5.4.99.11: isomaltulose synthase
This is an abbreviated version!
For detailed information about isomaltulose synthase, go to the full flat file.
Word Map on EC 5.4.99.11
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5.4.99.11
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synthesis
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erwinia
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rhapontici
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palatinose
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protaminobacter
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2.4.1.1
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pantoea
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1,4-alpha-d-glucan:orthophosphate
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molasses
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plymuthica
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food industry
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vacuole-targeted
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dispersa
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whiteflies
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bemisia
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honeydew
- 5.4.99.11
- synthesis
- erwinia
- rhapontici
- palatinose
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protaminobacter
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2.4.1.1
- pantoea
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1,4-alpha-d-glucan:orthophosphate
- molasses
- plymuthica
- food industry
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vacuole-targeted
- dispersa
- whiteflies
- bemisia
- honeydew
Reaction
Synonyms
alpha-Glucosyltransferase, alpha-glycosyl transferase, Avin_08330, AZOG, EC 5.4.99.10, esi, isomaltulose synthase, Isomaltulose synthetase, MutB, NX-5, PalI, Pall, PdSIase, SIase, SmuA, Sucrose 6-glucosylmutase, Sucrose alpha-glucosyltransferase, sucrose isomerase, Sucrose mutase, Synthase, isomaltulose, Trehalulose synthase
ECTree
5.4.99.11 isomaltulose synthaseAdvanced search results
results (
results (Engineering
Engineering on EC 5.4.99.11 - isomaltulose synthase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
D241A
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site-directed mutagenesis, glucose binding structure in comparison to the wild-type enzyme by crystal structure analysis
E295Q
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site-directed mutagenesis, sucrose binding structure in comparison to the wild-type enzyme by crystal structure analysis
F297A
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
F321A
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
R325D
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
R328D
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
R335H/R336T/K337I/D338P
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site-directed mutagenesis, glucose binding structure in comparison to the wild-type enzyme by crystal structure analysis
E295Q
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site-directed mutagenesis, sucrose binding structure in comparison to the wild-type enzyme by crystal structure analysis
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F297A
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
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R325D
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
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R328D
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site-directed mutagenesis, the mutant shows an altered product ratio of trehalulose and isomaltulose compared to the wild-type enzyme
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D327N
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turnover number for sucrose: 1.4fold decrease, KM-value for sucrose: 1.65fold increase, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 43.7% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 41.0% increase
D327R
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turnover number for sucrose: 2.45fold decrease, KM-value for sucrose: 2.2fold increase, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 26.5% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 17.7% increase
D329N
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turnover number for sucrose: 1.17fold decrease, KM-value for sucrose: 2.9fold increase, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 52.4% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 54.7% increase
E498P
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temperature-optimum is 40°C compared to 35°C for the wild-type enzyme, maximal specific activity increases by 7%. Half-life at 50°C is 9.45 min compared to 1.81 min for wild-type enzyme. Mutation slightly increases the ratio of turnover number to KM-value. Percent content of monosaccharide decreases from 5.9% of the wild-type enzyme to 3.4%
E498P/R310P
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temperature-optimum is 45°C compared to 35°C for the wild-type enzyme, maximal specific activity increases by 16%. Half-life is 13.61 min compared to 1.81 min for wild-type enzyme. Mutation slightly increases the ratio of turnover number to KM-value. Percent content of monosaccharide decreases from 5.9% of the wild-type enzyme to 3.3%
R325D
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turnover number for sucrose: 14.5fold decrease, KM-value for sucrose: 1.2fold decrease, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 67% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 43.6% increase
R325L
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turnover number for sucrose: 14.5fold decrease, KM-value for sucrose: 1.1fold decrease, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 54.3% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 31.3% increase
R328D
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turnover number for sucrose: 11.9fold decrease, KM-value for sucrose: 2fold increase, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 65.6% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 61.2% increase
R328L
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turnover number for sucrose: 14.5fold decrease, KM-value for sucrose: 5fold increase, alpha-D-glucopyranosyl-1,6-D-fructofuranose content: 51.9% decrease, alpha-D-glucopyranosyl-1,1-D-fructofuranose content: 50.3% increase
D442N
Paraburkholderia acidicola
the mutant favors the transfer reaction with an isomer preference for isomaltulose
R311C
Paraburkholderia acidicola
the mutant demonstrates higher catalytic efficiency for D-glucose production over trehalulose production
D200A
Paraburkholderia acidicola MX-45
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enzyme mutant structure analysis
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D442N
Paraburkholderia acidicola MX-45
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the mutant favors the transfer reaction with an isomer preference for isomaltulose
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E254Q
Paraburkholderia acidicola MX-45
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enzyme mutant structure analysis
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R311C
Paraburkholderia acidicola MX-45
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the mutant demonstrates higher catalytic efficiency for D-glucose production over trehalulose production
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F164L
site-directed mutagenesis, three-dimensional structure from crystal structure analysis, comparison to the wild-type. The mutant shows hydrolytic activity converting sucrose to glucose and fructose, kinetics
R284C
site-directed mutagenesis, three-dimensional structure from crystal structure analysis, comparison to the wild-type. The mutant shows hydrolytic activity converting sucrose to glucose and fructose, kinetics. Presence of Mg2+, Ca2+ and Zn2+ ions and glucose have no effect on the activity of the R284C mutant
E175N
site-directed mutagenesis, the mutant shows increased activity and exhibits an identical pH optimum and a slightly increased optimal temperature (35°C) compared to wild-type enzyme (30°C)
E175N/K576
site-directed mutagenesis, the mutant shows increased activity and exhibits an identical pH optimum and a slightly increased optimal temperature (35°C) compared to wild-type enzyme (30°C)
E428D
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
F297A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F297P
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
F297P/R333K
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
F321A
site-directed mutagenesis, the mutant shows only hydrolytic activity
F321A/F319A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
G176D
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
K174D
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
K576D
site-directed mutagenesis, the mutant shows increased activity and exhibits an identical pH optimum and a slightly increased optimal temperature (35°C) compared to wild-type enzyme (30°C)
N577K
site-directed mutagenesis, the mutant's activity is similar to the wild-type enzyme
R333K
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S575D
site-directed mutagenesis, the mutant's activity is similar to the wild-type enzyme
E175N
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site-directed mutagenesis, the mutant shows increased activity and exhibits an identical pH optimum and a slightly increased optimal temperature (35°C) compared to wild-type enzyme (30°C)
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G176D
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site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
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K174D
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site-directed mutagenesis, the mutant shows reduced activity compared to wild-type
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K576D
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site-directed mutagenesis, the mutant shows increased activity and exhibits an identical pH optimum and a slightly increased optimal temperature (35°C) compared to wild-type enzyme (30°C)
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E428D
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
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F297A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
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F297P
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
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F297P/R333K
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
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additional information
additional information
construction of chimeric mutant enzymes AZF3 and AZF5 from sucrose isomerase AZOG of Azotobacter vinelanddii and and sucrose isomerase, encoded by esi gene, from Enterobacter sp. FMB-1. Mutant AZF3 has the N-terminal region of Enterobacter SIase and C-terminal region of AZOG, while mutant AZF5 contains the N-terminal region of AZOG and C-terminal region of Enterobacter SIase. Determination of transglycosylation activity, overview. Mutant AZF3 shows altered substrate specificity and reduced activity compared to the wild-type enzyme
additional information
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construction of chimeric mutant enzymes AZF3 and AZF5 from sucrose isomerase AZOG of Azotobacter vinelanddii and and sucrose isomerase, encoded by esi gene, from Enterobacter sp. FMB-1. Mutant AZF3 has the N-terminal region of Enterobacter SIase and C-terminal region of AZOG, while mutant AZF5 contains the N-terminal region of AZOG and C-terminal region of Enterobacter SIase. Determination of transglycosylation activity, overview. Mutant AZF3 shows altered substrate specificity and reduced activity compared to the wild-type enzyme
additional information
Azotobacter vinelandii DJ / ATCC BAA-1303 / KCTC 12137
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construction of chimeric mutant enzymes AZF3 and AZF5 from sucrose isomerase AZOG of Azotobacter vinelanddii and and sucrose isomerase, encoded by esi gene, from Enterobacter sp. FMB-1. Mutant AZF3 has the N-terminal region of Enterobacter SIase and C-terminal region of AZOG, while mutant AZF5 contains the N-terminal region of AZOG and C-terminal region of Enterobacter SIase. Determination of transglycosylation activity, overview. Mutant AZF3 shows altered substrate specificity and reduced activity compared to the wild-type enzyme
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additional information
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isomaltulose production via yeast surface display of sucrose isomerase from Enterobacter sp. strain FMB-1 on Saccharomyces cerevisiae strain EBY100, stable at a broad range of temperatures (35-55°C) and pHs (pH 5-7) with optimal temperature and pH at 45°C and pH 7.0, respectively
additional information
construction of chimeric mutant enzymes AZF3 and AZF5 from sucrose isomerase SIase from Enterobacter sp. strain FMB-1 and AZOG of Azotobacter vinelanddii. Mutant AZF3 has the N-terminal region of Enterobacter SIase and C-terminal region of AZOG, while mutant AZF5 contains the N-terminal region of AZOG and C-terminal region of Bacillus halodurans SIase. Gene Avin_08330 encodes the putative sucrose isomerase AZOG in the nitrogen-fixing bacterium Azotobacter vinelandii. The enzyme is a type of pseudo-sucrose isomerase harboring the RLDRD motif, a sucrose isomerase-specific region in 329-333. Neither sucrose isomerization nor hydrolysis activities are observed in recombinant AZOG. Determination of transglycosylation activity, overview. Mutant AZF3 shows altered substrate specificity and reduced activity compared to the wild-type enzyme AZOG. The sucrose is converted into isomaltose, isomatulose and trehalulose, as well as the hydrolysis products glucose and fructose by mutant AZF3, while wild-type AZOG only produces glucose and fructose
additional information
immobilization of Escherichia coli strain BL21(DE3) cells recombinantly expressing the enzyme by sodium alginate and CaCl2 solution in untreated cane molasses and corn steep liquor medium for enzyme production and isomaltulose production
additional information
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mutations of the loop region of enzyme NX-5 result in significant changes of the product ratio between isomaltulose and trehalulose
additional information
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bioinspired production of antibacterial sucrose isomerase-sponge for the synthesis of isomaltulose, enzyme immobilization, method development, optimization, and evaluation, overview. The enzyme is immobilized on a epsilon-poly-L-lysine (epsilons-PL)-gelatin sponge as matrix is produced by the lyophilizing method, using water as a porogen. The carboxyl groups of gelatin are activated by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form reactive NHS esters, which can promote the rate of the synthesis reaction to form a stable amide bond. Then the esters react with amine groups of epsilon-PL to form peptide bonds. Through a series of cross-linking reactions, gelatin and epsilon-PL form a sponge. The affinity of immobilized enzyme SIase to substrate is basically unchanged. Immobilized SIase still exhibits more than 90% sucrose conversion after 13 consecutive cycles, which indicates that it has a good operational stability. Furthermore, the immobilized SIase has the potential for isomaltulose production, with 200 g/l sucrose solution as its substrate in the food industry. Isomaltulose is isolated in 83.58% yield and high purity (97.3%). epsilon-Poly-L-lysine (epsilon-PL), is an ideal carrier for enzyme immobilization and has attracted considerable attention because of its good biocompatibility, antimicrobial activity, and non-toxic characteristic. The loose and porous structures of epsilon-PL-gelatin sponge are critical for ensuring relatively high catalytic efficiency
additional information
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immobilization of cells expressing the recombinant wild-type enzyme, and mutant BN 68069 cells, on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
immobilization of cells expressing the recombinant wild-type enzyme, and mutant BN 68069 cells, on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
immobilization of cells expressing the recombinant wild-type enzyme, and mutant BN 68069 cells, on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
immobilization of cells expressing the recombinant wild-type enzyme, and mutant BN 68069 cells, on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
-
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
-
immobilization of cells expressing the recombinant wild-type enzyme, and mutant BN 68069 cells, on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
-
additional information
-
mutations of the loop region of enzyme NX-5 result in significant changes of the product ratio between isomaltulose and trehalulose
-
additional information
-
bioinspired production of antibacterial sucrose isomerase-sponge for the synthesis of isomaltulose, enzyme immobilization, method development, optimization, and evaluation, overview. The enzyme is immobilized on a epsilon-poly-L-lysine (epsilons-PL)-gelatin sponge as matrix is produced by the lyophilizing method, using water as a porogen. The carboxyl groups of gelatin are activated by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form reactive NHS esters, which can promote the rate of the synthesis reaction to form a stable amide bond. Then the esters react with amine groups of epsilon-PL to form peptide bonds. Through a series of cross-linking reactions, gelatin and epsilon-PL form a sponge. The affinity of immobilized enzyme SIase to substrate is basically unchanged. Immobilized SIase still exhibits more than 90% sucrose conversion after 13 consecutive cycles, which indicates that it has a good operational stability. Furthermore, the immobilized SIase has the potential for isomaltulose production, with 200 g/l sucrose solution as its substrate in the food industry. Isomaltulose is isolated in 83.58% yield and high purity (97.3%). epsilon-Poly-L-lysine (epsilon-PL), is an ideal carrier for enzyme immobilization and has attracted considerable attention because of its good biocompatibility, antimicrobial activity, and non-toxic characteristic. The loose and porous structures of epsilon-PL-gelatin sponge are critical for ensuring relatively high catalytic efficiency
-
additional information
-
immobilization of Escherichia coli strain BL21(DE3) cells recombinantly expressing the enzyme by sodium alginate and CaCl2 solution in untreated cane molasses and corn steep liquor medium for enzyme production and isomaltulose production
-
additional information
-
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
-
additional information
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enzyme immobilization by adsorption on a chromatographic resin, optimal reaction conditions for the conversion of sucrose into isomaltulose catalyzed by the immobilized sucrose isomerase is at a pH range 6.0-6.5 and 35-40°C
additional information
Erwinia sp. D12
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immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
cloning of the PdSIase gene into the expression vector pET28a, functional recombinant expression in Yarrowia lipolytica strain CGMCC7326 on the cell surface using the cell wall protein Pir1 as an anchor protein. Selection via a hygromycin B resistance gene as a selection marker, method, overview
additional information
Pantoea dispersa UQ68
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cloning of the PdSIase gene into the expression vector pET28a, functional recombinant expression in Yarrowia lipolytica strain CGMCC7326 on the cell surface using the cell wall protein Pir1 as an anchor protein. Selection via a hygromycin B resistance gene as a selection marker, method, overview
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additional information
engineering of the enzyme to exhibit isomelezitose synthase activity for production of the probiotically nutrial component, overview
additional information
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engineering of the enzyme to exhibit isomelezitose synthase activity for production of the probiotically nutrial component, overview
additional information
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immobilization of cells expressing the recombinant wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
-
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
additional information
the half-lives of the enzyme mutants E175N, K576D and E175N/K576D are 2.30, 1.78 and 7.65 times greater than that of the wild-type enzyme at 45°C, respectively. The Km values for the E175N, K576D and E175N/K576D mutants decrease by 6.6%, 2.0% and 11.0%, respectively, and their kcat/Km values increase by 38.2%, 4.2% and 19.4%, respectively, compared with those of the wild-type enzyme. After optimizing the conditions for isomaltulose production at 45°C, the E175N, K576D and E175N/K576D mutants display slightly improved isomaltulose yields, compared with the wild-type enzyme. The catalytic efficiencies (kcat/Km values) of E175N, K576D and E175N/K576D are increased by 38.2%, 4.2% and 19.4%, respectively
additional information
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the half-lives of the enzyme mutants E175N, K576D and E175N/K576D are 2.30, 1.78 and 7.65 times greater than that of the wild-type enzyme at 45°C, respectively. The Km values for the E175N, K576D and E175N/K576D mutants decrease by 6.6%, 2.0% and 11.0%, respectively, and their kcat/Km values increase by 38.2%, 4.2% and 19.4%, respectively, compared with those of the wild-type enzyme. After optimizing the conditions for isomaltulose production at 45°C, the E175N, K576D and E175N/K576D mutants display slightly improved isomaltulose yields, compared with the wild-type enzyme. The catalytic efficiencies (kcat/Km values) of E175N, K576D and E175N/K576D are increased by 38.2%, 4.2% and 19.4%, respectively
additional information
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the half-lives of the enzyme mutants E175N, K576D and E175N/K576D are 2.30, 1.78 and 7.65 times greater than that of the wild-type enzyme at 45°C, respectively. The Km values for the E175N, K576D and E175N/K576D mutants decrease by 6.6%, 2.0% and 11.0%, respectively, and their kcat/Km values increase by 38.2%, 4.2% and 19.4%, respectively, compared with those of the wild-type enzyme. After optimizing the conditions for isomaltulose production at 45°C, the E175N, K576D and E175N/K576D mutants display slightly improved isomaltulose yields, compared with the wild-type enzyme. The catalytic efficiencies (kcat/Km values) of E175N, K576D and E175N/K576D are increased by 38.2%, 4.2% and 19.4%, respectively
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additional information
Serratia plymuthica ATCC 15928
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immobilization of cells expressing the recombinant wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
-
additional information
Serratia plymuthica ATCC 15928
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immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
-
additional information
-
immobilization of cells expressing the recombinant wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
-
additional information
-
immobilization of cells expressing the wild-type enzyme on alginate cell pellets for isomaltulose production, overview. The operational stability and enzyme half-life are significantly improved
-
additional information
-
engineering of the enzyme to exhibit isomelezitose synthase activity for production of the probiotically nutrial component, overview
-
