2.4.1.7: sucrose phosphorylase
This is an abbreviated version!
For detailed information about sucrose phosphorylase, go to the full flat file.
Word Map on EC 2.4.1.7
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2.4.1.7
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mesenteroides
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leuconostoc
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bifidobacterium
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adolescentis
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phosphorylases
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transglucosylation
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synthesis
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alpha-d-glucose
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laminaribiose
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deglucosylation
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dextransucrase
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pseudobutyrivibrio
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ruminis
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kojibiose
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medicine
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industry
- 2.4.1.7
- mesenteroides
- leuconostoc
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bifidobacterium
- adolescentis
- phosphorylases
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transglucosylation
- synthesis
- alpha-d-glucose
- laminaribiose
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deglucosylation
- dextransucrase
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pseudobutyrivibrio
- ruminis
- kojibiose
- medicine
- industry
Reaction
Synonyms
1149SPase, 1355SPase, 742SPase, BiSP, disaccharide glucosyltransferase, LmSPase, More, SPase, sucrose glucosyltransferase, sucrose: orthophosphate, alpha-D-glucosyltransferase, sucrose: phosphate alpha-D-glucosyltransferase, unspase
ECTree
2.4.1.7 sucrose phosphorylaseAdvanced search results
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results (Engineering
Engineering on EC 2.4.1.7 - sucrose phosphorylase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A498H
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site-directed mutagenesis, the mutant shows reduced activity and thermostability compared to the wild-type enzyme
D342A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
D445P
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site-directed mutagenesis, the mutant shows slightly decreased activity and increased thermostability compared to the wild-type enzyme
D445P/D446G
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site-directed mutagenesis, the mutant shows reduced activity and increased thermostability compared to the wild-type enzyme
D445P/D446P
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site-directed mutagenesis, the mutant shows reduced activity and unaltered thermostability compared to the wild-type enzyme
D445P/D446T
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site-directed mutagenesis, the mutant shows reduced activity and increased thermostability compared to the wild-type enzyme
D446G
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site-directed mutagenesis, the mutant shows similar activity and thermostability as the wild-type enzyme
D446P
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site-directed mutagenesis, the mutant shows slightly increased activity and the same thermostability compared to the wild-type enzyme
D446T
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site-directed mutagenesis, the mutant shows reduced activity and slightly reduced thermostability compared to the wild-type enzyme
H234A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
L306H
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site-directed mutagenesis, the mutant shows similar activity and thermostability as the wild-type enzyme
L343A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
N325D/V473H
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site-directed mutagenesis, the mutant shows reduced activity and thermostability compared to the wild-type enzyme
N414D
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site-directed mutagenesis, the mutant shows reduced activity and thermostability compared to the wild-type enzyme
P134A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
Q331E
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site-directed mutagenesis, the mutant shows reduced activity and increased thermostability compared to the wild-type enzyme
Q345A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
Q345F
Q84HQ2
site-directed mutagenesis, the mutation allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97% whereas the wild-type enzyme favours sucrose hydrolysis. The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange, structural changes in the active site of BaSP Q345F, modeling, overview
Q460E/E485H
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site-directed mutagenesis, the mutant shows reduced activity and increased thermostability compared to the wild-type enzyme
R135A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
R393N
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site-directed mutagenesis, the mutant shows reduced activity and increased thermostability compared to the wild-type enzyme
Y132A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
Y196A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
Y344A
Q84HQ2
saturation mutagensis, transglucosylation and hydrolytic side activity of the mutant compared to the wild-type
D196A
D196N/E237Q
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the mutation affects the the stereoselectivity of the reaction
D295E
D295N
D338N
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site-directed mutagenesis of a fructose-binding residue, the mutant shows 7000fold reduced activity compared to the wild-type enzyme due to disruption of steps where fructose departs or attacks
D49A
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site-directed mutagenesis, the mutant enzyme shows 10000000fold reduced enzyme glycosylation and 500fold reduced enzyme deglycosylation compared to the wild-type enzyme. The mutant also shows a loss in selectivity for phosphate against water and substrate inhibition by phosphate
E237Q
F52A
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site-directed mutagenesis, large destabilization of the transition states for enzyme glucosylation and deglucosylation in the mutant compared to the wild-type enzyme, while the relative stability of the glucosyl enzyme intermediate was weakly affected by substitution of Phe52
F52N
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site-directed mutagenesis, large destabilization of the transition states for enzyme glucosylation and deglucosylation in the mutant compared to the wild-type enzyme, while the relative stability of the glucosyl enzyme intermediate was weakly affected by substitution of Phe52
R137A
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site-directed mutagenesis of a phosphate-binding residue, the mutant shows 60fold reduced activity compared to the wild-type enzyme due to disruption of steps where fructose departs or attacks
R395L
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site-directed mutagenesis, the mutant enzyme shows 100000fold reduced enzyme glycosylation and 500fold reduced enzyme deglycosylation compared to the wild-type enzyme. The mutant also shows a loss in selectivity for phosphate against water and substrate inhibition by phosphate
Y340A
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site-directed mutagenesis of a phosphate-binding residue, the mutant shows 2500fold reduced activity compared to the wild-type enzyme due to disruption of steps where fructose departs or attacks
D295E
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site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
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D295N
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site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
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D249G
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
K140M
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
N155S
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
Q144R
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
S62P
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
T47S
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
T47S/S62P/Y77H/V128L/K140M/Q144R/N155S/D249G
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mutant enzyme retains more than 60% of initial activity at 60°C for 20 min
V128L
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
Y77H
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mutation contributes to the enhancement of thermal stability, mutant enzyme retains activity after heat treatment at 55°C for 20 min
additional information
D196A
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inactive mutant enzyme. External azide partly complements the catalytic defect in D196A while formate, acetate and halides can not restore activity. The mutant utilizes azide to convert alpha-D-glucose 1-phosphate into beta-D-glucose 1-azide, reflecting a change in stereochemical course of glucosyl transfer from alpha-retaining in wild-type to inverting in D196A. Phosphorolysis of beta-D-glucose 1-azide by D196A occurrs through a ternary complex kinetic mechanism, in contrast to the wild-type whose reactions feature a common glucosyl enzyme intermediate and ping-pong kinetics
D196A
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site-directed mutagenesis, the purified D196A mutant shows 40% reduced activity compared to the wild-type in phosphorolysis and synthesis of sucrose as well as arsenolysis of alpha-glucose 1-phosphate, however, with azide as an alternative nucleophile, the conversion of alpha-glucose 1-phosphate proceeds at a slow rate and results in the formation of product glucose 1-azide with a beta-anomeric configuration, activity enhancement in the D196A mutant results from the direct participation of azide in the now inverting, single displacement-like mechanism of glucosyl transfer, overview
D295E
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
D295E
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mutation decreases the catalytic center activity of sucrose phosphorylase to about 0.01% of the wild-type level. The 100000fold preference of the wild-type for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost
D295E
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site-directed mutagenesis of the catalytic residue, the mutant shows about 0.01% of the wild-type enzyme activity, the preference of the wild-type enzyme for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost in the mutant enzyme
D295N
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
D295N
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mutation decreases the catalytic center activity of sucrose phosphorylase to about 0.01% of the wild-type level. Glucosylation and deglucosylation steps are affected uniformly, and independently of leaving group ability and nucleophilic reactivity of the substrate, respectively. The 100000fold preference of the wild-type for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost
D295N
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site-directed mutagenesis of the catalytic residue, the mutant shows about 0.01% of the wild-type enzyme activity, the preference of the wild-type enzyme for glucosyl transfer compared with mannosyl transfer from phosphate to fructose is lost in the mutant enzyme
D295N
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site-directed mutagenesis, the mutant shows 0.01% of the wild-type sucrose phosphorolysis activity, meaning a reduction by 20000fold, but regaines activity by heat treatment for 10 min at 100°C, caused by a partial deamidation of D295. The catalytic defect resulting from the substitution of Asp295 is independent of the leaving group ability and nucleophilic reactivity of the substrate
E237Q
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site-directed mutagenesis of the catalytic residue, the mutant shows 0.001% of wild-type enzyme activity, reactions with substrates requiring Broensted catalytic assistance for glucosylation or deglucosylation are selectively slowed at the respective step about 10fold in mutant E237Q compared to the wild-type enzyme. Azide, acetate and formate but not halides restore catalytic activity up to 300fold in E237Q under conditions in which the deglucosylation step is rate-determining
E237Q
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site-directed mutagenesis, the mutant shows altered pH-dependence compared to the wild-type enzyme
E237Q
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site-directed mutagenesis, replacement of the catalytic acid-base Glu237, the mutant does not display hydrolase activity under transglucosylation conditions and therefore provides 7fold enhancement of transfer yield
additional information
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formation of a glutaraldehyde cross-linked enzyme aggregate for high improvement of the enzyme's stability at 60°C, molecular imprinting of the cross-linked enzyme aggregate with a suitable substrate, i.e. glycerol, involving enzyme precipitation by tert-butyl alcohol can 2fold increase the transglucosylation activity, stability and specificity of the modified enzyme, method, overview. The modified enzyme is more useful as industrial biocatalyst than the native enzyme
additional information
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immobilization of the enzyme by cross-linking leads to a 17 degree higher temperature tolerance compared to the soluble enzyme from Bifidobacterium adolescentis, overview
additional information
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increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis, substitution of the most flexible residues with amino acids that occur more frequently at the corresponding positions in related sequences, and substitutions to promote electrostatic interactions
additional information
Q84HQ2
construction of s stabilized enzyme mutant LNFI through 6 point mutations
additional information
Q84HQ2
redesign of the active site of sucrose phosphorylase through a clash-induced cascade of loop shifts
additional information
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redesign of the active site of sucrose phosphorylase through a clash-induced cascade of loop shifts
additional information
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immobilization of the purified recombinant tagged enzyme for continuous production of alpha-D-glucose 1-phosphate from sucrose and phosphate in a packed bed reactor, method optimization, overview
additional information
immobilization of the purified recombinant tagged enzyme for continuous production of alpha-D-glucose 1-phosphate from sucrose and phosphate in a packed bed reactor, method optimization, overview
additional information
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adsorption and enzyme activity of sucrose phosphorylase on lipid Langmuir and Langmuir-Blodgett films with negligible effects on its secondary structure, but providing a favorable environment for preserving the enzyme catalytic activity, attributed to the interaction of the polypeptide structure with the hydrophobic tails of phospholipid dimyristoylphosphatidic acid, thereby facilitating the access of the analyte to the catalytic site of the enzyme, which is ideal for catalyzing the conversion of sucrose to other products, overview
additional information
generation of disrution mutants of sucrose phosphorylase ScrP and sucrose regulator ScrR by double crossover mutagenesis
additional information
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generation of disrution mutants of sucrose phosphorylase ScrP and sucrose regulator ScrR by double crossover mutagenesis
additional information
Limosilactobacillus reuteri LTH5448
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generation of disrution mutants of sucrose phosphorylase ScrP and sucrose regulator ScrR by double crossover mutagenesis
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