Cloned (Comment) | Organism |
---|---|
gene dsrE563, DNA and amino acid sequence determination and analysis, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)pLysS using expression vector pRSETC | Leuconostoc mesenteroides |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of two truncated derivative mutants DsrE563DCD2DGBD (DsrE563-1) and DsrE563DCD2DVR (DsrE563-2). Mutant DsrE563-1 with a deletion of 1620 amino acids from the C-terminus, and mutant DsrE563-2 with deletion of 1258 amino acids from the C-terminus and 349 amino acids from the N-terminus, are catalytically active synthesizing less-soluble dextran, mainly containing alpha-1,6 glucosidic linkage, the synthesized less-soluble dextran also has a branched alpha-1,3 linkage. Mutant DsrE563-2 shows 4.5fold higher dextransucrase activity than mutant DsrE563-1 and a higher acceptor reaction efficiency compared to the wild-type enzyme from Leuconostoc mesenteroides strain 512 FMCM when various mono- or disaccharides are used as acceptors | Leuconostoc mesenteroides |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | required | Leuconostoc mesenteroides |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
132900 | - |
x * 313300, about, sequence calculation, x * 132900, about, mutant DsrE563-1, x * 136800, about, mutant DsrE563-2, sequence calculation, sequence calculation, x * 128000-146000, purified mutant DsrE563-2 exists in multiple molecular forms, SDS-PAGE | Leuconostoc mesenteroides |
136800 | - |
x * 313300, about, sequence calculation, x * 132900, about, mutant DsrE563-1, x * 136800, about, mutant DsrE563-2, sequence calculation, sequence calculation, x * 128000-146000, purified mutant DsrE563-2 exists in multiple molecular forms, SDS-PAGE | Leuconostoc mesenteroides |
313300 | - |
x * 313300, about, sequence calculation, x * 132900, about, mutant DsrE563-1, x * 136800, about, mutant DsrE563-2, sequence calculation, sequence calculation, x * 128000-146000, purified mutant DsrE563-2 exists in multiple molecular forms, SDS-PAGE | Leuconostoc mesenteroides |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
sucrose + [(1->6)-alpha-D-glucosyl]n | Leuconostoc mesenteroides | - |
D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? | |
sucrose + [(1->6)-alpha-D-glucosyl]n | Leuconostoc mesenteroides B-1299CB4 | - |
D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leuconostoc mesenteroides | G8XR50 | constitutive gene dsrE563 | - |
Leuconostoc mesenteroides B-1299CB4 | G8XR50 | constitutive gene dsrE563 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS by anion exchange and hydrophobic interaction chromatography | Leuconostoc mesenteroides |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | dextran product analysis by treatment with Penicillium dextranase and by thin layer chromatography, and by NMR spectroscopy | Leuconostoc mesenteroides | ? | - |
? | |
additional information | dextran product analysis by treatment with Penicillium dextranase and by thin layer chromatography, and by NMR spectroscopy | Leuconostoc mesenteroides B-1299CB4 | ? | - |
? | |
sucrose + D-glucose | - |
Leuconostoc mesenteroides | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | products determined are leucrose, isomaltose, glucosyl leucroside, isomaltotriose, isomaltosyl leucroside, and isomaltotetrose | ? | |
sucrose + D-glucose | - |
Leuconostoc mesenteroides B-1299CB4 | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | products determined are leucrose, isomaltose, glucosyl leucroside, isomaltotriose, isomaltosyl leucroside, and isomaltotetrose | ? | |
sucrose + isomaltose | - |
Leuconostoc mesenteroides | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | products determined are leucroside, glucosyl leucroside, isomaltotriose, and isomaltotetraose | ? | |
sucrose + isomaltose | - |
Leuconostoc mesenteroides B-1299CB4 | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | products determined are leucroside, glucosyl leucroside, isomaltotriose, and isomaltotetraose | ? | |
sucrose + maltose | - |
Leuconostoc mesenteroides | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | products determined are panose, alpha-isomaltosyl-1,6-alpha-D-glucopyranosyl-1,4-D-glucopyranose, and alpha-isomaltotriosyl-1,6-alpha-D-glucopyranosyl-1,4-D-glucopyranose | ? | |
sucrose + maltose | - |
Leuconostoc mesenteroides B-1299CB4 | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | products determined are panose, alpha-isomaltosyl-1,6-alpha-D-glucopyranosyl-1,4-D-glucopyranose, and alpha-isomaltotriosyl-1,6-alpha-D-glucopyranosyl-1,4-D-glucopyranose | ? | |
sucrose + [(1->6)-alpha-D-glucosyl]n | - |
Leuconostoc mesenteroides | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? | |
sucrose + [(1->6)-alpha-D-glucosyl]n | - |
Leuconostoc mesenteroides B-1299CB4 | D-fructose + [(1->6)-alpha-D-glucosyl]n+1 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 313300, about, sequence calculation, x * 132900, about, mutant DsrE563-1, x * 136800, about, mutant DsrE563-2, sequence calculation, sequence calculation, x * 128000-146000, purified mutant DsrE563-2 exists in multiple molecular forms, SDS-PAGE | Leuconostoc mesenteroides |
More | the enzyme contains two catalytic domains (CD1 and CD2). The glucan-binding domain is important for both enzyme expression and dextransucrase activity | Leuconostoc mesenteroides |
Synonyms | Comment | Organism |
---|---|---|
DsrE563 | - |
Leuconostoc mesenteroides |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | - |
assay at | Leuconostoc mesenteroides |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.2 | - |
assay at | Leuconostoc mesenteroides |
General Information | Comment | Organism |
---|---|---|
evolution | sequence comparisons of enzyme DsrE563 and enzyme DsrE, the enzymes show differences in the sequence but similar properties | Leuconostoc mesenteroides |
additional information | the glucan-binding domain is important for both enzyme expression and dextransucrase activity | Leuconostoc mesenteroides |