2.4.1.4: amylosucrase
This is an abbreviated version!
For detailed information about amylosucrase, go to the full flat file.
Word Map on EC 2.4.1.4
-
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
- 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
Reaction
Synonyms
AaAS, ACAS, AmAS, AMS, Amy-1, ASASE, BtAS, CcAS, DGAS, DRAS, DRpAS, glucosyltransferase, sucrose-1,4-alpha-glucan, MaAS, MFAS, More, NPAS, NsAS, sucrose-glucan glucosyltransferase, SyAS
ECTree
2.4.1.4 amylosucraseAdvanced search results
results (
results (Engineering
Engineering on EC 2.4.1.4 - amylosucrase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A170V/Q353L
-
random mutagenesis, the mutant shows 3.5fold increased thermostabilty at 50°C compared to the wild-type enzyme
P351S
-
random mutagenesis, the mutant shows 8fold increased thermostabilty at 50°C compared to the wild-type enzyme
R20C/A451T
-
random mutagenesis, the mutant shows 10fold increased thermostabilty at 50°C compared to the wild-type enzyme
D394A
-
23.5% of the wild-type activity, according to the initial rate of sucrose consumption, very poor ativation by glycogen
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
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
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)
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
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
R446A
-
15% of the wild-type activity, according to the initial rate of sucrose consumption, no synthesis of any insoluble modified glycogen
additional information
construction of chimeric enzymes using gene dgas and gene npas from Neisseria polysaccharea by overlap extension polymerase chain reaction, the mutants show altered polymerization activity and thermostability. Three-dimensional modeling structure and molecular dynamics of mutant DGAS-B, quaternary structure
additional information
-
construction of chimeric enzymes using gene dgas and gene npas from Neisseria polysaccharea by overlap extension polymerase chain reaction, the mutants show altered polymerization activity and thermostability. Three-dimensional modeling structure and molecular dynamics of mutant DGAS-B, quaternary structure
additional information
-
random mutagenesis and screening for enzyme variants with increased thermostability, molecular dynamics simulations, overview
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
-
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
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
-
