EC Number | Cloned (Comment) | Organism |
---|---|---|
2.4.1.10 | expression in Escherichia coli | Priestia megaterium |
2.4.1.10 | wild-type enzyme and 16 variants are expressed in Escherichia coli | Priestia megaterium |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.4.1.10 | D257A | site-directed mutagenesis, no activity | Priestia megaterium |
2.4.1.10 | D257A | mutant shows almost no activity | Priestia megaterium |
2.4.1.10 | D95A | site-directed mutagenesis, no activity | Priestia megaterium |
2.4.1.10 | E350A | mutant is nearly inactive | Priestia megaterium |
2.4.1.10 | E350A | site-directed mutagenesis, nearly inactive | Priestia megaterium |
2.4.1.10 | E352A | site-directed mutagenesis, no measureable activity | Priestia megaterium |
2.4.1.10 | L118A | site-directed mutagenesis | Priestia megaterium |
2.4.1.10 | L118A | kcat/Km is 5.5% of wild-type value | Priestia megaterium |
2.4.1.10 | N252A | site-directed mutagenesis | Priestia megaterium |
2.4.1.10 | N252A | kcat/Km is 104% of wild-type value | Priestia megaterium |
2.4.1.10 | R256A | mutant is nearly inactive | Priestia megaterium |
2.4.1.10 | R256A | site-directed mutagenesis, nearly inactive | Priestia megaterium |
2.4.1.10 | R370A | site-directed mutagenesis, after a reaction time of 60 min an accumulation of neokestose (2,6-beta-Fru-betaGlc-1,2-beta-Fru, 32.7 mM) is determined, whereas after 19 h, blastose (2,6-beta-Fru-alpha,betaGlc) is the main reaction product (69.7 mM) | Priestia megaterium |
2.4.1.10 | S173A | site-directed mutagenesis | Priestia megaterium |
2.4.1.10 | S173A | kcat/Km is 46% of wild-type value | Priestia megaterium |
2.4.1.10 | W172A | kcat/Km is 1.4% of wild-type value | Priestia megaterium |
2.4.1.10 | W172A | site-directed mutagenesis, 72fold increase of the KM of the wild-type | Priestia megaterium |
2.4.1.10 | W94A | kcat/Km is 9% of wild-type value | Priestia megaterium |
2.4.1.10 | W94A | site-directed mutagenesis, 9% of the catalytic efficiency of the wild-type | Priestia megaterium |
2.4.1.10 | Y421A | kcat/Km is 1.9% of wild-type value | Priestia megaterium |
2.4.1.10 | Y421A | site-directed mutagenesis, 3%of the wild-type activity | Priestia megaterium |
EC Number | General Stability | Organism |
---|---|---|
2.4.1.10 | in standard reactions, long-term temperature treatment revealed a high protein stability at 37°C for at least 24h | Priestia megaterium |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.4.1.10 | additional information | - |
additional information | 2.3 mM (sucrose) total, mutant enzyme S173A | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 29.2 mM (sucrose) total, mutant enzyme R370A, a 4fold KM value decrease compared with the wild-type enzyme supported the interactions of Arg 370 with the 2-OH and 3-OH of the glucosyl residue in the active site | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 31.9 mM (sucrose) total, mutant enzyme W94A, 9% of the catalytic efficiency of the wild-type | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 35.1 mM (sucrose) total, mutant enzyme N252H | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 4.1 mM (sucrose) total, mutant enzyme N252A | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 480.4 mM (sucrose) total, mutant enzyme W172A, 72fold increase of the KM of the wild-type | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 51.9 mM (sucrose) total, mutant enzyme Y421A, 3%of the wild-type activity | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 6.6 mM (sucrose) total, wild-type, pH 6.6, 37°C, 50 mM Sorensens phosphate buffer, 7.36 mg/l enzyme concentration, reaction time 60 min | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 66.6 mM (sucrose), mutant enzyme L118A | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 7.5 mM (sucrose) total, mutant enzyme N252G | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | 8.4 mM (sucrose) total, mutant enzyme N252D | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | mutant enzyme D257A, no activity | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | mutant enzyme D95A, no activity | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | mutant enzyme E350A, nearly inactive | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | mutant enzyme E352A, no measureable activity | Priestia megaterium | |
2.4.1.10 | additional information | - |
additional information | mutant enzyme R256A, nearly inactive | Priestia megaterium | |
2.4.1.10 | 2.3 | - |
sucrose | pH 6.6, mutant enzyme S173A | Priestia megaterium | |
2.4.1.10 | 4.1 | - |
sucrose | pH 6.6, mutant enzyme N252A | Priestia megaterium | |
2.4.1.10 | 6.6 | - |
sucrose | pH 6.6, wild-type enzyme | Priestia megaterium | |
2.4.1.10 | 31.9 | - |
sucrose | pH 6.6, mutant enzyme W94A | Priestia megaterium | |
2.4.1.10 | 51.9 | - |
sucrose | pH 6.6, mutant enzyme Y421A | Priestia megaterium | |
2.4.1.10 | 66.6 | - |
sucrose | pH 6.6, mutant enzyme L118A | Priestia megaterium | |
2.4.1.10 | 480.4 | - |
sucrose | pH 6.6, mutant enzyme W172A | Priestia megaterium |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
2.4.1.10 | extracellular | - |
Priestia megaterium | - |
- |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.4.1.10 | 52000 | - |
MALDI-TOF | Priestia megaterium |
2.4.1.10 | 52000 | - |
FPLC, proteins are identified by peptide mass fingerprinting, as well as by using postsource decay fragmentation data recorded with an Ultraflex MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) mass spectrometer | Priestia megaterium |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.4.1.10 | Priestia megaterium | - |
DSM319 | - |
2.4.1.10 | Priestia megaterium | - |
strain DSM319, gene sacB, expression in Escherichia coli, 74% identity at the amino acid sequence level with SacB from Bacillus subtilis | - |
2.4.1.10 | Priestia megaterium DSM 319 | - |
DSM319 | - |
2.4.1.10 | Priestia megaterium DSM 319 | - |
strain DSM319, gene sacB, expression in Escherichia coli, 74% identity at the amino acid sequence level with SacB from Bacillus subtilis | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.4.1.10 | - |
Priestia megaterium |
2.4.1.10 | by FPLC using a 15 ml CM-Sepharose column system | Priestia megaterium |
EC Number | Source Tissue | Comment | Organism | Textmining |
---|---|---|---|---|
2.4.1.10 | culture supernatant | - |
Priestia megaterium | - |
EC Number | Storage Stability | Organism |
---|---|---|
2.4.1.10 | -20°C, pH 6.6, stable for more than 6 months | Priestia megaterium |
2.4.1.10 | 20°C, more than 6 months, pH 6.6 | Priestia megaterium |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.4.1.10 | sucrose + (2,6-beta-D-fructosyl)n | polyfructan produced by the levansucrase from Bacillus megaterium has a molecular mass of 2711 kDa and consisted mainly of beta(2,6) linkages. Besides the polyfructan formation, the wild-type levansucrase of Bacillus megaterium also synthesizes five different detectable oligosaccharides. Three products are identified: 1-kestose (isokestose), 6-kestose and nystose which are known acceptors for the transfer of fructosyl units | Priestia megaterium | D-glucose + (2,6-beta-D-fructosyl)n+1 | - |
? | |
2.4.1.10 | sucrose + (2,6-beta-D-fructosyl)n | polyfructan produced by the levansucrase from Bacillus megaterium has a molecular mass of 2711 kDa and consisted mainly of beta(2,6) linkages. Besides the polyfructan formation, the wild-type levansucrase of Bacillus megaterium also synthesizes five different detectable oligosaccharides. Three products are identified: 1-kestose (isokestose), 6-kestose and nystose which are known acceptors for the transfer of fructosyl units | Priestia megaterium DSM 319 | D-glucose + (2,6-beta-D-fructosyl)n+1 | - |
? | |
2.4.1.10 | sucrose + ? | - |
Priestia megaterium | blastose + ? | two-dimensional COSY, TOCSY, HMBC and HSQC experiments | ? | |
2.4.1.10 | sucrose + ? | - |
Priestia megaterium DSM 319 | blastose + ? | two-dimensional COSY, TOCSY, HMBC and HSQC experiments | ? | |
2.4.1.10 | sucrose + ? | - |
Priestia megaterium | polyfructan + ? | polyfructan, molecular mass of 2711 kDa and consisted mainly of beta(2-6) linkages | ? | |
2.4.1.10 | sucrose + ? | - |
Priestia megaterium DSM 319 | polyfructan + ? | polyfructan, molecular mass of 2711 kDa and consisted mainly of beta(2-6) linkages | ? | |
2.4.1.10 | sucrose + sucrose | - |
Priestia megaterium | D-glucose + 1-kestose | identified by HPAEC | ? | |
2.4.1.10 | sucrose + sucrose | - |
Priestia megaterium DSM 319 | D-glucose + 1-kestose | identified by HPAEC | ? | |
2.4.1.10 | sucrose + sucrose | - |
Priestia megaterium | D-glucose + 6-kestose | identified by HPAEC | ? | |
2.4.1.10 | sucrose + sucrose | - |
Priestia megaterium DSM 319 | D-glucose + 6-kestose | identified by HPAEC | ? | |
2.4.1.10 | sucrose + sucrose | - |
Priestia megaterium | D-glucose + neokestose | identified by one-dimensional and correlation spectroscopy (i.e. COSY, TOCSY, HMBC, DEPT and HSQC) | ? | |
2.4.1.10 | sucrose + sucrose | - |
Priestia megaterium | D-glucose + nystose | identified by HPAEC | ? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.4.1.10 | levansucrase | - |
Priestia megaterium |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.4.1.10 | 45 | - |
- |
Priestia megaterium |
2.4.1.10 | 45 | - |
wild-type, pH6.6, 50 mM Sorensens buffer | Priestia megaterium |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.4.1.10 | 37 | - |
high stability for at least 24 h | Priestia megaterium |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.4.1.10 | 325 | - |
sucrose | pH 6.6, mutant enzyme Y421A | Priestia megaterium | |
2.4.1.10 | 335 | - |
sucrose | mutant enzyme Y421A | Priestia megaterium | |
2.4.1.10 | 363 | - |
sucrose | pH 6.6, mutant enzyme S173A | Priestia megaterium | |
2.4.1.10 | 363 | - |
sucrose | mutant enzyme S173A | Priestia megaterium | |
2.4.1.10 | 1000 | - |
sucrose | pH 6.6, mutant enzyme W94A | Priestia megaterium | |
2.4.1.10 | 1000 | - |
sucrose | mutant enzyme W94A | Priestia megaterium | |
2.4.1.10 | 1231 | - |
sucrose | pH 6.6, mutant enzyme L118A | Priestia megaterium | |
2.4.1.10 | 1231 | - |
sucrose | mutant enzyme L118A | Priestia megaterium | |
2.4.1.10 | 1480 | - |
sucrose | pH 6.6, mutant enzyme N252A | Priestia megaterium | |
2.4.1.10 | 1480 | - |
sucrose | mutant enzyme N252A | Priestia megaterium | |
2.4.1.10 | 1529 | - |
sucrose | mutant enzyme N252H | Priestia megaterium | |
2.4.1.10 | 2256 | - |
sucrose | mutant enzyme N252G | Priestia megaterium | |
2.4.1.10 | 2272 | - |
sucrose | wild-type | Priestia megaterium | |
2.4.1.10 | 2272 | - |
sucrose | pH 6.6, wild-type enzyme | Priestia megaterium | |
2.4.1.10 | 2396 | - |
sucrose | pH 6.6, mutant enzyme W172A | Priestia megaterium | |
2.4.1.10 | 2396 | - |
sucrose | mutant enzyme W172A | Priestia megaterium | |
2.4.1.10 | 3743 | - |
sucrose | mutant enzyme N252D | Priestia megaterium |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
2.4.1.10 | 6 | 7 | - |
Priestia megaterium |
2.4.1.10 | 6 | 7 | wild-type, 50 mM Sorensens phosphate buffer at 37°C, containing 500 mM sucrose using an enzyme concentration of 7.36 mg/l | Priestia megaterium |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
2.4.1.10 | 5.6 | 7.6 | the enzyme activity significantly decreases at pH values below 5.6 and above 7.6 | Priestia megaterium |
2.4.1.10 | 5.6 | 7.6 | wild-type, 50 mM Sorensens phosphate buffer at 37°C, containing 500 mM sucrose using an enzyme concentration of 7.36 mg/l | Priestia megaterium |