A simple and efficient method for the purification of membrane-bound levansucrase from Zymomonas mobilis

Armands Vigants; Hans Georg Hicke; Stefan Peter Marx

doi:10.1007/s002840010239

A simple and efficient method for the purification of membrane-bound levansucrase from Zymomonas mobilis

Armands Vigants
, Hans Georg Hicke
, Stefan Peter Marx^*

^*Corresponding author for this work

Institute of Microbiology and Biotechnology

Helmholtz-Zentrum Hereon
RWTH Aachen University

Research output: Contribution to journal › Article › peer-review

18 Citations (Scopus)

Abstract

A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilis ATCC 10988 was developed. Levansucrase activity was almost completely recovered and purified by a factor of 15 after precipitation with 0.1 M MnCl₂ as a first capturing step. The enzyme was homogeneously purified by ultrafiltration and anion-exchange chromatography and exhibited a levan-forming activity of 39.2 U mg^-1. The native enzyme formed large aggregates with an apparent molecular mass of more than 10⁶ Da as determined by size-exclusion chromatography, whereas denaturing SDS-PAGE indicated an apparent molecular mass of 50 kDa for the subunits.

Original language	English
Pages (from-to)	415-418
Number of pages	4
Journal	Current Microbiology
Volume	42
Issue number	6
DOIs	https://doi.org/10.1007/s002840010239
Publication status	Published - 2001

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abstract = "A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilis ATCC 10988 was developed. Levansucrase activity was almost completely recovered and purified by a factor of 15 after precipitation with 0.1 M MnCl2 as a first capturing step. The enzyme was homogeneously purified by ultrafiltration and anion-exchange chromatography and exhibited a levan-forming activity of 39.2 U mg-1. The native enzyme formed large aggregates with an apparent molecular mass of more than 106 Da as determined by size-exclusion chromatography, whereas denaturing SDS-PAGE indicated an apparent molecular mass of 50 kDa for the subunits.",

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AU - Vigants, Armands

AU - Hicke, Hans Georg

AU - Marx, Stefan Peter

PY - 2001

Y1 - 2001

N2 - A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilis ATCC 10988 was developed. Levansucrase activity was almost completely recovered and purified by a factor of 15 after precipitation with 0.1 M MnCl2 as a first capturing step. The enzyme was homogeneously purified by ultrafiltration and anion-exchange chromatography and exhibited a levan-forming activity of 39.2 U mg-1. The native enzyme formed large aggregates with an apparent molecular mass of more than 106 Da as determined by size-exclusion chromatography, whereas denaturing SDS-PAGE indicated an apparent molecular mass of 50 kDa for the subunits.

AB - A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilis ATCC 10988 was developed. Levansucrase activity was almost completely recovered and purified by a factor of 15 after precipitation with 0.1 M MnCl2 as a first capturing step. The enzyme was homogeneously purified by ultrafiltration and anion-exchange chromatography and exhibited a levan-forming activity of 39.2 U mg-1. The native enzyme formed large aggregates with an apparent molecular mass of more than 106 Da as determined by size-exclusion chromatography, whereas denaturing SDS-PAGE indicated an apparent molecular mass of 50 kDa for the subunits.

UR - https://www.scopus.com/pages/publications/0035022964

U2 - 10.1007/s002840010239

DO - 10.1007/s002840010239

M3 - Article

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VL - 42

SP - 415

EP - 418

JO - Current Microbiology

JF - Current Microbiology

IS - 6

ER -

A simple and efficient method for the purification of membrane-bound levansucrase from Zymomonas mobilis

Abstract

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