
alpha-retaining mechanism via a double-displacement similar to that described for alpha-amylases
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexosyl group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
SYSTEMATIC NAME
IUBMB Comments
sucrose:(1->4)-alpha-D-glucan 4-alpha-D-glucosyltransferase
The glucansucrases transfer a D-glucosyl residue from sucrose to a glucan chain. They are classified based on the linkage by which they attach the transferred residue. In some cases, in which the enzyme forms more than one linkage type, classification relies on the relative proportion of the linkages that are generated. This enzyme extends the glucan chain by an alpha(1->4) linkage.
CAS REGISTRY NUMBER
COMMENTARY
9032-11-5
-
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
n sucrose
[(1->4)-alpha-D-glucosyl]n + n fructose
80.1% polymerization products, additionally ASase catalyzes reactions with 14.5% isomerization products and 5.4% hydrolysis products
-
-
?
sucrose
glucose + maltose + maltotriose + turanose + insoluble polymer
enzyme catalyzes both sucrose hydrolysis and oligosaccharide and polymer synthesis in the absence of an activator polymer
with 10 mM sucrose as the sole substrate, 30% glucose, 29% maltose, 18% maltotriose, 11% turanose and 12% insoluble polymer respectively
?
sucrose
maltose + maltotriose + turanose + erlose
compared to the wild-type enzyme, and in agreement with their loss of polymerase activity, all three mutant enzymes incorporate higher amounts of glucosyl units in maltose (27.3% (mutant enzyme R226L/I228V/F229A/A289I/F290Y/E300I/V331T); 24.8% (mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D), 15.3% (mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R), versus 5.8% for wild-type enzyme) and in maltotriose (20.5% (mutant R226L/I228V/F229A/A289I/F290Y/E300I/V331T); 18.6% (mutant R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D), 23% (mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R), versus 2.9% for wild-type enzyme). Compared to the others, the mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R is more specialized in turanose production, incorporating nearly 46% of the glucosyl residues in turanose, versus only 19% for the wild-type enzyme. With mutants enzyme R226L/I228V/F229A/A289I/F290Y/E300I/V331T and mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D, 20% and 23% glucosyl units are incorporated into erlose (alpha-D-glucopyranosyl-(1->4)-alpha-D-glucopyranosyl-(1->2)-beta-D-fructose), respectively. Much lower values are observed with mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R (only 1.4%) and none for the wild-type enzyme. Panose (alpha-D-glucopyranosyl-(1->6)-alpha-D-glucopyranosyl-(1->4)-alpha-D-glucose) is mainly produced by mutants R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D and R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R, 13.9% and 8.5% of the glucosyl units incorporated into this trisaccharide, respectively. In comparison, the value goes down to 1.6% with mutant enzyme R226L/I228V/F229A/A289I/F290Y/E300I/V331T and it is not produced by the wild-type enzyme
-
-
?
sucrose + (+)-taxifolin
D-fructose + (+)-taxifolin-4'-O-alpha-D-glucopyranoside
-
-
-
?
sucrose + (-)-epicatechin
D-fructose + (-)-epicatechin-3'-O-alpha-D-glucopyranoside
-
-
-
?
sucrose + (-)-epicatechin-3'-O-alpha-D-glucopyranoside
D-fructose + (-)-epicatechin-3'-O-alpha-D-maltoside
-
-
-
?
sucrose + aesculetin
D-fructose + aesculetin 7-alpha-D-glucopyranoside
-
-
-
?
sucrose + aesculetin 7-alpha-D-glucopyranoside
D-fructose + aesculetin 7-alpha-D-maltoside
-
-
-
?
sucrose + aesculetin 7-alpha-D-maltoside
D-fructose + aesculetin 7-alpha-D-maltotrioside
-
-
-
?
sucrose + aesculin
D-fructose + aesculin 4-alpha-glucoside
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-
-
?
sucrose + aesculin 4-alpha-glucoside
D-fructose + aesculin 4-alpha-maltoside
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-
-
?
sucrose + alpha-D-glucopyranosyl-(1->4)-salicin
D-fructose + alpha-D-glucopyranosyl-(1->4)-alpha-D-glucopyranosyl-(1->4)-salicin
-
-
-
?
sucrose + amylopectin
?
waxy corn starch selcted as acceptor. The chain length distribution of the elongated waxy corn starchs indicates that all of the branch chains of waxy corn starch are greatly elongated by amylosucrase before occurrence of starch precipitation. Afterwards, however, amylosucrase merely elongates the branch chains whose non-reducing ends are exposed on the surface of the precipitate
-
-
?
sucrose + epicatechin-3'-O-alpha-D-maltoside
D-fructose + (-)-epicatechin-3'-O-alpha-D-maltotrioside
-
-
-
?
sucrose + luteolin
D-fructose + luteolin-4'-O-alpha-D-glucopyranoside
-
-
-
?
sucrose + phloretin
D-fructose + phloretin glucoside 1 + phloretin glucoside 2 + phloretin glucoside 3
the enzyme catalyzes the stereospecific glucosylation of phloretin at the 4'-position
-
-
?
sucrose + phloretin
D-fructose + phloretin glucoside A1 + phloretin glucoside A2 + phloretin glucoside A3
the enzyme is a non-Leloir glycosyltransferase that catalyzes the stereospecific glucosylation of phloretin at the 4'-position. Phloretin and its glucosylation derivatives are cytotoic, overview
three major phloretin-dependent sugar-positive products are observed containing one to three Glc residues (Phlo-A1, -A2, -A3), identification by TLC and NMR spectrometry. In all three metabolites the first Glc, GlcA, is linked to the aglycone at C4'
-
?
sucrose + phloretin
D-fructose + phloretin-4'-O-alpha-D-glucopyranoside
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-
-
?
sucrose + phloretin-4'-O-alpha-D-glucopyranoside
D-fructose + phloretin-4'-O-alpha-D-maltoside
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-
-
?
sucrose + phloretin-4'-O-alpha-D-maltoside
D-fructose + phloretin-4'-O-alpha-D-maltotrioside
-
-
-
?
sucrose + salicin
D-fructose + alpha-D-glucopyranosyl-(1,4)-salicin + alpha-D-glucopyranosyl-(1,4)-alpha-D-glucopyranosyl-(1,4)-salicin
synthesis of salicin glycosides with sucrose serving as the glucopyranosyl donor and salicin as the acceptor molecule
i.e. glucosyl salicin and maltosyl salicin, identification by NMR and TLC analysis
-
?
sucrose + salicin
D-fructose + alpha-D-glucopyranosyl-(1->4)-salicin
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-
-
?
sucrose + vanillin
D-fructose + vanillin 4-alpha-D-glucopyranoside
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-
-
?
sucrose + zingerone
D-fructose + zingerone 4-alpha-D-glucopyranoside
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-
-
?
sucrose + amylose
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
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-
-
-
?
sucrose + maltopentaose
D-fructose + maltohexaose + maltoheptaose
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-
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
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linear alpha-(1,4)-glucans
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
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-
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
the enzyme catalyzes the synthesis of alpha-1,4 glucans from sucrose. The product profile is quite polydisperse, ranging from soluble chains called maltooligosaccharides to high-molecular weight insoluble amylose
-
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A only produces soluble oligosaccharides as no insoluble high molecular weight amylose is observed
-
-
?
sucrose + (1,4-alpha-D-glucosyl)n
D-fructose + (1,4-alpha-D-glucosyl)n+1
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-
recombinant enzyme linearly elongates some branched chains of glycogen to an average degree of polymerization of 75
?
sucrose + (1,4-alpha-D-glucosyl)n
D-fructose + (1,4-alpha-D-glucosyl)n+1
-
-
recombinant enzyme produces glucopolysaccharide mainly composed of alpha-(1-4) glucosidic linkages and a very low degree, i.e. less than 5%, of alpha-(1-6) branched linkages
?
sucrose + (1,4-alpha-D-glucosyl)n
D-fructose + (1,4-alpha-D-glucosyl)n+1
-
amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size are produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation
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-
?
sucrose + (1,4-alpha-D-glucosyl)n
D-fructose + (1,4-alpha-D-glucosyl)n+1
-
glycogen is the best D-glucosyl unit acceptor. Semiprocessive glycogen elongation mechanism can be proposed on the basis of modeling data
-
-
?
additional information
?
-
the enzyme catalyzes the synthesis of a water-insoluble amylose-like polymer from sucrose, a readily available and low-cost agroresource
-
-
?
additional information
?
-
the enzyme catalyze the synthesis of an alpha-(1,4)-linked glucan polymer from sucrose instead of an expensive activated sugar, such as ADP- or UDP-glucose
-
-
?
additional information
?
-
-
the enzyme catalyze the synthesis of an alpha-(1,4)-linked glucan polymer from sucrose instead of an expensive activated sugar, such as ADP- or UDP-glucose
-
-
?
additional information
?
-
product patterns formed by wild-type enzyme and selected genetic variants in the presence of sucrose as the sole substrate, overview
-
-
?
additional information
?
-
synthesis of sucrose isomers turanose and trehalulose from sucrose in the presence of fructose by NpAS, turanose binding site structure, overview
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-
?
additional information
?
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synthesis of sucrose isomers turanose and trehalulose from sucrose in the presence of fructose by NpAS, turanose binding site structure, overview
-
-
?
additional information
?
-
amylosucrase (AS), a glucosyltransferase from Neiserria polysaccharea, produces an insoluble alpha-1,4-linked glucan polymer by consuming sucrose and releasing fructose. This reaction does not require a-D-glucosyl-nucleotide-diphosphate like ADP- or UDP-glucose, but rather uses the energy generated by splitting sucrose in order to synthesise the glucan polymer
-
-
?
additional information
?
-
amylosucrase synthesizes alpha-1,4-glucans using sucrose as a sole substrate
-
-
?
additional information
?
-
-
amylosucrase synthesizes alpha-1,4-glucans using sucrose as a sole substrate
-
-
?
additional information
?
-
the amylosucrase from Neisseria polysaccharea naturally catalyzes the synthesis of alpha-glucans from the widely available donor sucrose. NpAS is highly specific for its natural substrate and subsite -1 (according to GH nomenclature) plays a major role in the recognition of the sucrose glucosyl moiety through a highly efficient hydrogen bonding interaction network, subsite 21 is responsible for the high affinity for sucrose
-
-
?
additional information
?
-
-
the amylosucrase from Neisseria polysaccharea naturally catalyzes the synthesis of alpha-glucans from the widely available donor sucrose. NpAS is highly specific for its natural substrate and subsite -1 (according to GH nomenclature) plays a major role in the recognition of the sucrose glucosyl moiety through a highly efficient hydrogen bonding interaction network, subsite 21 is responsible for the high affinity for sucrose
-
-
?
additional information
?
-
analysis of enzyme substrate specificity, from 11 potential donors harboring selective derivatizations that are experimentally evaluated, only 4-nitrophenyl-alpha-D-glucopyranoside is used by the wild-type enzyme, and this underlines the high specificity of the -1 subsite of enzyme NpAS for glucosyl donor substrates. Acceptor substrate promiscuity is explored by screening 20 hydroxylated molecules, including D- and L-monosaccharides as well as polyols. With the exception of one compound, all are successfully glucosylated, and showig the tremendous plasticity of the +1 subsite of NpAS, which is responsible for acceptor recognition. Acceptor substrates are arabinose, galactose, altrose, fucose, xylose, allose, mannose, D-sorbitol, Darabitol, D-mannitol, xylitol, myo-inositol, and maltitol. Analysis of product structures and enzyme enantiopreference by in silico docking analyses. The enzyme is able to discriminate very similar molecules such as enantiomers. Arabinose and altrose are more efficiently glucosylated by NpAS in their L forms, whereas xylose is better recognized in its D form. Glucosylation of mannose, xylose, and galactose are less discriminant, while the enzyme isstrictly enantiospecific toward D-fucose
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-
?
additional information
?
-
crystalline structures of waxy corn starch treated with the enzyme, detailed overview. The crystalline structures in the amylosucrase-modified starch are the result of the formation of intermolecular double helices among amylopectins with elongated external chains. The degree of mutual binding by hydrogen bonds between amylopectins is responsible for the amount of crystalline structure. When these bonds are strong and numerous, the chains associate as crystalline structures, resulting in high SDS and/or RS content. The internal structures of AS-modified starch are not significantly different from the control. This is a plausible explanation for the insignificant change in RS content of the AS-modified starches with the varying reaction times
-
-
?
additional information
?
-
hydrolysis of p-nitrophenyl-alpha-D-glucopyranoside is used for activity measurements. Substrate specificities of recombinant wild-type and mutant enzymes, overview
-
-
?
additional information
?
-
-
hydrolysis of p-nitrophenyl-alpha-D-glucopyranoside is used for activity measurements. Substrate specificities of recombinant wild-type and mutant enzymes, overview
-
-
?
additional information
?
-
-
the recombinant amylosucrase is used to glucosylate glycogen particles in vitro in the presence of sucrose as the glucosyl donor. The morphology and structure of the resulting insoluble products are shown to strongly depend on the initial sucrose/glycogen weight ratio. For the lower ratio (1.14), all glucose molecules produced from sucrose are transferred onto glycogen, yielding a slight elongation of the external chains and their organization into small crystallites at the surface of the glycogen particles. With a high initial sucrose/glycogen ratio (342), the external glycogen chains are extended by amylosucrase, yielding dendritic nanoparticles with a diameter 4-5 times that of the initial particle
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-
?
additional information
?
-
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synthesis of cycloamyloses from sucrose by dual enzyme treatment via combined reaction of amylosucrase and 4-alpha-glucanotransferase from Synechocystis sp., EC 2.4.1.25, overview
-
-
?
additional information
?
-
-
amylosucrase from Neisseria polysaccharea is a transglucosylase that synthesizes an insoluble amylose-like polymer from sole substrate sucrose, product isolation and analysis, overview
-
-
?
additional information
?
-
-
treatment of pre-gelatinized rice and barley starches with amylosucrase from Neisseria polysaccharea for resistant starch production. Analysis of reaction efficiency, resistant starch content, amylopectin branch-chain length distribution, solubility, welling power, pasting viscosity, and thermal transition properties, detailed overview
-
-
?
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
sucrose + phloretin
D-fructose + phloretin glucoside A1 + phloretin glucoside A2 + phloretin glucoside A3
the enzyme is a non-Leloir glycosyltransferase that catalyzes the stereospecific glucosylation of phloretin at the 4'-position. Phloretin and its glucosylation derivatives are cytotoic, overview
three major phloretin-dependent sugar-positive products are observed containing one to three Glc residues (Phlo-A1, -A2, -A3), identification by TLC and NMR spectrometry. In all three metabolites the first Glc, GlcA, is linked to the aglycone at C4'
-
?
sucrose + (1,4-alpha-D-glucosyl)n
D-fructose + (1,4-alpha-D-glucosyl)n+1
-
amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size are produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation
-
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
-
-
linear alpha-(1,4)-glucans
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
-
-
-
?
sucrose + [(1->4)-alpha-D-glucosyl]n
D-fructose + [(1->4)-alpha-D-glucosyl]n+1
the enzyme catalyzes the synthesis of alpha-1,4 glucans from sucrose. The product profile is quite polydisperse, ranging from soluble chains called maltooligosaccharides to high-molecular weight insoluble amylose
-
-
?
additional information
?
-
the enzyme catalyzes the synthesis of a water-insoluble amylose-like polymer from sucrose, a readily available and low-cost agroresource
-
-
?
additional information
?
-
the enzyme catalyze the synthesis of an alpha-(1,4)-linked glucan polymer from sucrose instead of an expensive activated sugar, such as ADP- or UDP-glucose
-
-
?
additional information
?
-
-
the enzyme catalyze the synthesis of an alpha-(1,4)-linked glucan polymer from sucrose instead of an expensive activated sugar, such as ADP- or UDP-glucose
-
-
?
additional information
?
-
amylosucrase (AS), a glucosyltransferase from Neiserria polysaccharea, produces an insoluble alpha-1,4-linked glucan polymer by consuming sucrose and releasing fructose. This reaction does not require a-D-glucosyl-nucleotide-diphosphate like ADP- or UDP-glucose, but rather uses the energy generated by splitting sucrose in order to synthesise the glucan polymer
-
-
?
additional information
?
-
amylosucrase synthesizes alpha-1,4-glucans using sucrose as a sole substrate
-
-
?
additional information
?
-
-
amylosucrase synthesizes alpha-1,4-glucans using sucrose as a sole substrate
-
-
?
additional information
?
-
the amylosucrase from Neisseria polysaccharea naturally catalyzes the synthesis of alpha-glucans from the widely available donor sucrose. NpAS is highly specific for its natural substrate and subsite -1 (according to GH nomenclature) plays a major role in the recognition of the sucrose glucosyl moiety through a highly efficient hydrogen bonding interaction network, subsite 21 is responsible for the high affinity for sucrose
-
-
?
additional information
?
-
-
the amylosucrase from Neisseria polysaccharea naturally catalyzes the synthesis of alpha-glucans from the widely available donor sucrose. NpAS is highly specific for its natural substrate and subsite -1 (according to GH nomenclature) plays a major role in the recognition of the sucrose glucosyl moiety through a highly efficient hydrogen bonding interaction network, subsite 21 is responsible for the high affinity for sucrose
-
-
?
additional information
?
-
-
amylosucrase from Neisseria polysaccharea is a transglucosylase that synthesizes an insoluble amylose-like polymer from sole substrate sucrose, product isolation and analysis, overview
-
-
?
additional information
?
-
-
treatment of pre-gelatinized rice and barley starches with amylosucrase from Neisseria polysaccharea for resistant starch production. Analysis of reaction efficiency, resistant starch content, amylopectin branch-chain length distribution, solubility, welling power, pasting viscosity, and thermal transition properties, detailed overview
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
glycogen
-
initial sucrose consumption rate strongly increases with glycogen concentration, at 106 mM sucrose, addition of 30 g/l glycogen increases the initial rate 98fold
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
78.43
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R
128.3
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A
38.7
sucrose
-
initial sucrose concentration higehr than 20 mM, sucrose hydrolysis
50.2
sucrose
-
initial sucrose concentration higher than 20 mM, sucrose consumption
387
sucrose
-
initial sucrose concentration higher than 20 mM, polymerization
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.61
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A
4.5
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R
0.167
sucrose
-
initial sucrose concentration lower than 20 mM, polymerization
0.333
sucrose
-
initial sucrose concentration lower than 20 mM, sucrose hydrolysis
0.55
sucrose
-
initial sucrose concentration lower than 20 mM, sucrose consumption
0.55
sucrose
-
initial sucrose concentration higher than 20 mM, sucrose hydrolysis
1.28
sucrose
-
initial sucrose concentration higher than 20 mM, sucrose consumption
1.88
sucrose
-
initial sucrose concentration higher than 20 mM, polymerization
3.1
sucrose
-
with maltobiose, pH 7.0, 30°C, recombinant wild-type enzyme
4.8
sucrose
-
with maltotriose, pH 7.0, 30°C, recombinant wild-type enzyme
7.5
sucrose
-
with maltotetraose, pH 7.0, 30°C, recombinant wild-type enzyme
33
sucrose
-
with maltotriose, pH 7.0, 30°C, recombinant mutant R226N
47
sucrose
-
with maltopentaose, pH 7.0, 30°C, recombinant wild-type enzyme
67
sucrose
-
with maltotetraose, pH 7.0, 30°C, recombinant mutant R226N
173
sucrose
-
with maltopentaose, pH 7.0, 30°C, recombinant mutant R226N
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.012
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D
0.013
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A
0.015
sucrose
30°C, pH 7.0, mutant enzyme R226L/I228V/F229A/A289I/F290Y/E300I/V331T
0.058
sucrose
30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5 - 9.5
activity range, wild-type and mutant enzymes, profile overview
7 - 8.5
-
60% of maximal activity at pH 7.0, maximal activity at pH 8.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
35
35
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20 - 50
recombinant enzyme, activity range, temperature profile, overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
physiological function
amylosucrases are sucrose-utilizing alpha-transglucosidases that naturally catalyze the synthesis of alpha-glucans, linked exclusively through alpha-1,4-linkages. Side products and in particular sucrose isomers such as turanose and trehalulose are also produced by these enzymes
evolution
-
the enzyme belongs to the GH13 family. Amylosucrases adopt a deep pocket topology of about 15 A with the catalytic triad located at the bottom
additional information
evolution
the enzyme belongs to the glycohydrolase family 13, GH13
evolution
amylosucrase from Neisseria polysaccharea is a transglucosidase from the GH13 family of glycoside-hydrolases. Natural molecular evolution has modeled a dense hydrogen bond network at subsite 21 responsible for the specific recognition of sucrose and conversely, it has loosened interactions at the subsite 11 creating a highly promiscuous subsite 11. The residues forming these subsites are considered to be likely involved in the activity as well as the overall stability of the enzyme
additional information
the catalytic domain A of NpAS adopts the typical (alpha/alpha)8-barrel of the GH family 13
additional information
-
the catalytic domain A of NpAS adopts the typical (alpha/alpha)8-barrel of the GH family 13
additional information
essential role of enzyme subsite -1 for protein fitness and robustness
additional information
-
essential role of enzyme subsite -1 for protein fitness and robustness
additional information
-
active site topology and structure coparisons, overview
additional information
-
Arg226, was proposed from molecular modeling studies to play an important role in the formation of the active site topology and in the accessibility of ligands to the catalytic site
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). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
72466
x * 72466, sequence calculation, excluding the transit peptide, x * 85000, about, recombinant SBD-fusion enzyme, SDS-PAGE
85000
x * 72466, sequence calculation, excluding the transit peptide, x * 85000, about, recombinant SBD-fusion enzyme, SDS-PAGE
70000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 72466, sequence calculation, excluding the transit peptide, x * 85000, about, recombinant SBD-fusion enzyme, SDS-PAGE
additional information
additional information
three-dimensional structure analysis. Structure comparison with the amylosucrase from Deinococcus geothermalis, overview
additional information
-
three-dimensional structure analysis. Structure comparison with the amylosucrase from Deinococcus geothermalis, overview
additional information
the active pocket in the enzyme is surrounded by A, B, and B' domains. The A domain serves as a catalytic domain andhas the typical (beta/alpha)8-barrel structure of the GH13 family. The B' domain includes oligosaccharide binding sites, and both the B and B' domains of the enzyme contain acceptor subsites (+2, +3, +4, +5, and +6)
additional information
-
the active pocket in the enzyme is surrounded by A, B, and B' domains. The A domain serves as a catalytic domain andhas the typical (beta/alpha)8-barrel structure of the GH13 family. The B' domain includes oligosaccharide binding sites, and both the B and B' domains of the enzyme contain acceptor subsites (+2, +3, +4, +5, and +6)
additional information
the residues forming subsites -1 and +1 are considered to be likely involved in the activity as well as the overall stability of the enzyme, active site structure, overview
additional information
-
the residues forming subsites -1 and +1 are considered to be likely involved in the activity as well as the overall stability of the enzyme, active site structure, overview
additional information
-
amylosucrases adopt a deep pocket topology of about 15 A with the catalytic triad located at the bottom
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
cocrystallization of E328Q mutant enzyme with maltoheptaose, X-ray structure at 2.2 A resolution
purified recombinant detagged NpAS in complex with turanose, hanging drop vapor diffusion method, 1:1 v/v ratio of protein, containing 6 mg/ml in 20 mM Tris, pH 8.0, to precipitant solution containing 1.5 M sodium acetate, 0.1 M sodium cacodylate, pH 7.0, 2 weeks, X-ray diffraction structure determination and analysis at 1.85 A resolution
recombinant enzyme, equal amounts of 4 mg/ml enzyme in 150 mM NaCl, 50 mM Tris-HCl, pH 7.0, 1 mM EDTA and 1 mM dithiothreitol and reservoir solution consisting of 30% polyethylene glycol 6000 and 100 mM HEPES, pH 7.0, crystal structure at 1.4 A resolution
the mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A is crystallized by hanging drop vapor diffusion method at 12°C
crystal structure of the acid/base catalyst mutant, E328Q, with a covalently bound glucopyranosyl moiety. The structure is refined to a resolution of 2.2 A and shows that binding of the covalent intermediate results in a backbone movement of 1 A around the location of the nucleophile, Asp286
-
crystal structure of wild-type amylosucrase in complex with beta-D-glucose at 1.66 A, crystal structure of E328Q mutant enzyme in complex with sucrose at 2.0 A resolution
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E227G
mutant enzyme is a highly efficient polymerase that produces a longer polymer than the wild-type enzyme. Decreased stability and the temperature optimum compared to wild-type enzyme
E328Q
N387D
60% increase in activity compared to wild-type enzyme, increased stability at 50°C
R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D
compared to the wild-type enzyme, and in agreement with the loss of polymerase activity, the mutant enzyme incorporates higher amounts of glucosyl units in maltose (24.8% versus 5.8% for wild-type enzyme) and in maltotriose (18.6% versus 2.9% for wild-type enzyme). The mutant enzyme incorporates 23% glucosyl units into erlose (alpha-D-glucopyranosyl-(1->4)-alpha-D-glucopyranosyl-(1->2)-beta-D-fructose). No glucosyl units are incorporated into erlose (alpha-D-glucopyranosyl-(1->4)-alpha-D-glucopyranosyl-(1->2)-beta-D-fructose)by the wild-type enzyme. Panose (alpha-D-glucopyranosyl-(1->6)-alpha-D-glucopyranosyl-(1->4)-alpha-D-glucose) is produced by the mutant enzyme, 13.9% of the glucosyl units are incorporated into this trisaccharide. No panose is produced by the wild-type enzyme. The Tm-value is slightly lowered compared to wild-type enzyme (-3°C)
R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R
compared to the wild-type enzyme, and in agreement with the loss of polymerase activity, the mutant enzyme incorporates higher amounts of glucosyl units in maltose (15.3% versus 5.8% for wild-type enzyme) and in maltotriose (23% versus 2.9% for wild-type enzyme). The mutant enzyme incorporates nearly 46% of the glucosyl residues in turanose, versus only 19% for the wild-type enzyme. Panose (alpha-D-glucopyranosyl-(1->6)-alpha-D-glucopyranosyl-(1->4)-alpha-D-glucose) is produced by the mutant enzyme, 8.5% of the glucosyl units are incorporated into this trisaccharide. No panose is produced by the wild-type enzyme. The Tm-value is slightly lowered compared to wild-type enzyme (-1.9°C)
R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A
mutant enzyme that shows a 6fold enhanced activity toward sucrose compared to the wild-type enzyme. Only soluble maltooligosaccharide products of controlled size chains (2 < DP < 21) with a narrow polydispersity are observed. This variant, containing 9 mutations in the active site, is characterized at both biochemical and structural levels. Its X-ray structure is determined and further investigated by molecular dynamics to understand the molecular origins of its higher activity on sucrose and higher production of small maltooligosaccharides, with a totally abolished insoluble glucan synthesis
R226L/I228V/F229A/A289I/F290Y/E300I/V331T
compared to the wild-type enzyme, and in agreement with the loss of polymerase activity, the mutant enzyme incorporates higher amounts of glucosyl units in maltose (27.3% versus 5.8% for wild-type enzyme) and in maltotriose (20.5% versus 2.9% for wild-type enzyme). With the mutant enzyme 20% glucosyl units are incorporated into erlose (alpha-D-glucopyranosyl-(1->4)-alpha-D-glucopyranosyl-(1->2)-beta-D-fructose). No glucosyl units are incorporated into erlose by the wild-type enzyme. 1.6% panose is produced by the mutants enzyme and it is not produced by the wild-type enzyme. The Tm-value is slightly lowered compared to wild-type enzyme (-3°C)
D394A
-
23.5% of the wild-type activity, according to the initial rate of sucrose consumption, very poor ativation by glycogen
R226A
-
activated by the products it forms. The mutant yields twice as much insoluble glucan as the wild-type enzyme and leads to the production of lower quantities of by-products, mutant enzyme is strongly activated by glycogen
R226N
-
site-directed mutagenesis, compared to the wild-type enzyme, the mutant shows a 10fold enhancement in the catalytic efficiency and a nearly twofold higher production of an insoluble amylose-like polymer
R226X
-
site-directed mutagenesis, the single site mutants, except R226N, show reduced activity compare to the wild-type enzyme
R415A
-
4.3% of the activity compared with the wild-type enzyme. No synthesis of any insoluble modified glycogen
R446A
-
15% of the wild-type activity, according to the initial rate of sucrose consumption, no synthesis of any insoluble modified glycogen
synthesis
-
potential use for the synthesis or the modification of polysaccharides is limited by its low catalytic efficiency on sucrose alone, its low stability, and its side reactions resulting in sucrose isomer formation. Development of a zero background expression cloning strategy for the generation of large variant libraries, a selection mechanism to discard inactive variants, and a screening method for identification of interesting clones
additional information
E328Q
inactive mutant, sucrose binding structure analysis using the crystal structure, PDB ID 1JGI
additional information
random mutagenesis, high-throughput screening and isolation of amylosucrase variants displaying higher thermostability or increased resistance to organic solvents, overview
additional information
generation of amylosucrase variants that terminate catalysis of acceptor elongation at the di- or trisaccharide stage, product patterns formed by wild-type enzyme and selected genetic variants in the presence of sucrose as the sole substrate, overview
additional information
amylosucrase from Neisseria polysaccharea is fused to a starch-binding domain (SBD) of cyclodextrin glycosyltransferase from Bacillus circulans, expression of the amylosucrase-SBD and SBD-amylosucrase fusion proteins in the amylose-containing (cv. Kardal) and amylose-free (amf) Solanum tuberosum plants, respectively. Expression of SBD-amylosucrase fusion protein in the amylose-containing potato results in starch granules with a rough surface, a twofold increase in median granule size, and altered physico-chemical properties including improved freeze-thaw stability, higher end viscosity, and better enzymatic digestibility. These effects are possibly a result of the physical interaction between amylosucrase and starch granules
additional information
-
amylosucrase from Neisseria polysaccharea is fused to a starch-binding domain (SBD) of cyclodextrin glycosyltransferase from Bacillus circulans, expression of the amylosucrase-SBD and SBD-amylosucrase fusion proteins in the amylose-containing (cv. Kardal) and amylose-free (amf) Solanum tuberosum plants, respectively. Expression of SBD-amylosucrase fusion protein in the amylose-containing potato results in starch granules with a rough surface, a twofold increase in median granule size, and altered physico-chemical properties including improved freeze-thaw stability, higher end viscosity, and better enzymatic digestibility. These effects are possibly a result of the physical interaction between amylosucrase and starch granules
additional information
construction of chimeric enzymes using gene dgas and gene npas from Deinococcus geothermalis by overlap extension polymerase chain reaction, the mutants show altered polymerization activity and thermostability
additional information
-
construction of chimeric enzymes using gene dgas and gene npas from Deinococcus geothermalis by overlap extension polymerase chain reaction, the mutants show altered polymerization activity and thermostability
additional information
introduction of mutations at D144, Y147, F250, R284, and R509 positions leads to equivalent or impaired stability compared with the wild-type enzyme. Several mutant variants retain their transglucosylation activity and are still able to catalyze the synthesis of maltooligosaccharides. In particular, two mutants H392P and Y147F display original and controlled product distributions compared to the wild-type parental NpAS being more efficient for synthesizing soluble oligosaccharides. Two H187 variants and nine H392 variants show lower free energy values than that calculated for the wild-type enzyme
additional information
-
introduction of mutations at D144, Y147, F250, R284, and R509 positions leads to equivalent or impaired stability compared with the wild-type enzyme. Several mutant variants retain their transglucosylation activity and are still able to catalyze the synthesis of maltooligosaccharides. In particular, two mutants H392P and Y147F display original and controlled product distributions compared to the wild-type parental NpAS being more efficient for synthesizing soluble oligosaccharides. Two H187 variants and nine H392 variants show lower free energy values than that calculated for the wild-type enzyme
additional information
-
non-covalent immobilization of the recombinant enzyme for use as biocatalyst, about 87% of enzyme activity and 96% of protein are recovered after immobilization, repeated catalysis with acceptable stability, significantly improved thermostability at 40°C compared to the native enzyme, and unaltered temperature and pH profiles, overview
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
43.2 - 51.5
the half-lives of mutant NPAS-B' and wild-type NPAS are 4.28 and 9.99 min at 45°C and their melting temperatures are 43.25°C and 51.52°C, respectively
46
46.1
Tm-value, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A
47.1
Tm-value, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437S/N439D/C445R
49
50
additional information
46
Tm-value, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/G396S/T398V/Q437R/N439D
46
Tm-value, mutant enzyme R226L/I228V/F229A/A289I/F290Y/E300I/V331T
additional information
the quaternary organization is likely to participate in the enhanced thermal stability of the protein
additional information
-
the quaternary organization is likely to participate in the enhanced thermal stability of the protein
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant GST-tagged NpAS by glutathione affinity chromatography, followed by proteolytic removal of the GST-tag
recombinant His-tagged NPAS from Escherichia coli strain BL21 by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloned into an inducible expression system in Escherichia coli
DNA library construction and DNA sequence determination and analysis, expression of the His-tagged enzyme in Escherichia coli
expression of wild-type and random mutants in Escherichia coli strain JM109
gene ams, recombinant expression as starch-binding domain-fusion protein in Solanum tuberosum plants, cv. Kardal and amf, using the Agrobacterium tumefaciens transformation method
gene npas, recombinant expression of C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
recombinant expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain JM109
potential use for the synthesis or the modification of polysaccharides is limited by its low catalytic efficiency on sucrose alone, its low stability, and its side reactions resulting in sucrose isomer formation. Development of a zero background expression cloning strategy for the generation of large variant libraries, a selection mechanism to discard inactive variants, and a screening method for identification of interesting clones
-
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain JM109
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
drug development
amylosucrase has great potential in the biotechnology and food industries, due to its multifunctional enzyme activities. It can synthesize alpha-1,4-glucans, like amylose, from sucrose as a sole substrate. It can also utilize various other molecules as acceptors. In addition, amylosucrase produces sucrose isomers such as turanose and trehalulose. It also efficiently synthesizes modified starch with increased ratios of slow digestive starch and resistant starch, and glucosylated functional compounds with increased water solubility and stability. It produces turnaose more efficiently than other carbohydrate-active enzymes. Amylose synthesized by amylosucrase forms microparticles and these can be utilized as biocompatible materials with various bio-applications, including drug delivery, chromatography, and bioanalytical sciences
food industry
industry
cyclodextrins are frequently utilized chemical substances in the food, pharmaceutical, cosmetics, and chemical industries. An enzymatic process for cyclodextrin production is developed by utilizing sucrose as raw material instead of corn starch. Cyclodextrin glucanotransferase from Paenibacillus macerans is applied to produce the cyclodextrins from linear alpha-(1,4)-glucans, which are obtained by Neisseria polysaccharea amylosucrase treatment on sucrose. The greatest cyclodextrin yield (21.1%, w/w) is achieved from a one-pot dual enzyme reaction at 40°C for 24 h. The maximum level of cyclodextrin production (15.1 mg/ml) is achieved with 0.5 M sucrose in a simultaneous mode of dual enzyme reaction, whereas the reaction with 0.1 M sucrose is the most efficient with regard to conversion yield. Dual enzyme synthesis of cyclodextrins is successfully carried out with no need of starch material. Efficient bioconversion process that does not require the high temperature necessary for starch liquefaction by thermostable alpha-amylase in conventional industrial processing
synthesis
biotechnology
-
treatment of pre-gelatinized rice and barley starches with amylosucrase from Neisseria polysaccharea is a potential way of replacing commercial resistant starch production
synthesis
biotechnology
the enzyme fused to a starch-binding domain (SBD) is introduced in two potato genetic backgrounds to synthesize starch granules with altered composition, and thereby to broaden starch applications. The modified larger starches not only have great benefit to the potato starch industry by reducing losses during starch isolation, but also have an advantage in many food applications such as frozen food due to its extremely high freeze-thaw stability. Modified starches show a higher digestibility after alpha-amylase treatment
biotechnology
amylosucrase has great potential in the biotechnology and food industries, due to its multifunctional enzyme activities. It can synthesize alpha-1,4-glucans, like amylose, from sucrose as a sole substrate. It can also utilize various other molecules as acceptors. In addition, amylosucrase produces sucrose isomers such as turanose and trehalulose. It also efficiently synthesizes modified starch with increased ratios of slow digestive starch and resistant starch, and glucosylated functional compounds with increased water solubility and stability. It produces turnaose more efficiently than other carbohydrate-active enzymes. Amylose synthesized by amylosucrase forms microparticles and these can be utilized as biocompatible materials with various bio-applications, including drug delivery, chromatography, and bioanalytical sciences
food industry
amylosucrase has great potential in the biotechnology and food industries, due to its multifunctional enzyme activities. It can synthesize alpha-1,4-glucans, like amylose, from sucrose as a sole substrate. It can also utilize various other molecules as acceptors. In addition, amylosucrase produces sucrose isomers such as turanose and trehalulose. It also efficiently synthesizes modified starch with increased ratios of slow digestive starch and resistant starch, and glucosylated functional compounds with increased water solubility and stability. It produces turnaose more efficiently than other carbohydrate-active enzymes. Amylose synthesized by amylosucrase forms microparticles and these can be utilized as biocompatible materials with various bio-applications, including drug delivery, chromatography, and bioanalytical sciences
food industry
cyclodextrins are frequently utilized chemical substances in the food, pharmaceutical, cosmetics, and chemical industries. An enzymatic process for cyclodextrin production is developed by utilizing sucrose as raw material instead of corn starch. Cyclodextrin glucanotransferase from Paenibacillus macerans is applied to produce the cyclodextrins from linear alpha-(1,4)-glucans, which are obtained by Neisseria polysaccharea amylosucrase treatment on sucrose. The greatest cyclodextrin yield (21.1%, w/w) is achieved from a one-pot dual enzyme reaction at 40°C for 24 h. The maximum level of cyclodextrin production (15.1 mg/ml) is achieved with 0.5 M sucrose in a simultaneous mode of dual enzyme reaction, whereas the reaction with 0.1 M sucrose is the most efficient with regard to conversion yield. Dual enzyme synthesis of cyclodextrins is successfully carried out with no need of starch material. Efficient bioconversion process that does not require the high temperature necessary for starch liquefaction by thermostable alpha-amylase in conventional industrial processing
food industry
the study investigates the differences in structural and physicochemical properties, especially contents of resistant starch, between native and acid-thinned waxy corn starches treated with amylosucrase from Neisseria polysaccharea. The enzyme exhibits similar catalytic efficiency for both forms of starch. The modified starches have higher proportions of long (DP > 33) and intermediate chains (DP 13-33), and X-ray diffraction showesa B-type crystalline structure for all modified starches. With increasing reaction time, the relative crystallinity and endothermic enthalpy of the modified starches gradually decreases, whereas the melting peak temperatures and resistant starch contents increases. Slight differences are observed in thermal parameters, relative crystallinity, and branch chain length distribution between the modified native and acid-thinned starches. The digestibility of the modified starches is not affected by acid hydrolysis pretreatment, but is affected by the percentage of intermediate and long chains
synthesis
the enzyme can efficiently be used for synthesis of salicin glycosides with sucrose serving as the glucopyranosyl donor and salicin as the acceptor molecule
synthesis
the enzyme might be useful for important tailoring reactions for the generation of bioactive compounds by glycosylation
synthesis
amylosucrase has great potential in the biotechnology and food industries, due to its multifunctional enzyme activities. It can synthesize alpha-1,4-glucans, like amylose, from sucrose as a sole substrate. It can also utilize various other molecules as acceptors. In addition, amylosucrase produces sucrose isomers such as turanose and trehalulose. It also efficiently synthesizes modified starch with increased ratios of slow digestive starch and resistant starch, and glucosylated functional compounds with increased water solubility and stability. It produces turnaose more efficiently than other carbohydrate-active enzymes. Amylose synthesized by amylosucrase forms microparticles and these can be utilized as biocompatible materials with various bio-applications, including drug delivery, chromatography, and bioanalytical sciences
synthesis
mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A only produces soluble oligosaccharides as no insoluble high molecular weight amylose is observed. The mutant enzyme is an attractive enzymatic tool that could offer interesting opportunities for the design of amylodextrins with controlled size
synthesis
-
mutant enzyme R226A, that is activated by the products it forms and yields twice as much insoluble glucan and lower quantities of by-products as the wild-type enzyme is a very promising enzyme for industrial synthesis of amylose-like polymers
synthesis
-
potentiality of amylosucrase in the design of amylodextrins with controlled morphology, structure, and physicochemical properties
synthesis
-
potential of amylosucrase in the design of original carbohydrate-based dendritic nanoparticles
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Remaud-Simeon, M.; Albaret, F.; Canard, B.; Varlet, I.; Colonna, P.; Willemot, R.M.; Monsan, P.
Studies on a recombinant amylosucrase
Prog. Biotechnol.
10
313-320
1995
Neisseria polysaccharea
-
De Montalk, G.P.; Remaud-Simeon, M.; Willemot, R.M.; Planchot, V.; Monsan, P.
Sequence analysis of the gene encoding amylosucrase from Neisseria polysaccharea and characterization of the recombinant enzyme
J. Bacteriol.
181
375-381
1999
Neisseria polysaccharea
Potocki de Montalk, G.; Remaud-Simeon, M.; Willemot, R.M.; Sarcabal, P.; Planchot, V.; Monsan, P.
Amylosucrase from Neisseria polysaccharea: novel catalytic properties
FEBS Lett.
471
219-223
2000
Neisseria polysaccharea
Potocki de Montalk, G.; Remaud-Simeon, M.; Willemot, R.M.; Monsan, P.
Characterization of the activator effect of glycogen on amylosucrase from Neisseria polysaccharea
FEMS Microbiol. Lett.
186
103-108
2000
Neisseria polysaccharea
Skov, L.K.; Mirza, O.; Henriksen, A.; de Montalk, G.P.; Remaud-Simeon, M.; Sarcabal, P.; Willemot, R.M.; Monsan, P.; Gajhede, M.
Amylosucrase, a glucan-synthesizing enzyme from the alpha-amylase family
J. Biol. Chem.
276
25273-25278
2001
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Skov, L.K.; Mirza, O.; Sprogoe, D.; Dar, I.; Remaud-Simeon, M.; Albenne, C.; Monsan, P.; Gajhede, M.
Oligosaccharide and Sucrose Complexes of Amylosucrase
J. Biol. Chem.
277
47741-47747
2002
Neisseria polysaccharea (Q9ZEU2)
Mirza, O.; Skov, L.K.; Remaud-Simeon, M.; Potocki de Montalk, G.; Albenne, C.; Monsan, P.; Gajhede, M.
Crystal structures of amylosucrase from Neisseria polysaccharea in complex with D-glucose and the active site mutant Glu328Gln in complex with the natural substrate sucrose
Biochemistry
40
9032-9039
2001
Neisseria polysaccharea
Albenne, C.; Potocki De Montalk, G.; Monsan, P.; Skov, L.; Mirza, O.; Gajhede, M.; Remaud-Simeon, M.
Site-directed mutagenesis of key amino acids in the active site of amylosucrase from Neisseria polysaccharea
Biologia (Bratisl. )
57
119-128
2002
Neisseria polysaccharea
-
Jensen, M.H.; Mirza, O.; Albenne, C.; Remaud-Simeon, M.; Monsan, P.; Gajhede, M.; Skov, L.K.
Crystal structure of the covalent intermediate of amylosucrase from Neisseria polysaccharea
Biochemistry
43
3104-3110
2004
Neisseria polysaccharea
Potocki-Veronese, G.; Putaux, J.L.; Dupeyre, D.; Albenne, C.; Remaud-Simeon, M.; Monsan, P.; Buleon, A.
Amylose synthesized in vitro by amylosucrase: morphology, structure, and properties
Biomacromolecules
6
1000-1011
2005
Neisseria polysaccharea
van der Veen, B.A.; Potocki-Veronese, G.; Albenne, C.; Joucla, G.; Monsan, P.; Remaud-Simeon, M.
Combinatorial engineering to enhance amylosucrase performance: construction, selection, and screening of variant libraries for increased activity
FEBS Lett.
560
91-97
2004
Neisseria polysaccharea
Albenne, C.; Skov, L.K.; Mirza, O.; Gajhede, M.; Feller, G.; D'Amico, S.; Andre, G.; Potocki-Veronese, G.; van der Veen, B.A.; Monsan, P.; Remaud-Simeon, M.
Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase
J. Biol. Chem.
279
726-734
2004
Neisseria polysaccharea
Rolland-Sabate, A.; Colonna, P.; Potocki-Veronese, G.; Monsan, P.; Planchot, V.
Elongation and insolubilisation of ?-glucans by the action of Neisseria polysaccharea amylosucrase.
J. Cereal Sci.
40
17-30
2004
Neisseria polysaccharea
Putaux, J.L.; Potocki-Veronese, G.; Remaud-Simeon, M.; Buleon, A.
alpha-D-Glucan-based dendritic nanoparticles prepared by in vitro enzymatic chain extension of glycogen
Biomacromolecules
7
1720-1728
2006
Neisseria polysaccharea
van der Veen, B.A.; Skov, L.K.; Potocki-Veronese, G.; Gajhede, M.; Monsan, P.; Remaud-Simeon, M.
Increased amylosucrase activity and specificity, and identification of regions important for activity, specificity and stability through molecular evolution
FEBS J.
273
673-681
2006
Neisseria polysaccharea (Q9ZEU2)
Albenne, C.; Skov, L.K.; Tran, V.; Gajhede, M.; Monsan, P.; Remaud-Simeon, M.; Andre-Leroux, G.
Towards the molecular understanding of glycogen elongation by amylosucrase
Proteins
66
118-126
2007
Neisseria polysaccharea
Emond, S.; Potocki-Veronese, G.; Mondon, P.; Bouayadi, K.; Kharrat, H.; Monsan, P.; Remaud-Simeon, M.
Optimized and automated protocols for high-throughput screening of amylosucrase libraries
J. Biomol. Screen.
12
715-723
2007
Neisseria polysaccharea (Q9ZEU2)
Schneider, J.; Fricke, C.; Overwin, H.; Hofmann, B.; Hofer, B.
Generation of amylosucrase variants that terminate catalysis of acceptor elongation at the di- or trisaccharide stage
Appl. Environ. Microbiol.
75
7453-7460
2009
Neisseria polysaccharea (Q9ZEU2)
Jung, J.H.; Seo, D.H.; Ha, S.J.; Song, M.C.; Cha, J.; Yoo, S.H.; Kim, T.J.; Baek, N.I.; Baik, M.Y.; Park, C.S.
Enzymatic synthesis of salicin glycosides through transglycosylation catalyzed by amylosucrases from Deinococcus geothermalis and Neisseria polysaccharea
Carbohydr. Res.
344
1612-1619
2009
Deinococcus geothermalis, Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Wang, R.; Kim, J.H.; Kim, B.S.; Park, C.S.; Yoo, S.H.
Preparation and characterization of non-covalently immobilized amylosucrase using a pH-dependent autoprecipitating carrier
Biores. Technol.
102
6370-6374
2011
Neisseria polysaccharea, Neisseria polysaccharea ATCC 43768
Kim, J.H.; Wang, R.; Lee, W.H.; Park, C.S.; Lee, S.; Yoo, S.H.
One-pot synthesis of cycloamyloses from sucrose by dual enzyme treatment: combined reaction of amylosucrase and 4-alpha-glucanotransferase
J. Agric. Food Chem.
59
5044-5051
2011
Neisseria polysaccharea
Guerin, F.; Barbe, S.; Pizzut-Serin, S.; Potocki-Veronese, G.; Guieysse, D.; Guillet, V.; Monsan, P.; Mourey, L.; Remaud-Simeon, M.; Andre, I.; Tranier, S.
Structural investigation of the thermostability and product specificity of amylosucrase from the bacterium Deinococcus geothermalis
J. Biol. Chem.
287
6642-6654
2012
Deinococcus geothermalis, Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Cambon, E.; Barbe, S.; Pizzut-Serin, S.; Remaud-Simeon, M.; Andre, I.
Essential role of amino acid position 226 in oligosaccharide elongation by amylosucrase from Neisseria polysaccharea
Biotechnol. Bioeng.
111
1719-1728
2014
Neisseria polysaccharea
Kim, B.S.; Kim, H.S.; Yoo, S.H.
Characterization of enzymatically modified rice and barley starches with amylosucrase at scale-up production
Carbohydr. Polym.
125
61-68
2015
Neisseria polysaccharea
Daude, D.; Champion, E.; Morel, S.; Guieysse, D.; Remaud-Simeon, M.; Andre, I.
Probing substrate promiscuity of amylosucrase from Neisseria polysaccharea
ChemCatChem
5
2288-2295
2013
Neisseria polysaccharea (Q9ZEU2)
-
Seo, D.H.; Jung, J.H.; Jung, D.H.; Park, S.; Yoo, S.H.; Kim, Y.R.; Park, C.S.
An unusual chimeric amylosucrase generated by domain-swapping mutagenesis
Enzyme Microb. Technol.
86
7-16
2016
Deinococcus geothermalis (Q1J0W0), Deinococcus geothermalis, Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Kim, B.K.; Kim, H.I.; Moon, T.W.; Choi, S.J.
Branch chain elongation by amylosucrase: production of waxy corn starch with a slow digestion property
Food Chem.
152
113-120
2014
Neisseria polysaccharea (Q9ZEU2)
Overwin, H.; Wray, V.; Hofer, B.
Biotransformation of phloretin by amylosucrase yields three novel dihydrochalcone glucosides
J. Biotechnol.
211
103-106
2015
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea, Neisseria polysaccharea ATCC 43768 (Q9ZEU2)
Huang, X.F.; Nazarian-Firouzabadi, F.; Vincken, J.P.; Ji, Q.; Visser, R.G.; Trindade, L.M.
Expression of an amylosucrase gene in potato results in larger starch granules with novel properties
Planta
240
409-421
2014
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Daude, D.; Topham, C.M.; Remaud-Simeon, M.; Andre, I.
Probing impact of active site residue mutations on stability and activity of Neisseria polysaccharea amylosucrase
Protein Sci.
22
1754-1765
2013
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Tian, Y.; Xu, W.; Zhang, W.; Zhang, T.; Guang, C.; Mu, W.
Amylosucrase as a transglucosylation tool From molecular features to bioengineering applications
Biotechnol. Adv.
36
1540-1552
2018
Alteromonas macleodii (B6F2H1), Cellulomonas carbonis (A0A0A0BUC7), Cellulomonas carbonis T26 (A0A0A0BUC7), Deinococcus geothermalis (Q1J0W0), Deinococcus geothermalis DSM 11300 (Q1J0W0), Deinococcus radiodurans (Q9RVT9), Deinococcus radiodurans ATCC 13939 (Q9RVT9), Deinococcus radiopugnans, Methylobacillus flagellatus (Q1GY12), Methylotuvimicrobium alcaliphilum (G4T024), Methylotuvimicrobium alcaliphilum DSM 19304 (G4T024), Neisseria polysaccharea (Q9ZEU2), Neisseria subflava (D3A730), Neisseria subflava NJ9703 (D3A730), Pseudarthrobacter chlorophenolicus (B8H6N5), Pseudarthrobacter chlorophenolicus DSM 12829 (B8H6N5), Synechococcus sp. (SMQ77851)
Verges, A.; Barbe, S.; Cambon, E.; Moulis, C.; Tranier, S.; Remaud-Simeon, M.; Andre, I.
Engineering of anp efficient mutant of Neisseria polysaccharea amylosucrase for the synthesis of controlled size maltooligosaccharides
Carbohydr. Polym.
173
403-411
2017
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Zhang, H.; Zhou, X.; He, J.; Wang, T.; Luo, X.; Wang, L.; Wang, R.; Chen, Z.
Impact of amylosucrase modification on the structural and physicochemical properties of native and acid-thinned waxy corn starch
Food Chem.
220
413-419
2017
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
Seo, D.H.; Yoo, S.H.; Choi, S.J.; Kim, Y.R.; Park, C.S.
Versatile biotechnological applications of amylosucrase, a novel glucosyltransferase
Food Sci. Biotechnol.
29
1-16
2020
Alteromonas macleodii (B6F2H1), Alteromonas macleodii KCTC 2957 (B6F2H1), Alteromonas stellipolaris (B6F2G7), Alteromonas stellipolaris KCTC 12195 (B6F2G7), Bifidobacterium thermophilum, Bifidobacterium thermophilum ATCC 25525, Cellulomonas carbonis (A0A0A0BUC7), Cellulomonas carbonis T26 (A0A0A0BUC7), Deinococcus geothermalis (Q1J0W0), Deinococcus geothermalis DSM 11300 (Q1J0W0), Deinococcus radiodurans (Q9RVT9), Deinococcus radiodurans ATCC 13939 (Q9RVT9), Deinococcus radiopugnans (A0A4P8XUU6), Deinococcus radiopugnans ATCC 19172 (A0A4P8XUU6), Methylobacillus flagellatus (Q1GY12), Methylotuvimicrobium alcaliphilum (G4T024), Methylotuvimicrobium alcaliphilum 20Z (G4T024), Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea ATCC 43768 (Q9ZEU2), Neisseria subflava (D3A730), Neisseria subflava ATCC 49275 (D3A730), Pseudarthrobacter chlorophenolicus (B8H6N5), Pseudarthrobacter chlorophenolicus ATCC 700700 (B8H6N5), Synechococcus sp. PCC 7002
Zhang, H.; Zhou, X.; Wang, T.; Luo, X.; Wang, L.; Li, Y.; Wang, R.; Chen, Z.
New insights into the action mode of amylosucrase on amylopectin
Int. J. Biol. Macromol.
88
380-384
2016
Neisseria polysaccharea (Q9ZEU2)
Koh, D.W.; Park, M.O.; Choi, S.W.; Lee, B.H.; Yoo, S.H.
Efficient biocatalytic production of cyclodextrins by combined action of amylosucrase and cyclodextrin glucanotransferase
J. Agric. Food Chem.
64
4371-4375
2016
Neisseria polysaccharea (Q9ZEU2), Neisseria polysaccharea
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