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Literature summary extracted from

  • Goldman, D.; Lavid, N.; Schwartz, A.; Shoham, G.; Danino, D.; Shoham, Y.
    Two active forms of Zymomonas mobilis levansucrase: an ordered microfibril structure of the enzyme promotes levan polymerization (2008), J. Biol. Chem., 283, 32209-32217.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.10 expression in Escherichia coli BL21(DE3) via the T7 RNA polymerase expression system with pET9d Zymomonas mobilis

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.10 H296R no ability of the enzyme to form microfibrils and to synthesize levan at pH 6.0 Zymomonas mobilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.10 sucrose substrate inhibition Zymomonas mobilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.10 additional information
-
 additional information 32 mM (sucrose), 25°C, pH 5.0, total activity Zymomonas mobilis
2.4.1.10 additional information
-
 additional information at pHs higher than 7.0, Zymomonas mobilis levansucrase does not exhibit levan synthesis activity Zymomonas mobilis
2.4.1.10 36
-
 sucrose 25°C, pH 5.0, transfructosylation activity Zymomonas mobilis
2.4.1.10 40
-
 sucrose 25°C, pH 5.0, levan synthesis activity is monitored by following the increase in turbidity at 600 nm that occurrs due to the insoluble levan formation Zymomonas mobilis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.10 additional information
-
 at pH 6.0 apparent MW greater than 1000000 Da Zymomonas mobilis
2.4.1.10 94000
-
 at pH 7.4, purification procedure includes two steps, MnCl2 precipitation and gel filtration, resulting in a 98% pure protein based on SDS-PAGE, calculated MW is 46700 Da, the enzyme appears to be a dimer in solution Zymomonas mobilis

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.10 Zymomonas mobilis Q60114 sacB gene, AAA27695; ATCC10988, expression in Escherichia coli BL21(DE3)
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.10 The cell extract is centrifuged (14,000xg for 15 min) and the soluble supernatant is kept at 4°C. Following the procedure of Vigants, the enzyme is selectively precipitated by adding 0.1 M MnCl2. Next, the solution is centrifuged (14,000xg for 15 min), the pellet is re-suspended in buffer B (50 mM phosphate buffer pH 6.0, 100 mM NaCl and 0.02% sodium azide), and then the soluble protein is passed via a Sephacryl S300 26/60 gel filtration column, running at 1 ml/min with either buffer A at pH 7.4 or with buffer B at pH 6.0. Zymomonas mobilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.10 additional information change in substrate specificity appears to be unique to Zymomonas mobilis levansucrase because the levansucrases from Bacillus subtilis, Rahnella aquatilis, and Lactobacillus reuteri are capable of synthesizing levan in their soluble form and are not reported to form fibril-like structures Zymomonas mobilis ?
-
 ?
2.4.1.10 sucrose + sucrose dimer form Zymomonas mobilis D-glucose + (2,6-beta-D-fructosyl)3 degree of polymerisation 3 ?
2.4.1.10 sucrose + [6)-beta-D-fructofuranosyl-(2->]n alpha-D-glucopyranoside pH lower than 7.0, fibril form Zymomonas mobilis D-glucose + [6)-beta-D-fructofuranosyl-(2->]n+1 alpha-D-glucopyranoside
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.1.10 dimer structure depends on ionic strength and pHs, above 7.0, products are short fructo-saccharides (degree of polymerization 3) Zymomonas mobilis
2.4.1.10 More microfibrils, structure depends on ionic strength and pHs, below 6.0, well-ordered, precipate out of the solution, synthesis of levan (degree of polymerization greater than 20000). The fibril structure decreases the diffusion of the enzyme away from the bacteria cells, which results in the close association of the cell to the high molecular polymer, levan. Zymomonas mobilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.10 379
-
 sucrose transfructosylation activity Zymomonas mobilis
2.4.1.10 411
-
 sucrose levan synthesis activity Zymomonas mobilis
2.4.1.10 461
-
 sucrose total activity Zymomonas mobilis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.10 additional information
-
 Zymomonas mobilis levansucrase exists in two active forms, depending on the pH and ionic strength Zymomonas mobilis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.4.1.10 2000
-
 sucrose
-
 Zymomonas mobilis
2.4.1.10 2100
-
 sucrose
-
 Zymomonas mobilis
2.4.1.10 2800
-
 sucrose
-
 Zymomonas mobilis