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2.4.1.10: levansucrase

This is an abbreviated version!
For detailed information about levansucrase, go to the full flat file.

Word Map on EC 2.4.1.10

Reaction

sucrose
+
[6)-beta-D-fructofuranosyl-(2->]n alpha-D-glucopyranoside
=
D-glucose
 +
[6)-beta-D-fructofuranosyl-(2->]n+1 alpha-D-glucopyranoside

Synonyms

(2,6)-beta-D-fructan:D-glucose 6-fructosyltransferase, 6-SFT, 6G-FFT2, beta-2,6-fructan: D-glucose-1-fructosyltransferase, beta-2,6-fructan:D-glucose 1-fructosyltransferase, beta-2,6-fructosyltransferase, endolevanase, fructansucrase, fructosyltransferase, sucrose 6-, FTF, Lev, LEV-Y, levanase-sucrase, levansucrase, LevB1, LevB1SacB, LevJ, LEVS, LevU, Lsc, Lsc-3, Lsc2, Lsc3, LscA, LscrA, LSD, LsdA, LvnS, m1ft, M1FT protein, SacB, sucrose 6-fructosyltransferase, sucrose: 2, 6-beta-D-fructan 6-beta-Dfructosyltransferase, sucrose:2,6-beta-D-fructan:6-beta-D-fructosyltransferase, sucrose:fructan 6-fructosyltransferase, T1-LS, T2-LS, type 1 levansucrase

ECTree

     2 Transferases
         2.4 Glycosyltransferases
             2.4.1 Hexosyltransferases
                2.4.1.10 levansucrase

Purification

Purification on EC 2.4.1.10 - levansucrase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, DEAE-cellulose column chromatography and Sephadex G-100 gel filtration
-
by applying elastin-like polypeptides tags for simple one-step purification based on temperature-triggered precipitation
-
by FPLC using a 15 ml CM-Sepharose column system
-
CM Toyopearl column chromatography
extracellular and periplasmic forms
-
high ionic strength salt solutions fail to remove the enzyme from the cell wall, e.g. 8 M LiCl, 2 M KCl, 6 M CsCl, 2.5 M ammonium sulfate
-
HisTrap column chromatography
HisTrap column chromatography and Superdex S200 gel filtration
HisTrap purification
-
ion exchange chromatography
native enzyme from strain 8-37-0-1 by ammonium sulfate fractionation, anion-exchange chromatography, and ultrafiltration to homogeneity
-
native enzyme partially from strain NRC33a
-
native extracellular enzyme from cell culture supernatant by ethanol precipitation with 50% v/v ethanol
-
native intracellular enzyme 11.8fold by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
-
Ni-NTA affinity column chromatography
Ni-NTA agarose column chromatography
-
Ni-NTA column
-
Ni-NTA column chromatography
Ni2+-chelating affinity chromatography and Superdex S200 gel filtration
-
Ni2+-chelating affinity column chromatography
one-step purification of affinity-tagged heterologous levansucrase from growth medium
-
partial
partial purification
-
partial purification by acetone precipitation. Full purification by acetone precipitation and DEAE-Sepharose column chromatography
-
partial, PAGE with Tween 80 to avoid aggregation of the enzyme
-
partially, native enzyme forms aggregates
-
precipitation with levan
Aerobacter levanicum
-
process optimization for large scale production
-
purification of affinity tagged Lactobacillus reuteri levansucrase produced by Bacillus megaterium. The gentle one-step purification method for the recombinant protein directly from the cell-free growth medium provides an alternative to Escherichia coli production and purification systems, where cell disruption, several centrifugation steps, and often more than one purification step are necessary. The one-step purification yields comparable amounts of pure and active enzyme
-
recombinant and native wild-type and recombinant mutant from supernatant
recombinant as soluble His-tagged protein, small scale
Q60114
recombinant enzyme from Escherichia coli strain BL21(DE3) by cation exchange chromatography
recombinant from E. coli as His-tagged protein
recombinant from inclusion bodies in E. coli cytoplasm, direct refolding and solubilization with Triton X-100, large scale in fed-batch culture of recombinant E. coli
Q60114
recombinant His-tagged enzyme 4fold by nickel affinity chromatography
-
recombinant His-tagged enzyme from E. coli
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and ultrafiltration
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged LevU by metal affinity chromatography, recombinant LevU fused to a chitin-binding domain from Escherichia coli strain DH5alpha by affinity binding and immobilization on chitin beads
-
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
recombinant isozyme Lsc3 from Escherichia coli by ammonium sulfate fractionation and gel filtration
recombinant overexpression in E. coli, His-tagged protein
-
SDS-PAGE, immobilon-P membrane
-
TALON metal affinity resin column chromatography
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.
Q60114
Toyopearl DEAE-650M column chromatography
-