Comparison of different anion-exchange chromatography resins for the purification of cyanobacterial microcystins

2015 ◽  
Vol 16 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Theerasak Somdee ◽  
Anchana Somdee
Keyword(s):  
Liquid Chromatography ◽  
Anion Exchange ◽  
High Performance ◽  
Total Yield ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Ionization Mass ◽  
Freeze Dried ◽  
Wide Range

For the first time, different types of diethylaminoethyl (DEAE) anion-exchange resins, widely used in previous studies, were investigated to determine the most effective resin for the purification of microcystins (MCs). MCs were extracted from freeze-dried Microcystis aeruginosa cells that had been harvested from the Bueng Nong Khot reservoir, Khon Kaen, Thailand. The toxins were precipitated with ammonium sulfate and then fractionated using five different anion-exchange chromatography resins, followed by chromatography with a C18 cartridge. The toxins were further identified via liquid chromatography–electrospray ionization–mass spectrometry (LC-ESI-MS) analysis, and the yields and purity were determined by high-performance liquid chromatography (HPLC) with ultraviolet detection. DEAE Sephadex A-25 exhibited the best overall performance for MC purification regarding both yield and purity, followed by DEAE cellulose, DEAE Sephacel, DEAE Sepharose Fast Flow and Toyopearl DEAE. Four MC variants, MC-RR, MC-FR, [Dha7]MC-LR and MC-WR, were obtained, and [Dha7]MC-LR was the major variant, with a total yield of 53.08 mg and a purity of 95% using the Sephadex resin. This study indicates that protein precipitation and single-column chromatography using DEAE Sephadex A-25 constitute an effective method for the purification of a wide range of MC variants.

Anion-exchange chromatography of proteins on AG MP-1 using high-performance liquid chromatography equipment

1984 ◽  
Vol 142 (2) ◽  
pp. 373-377 ◽  
Author(s):  
John T. Axelson ◽  
James W. Bodley ◽  
Jeou-Yuan Chen ◽  
Patricia C. Dunlop ◽  
Luann P. Rosenthal ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Anion Exchange ◽  
High Performance ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography

Fungal Polygalacturonases Exhibit Different Substrate Degradation Patterns and Differ in Their Susceptibilities to Polygalacturonase-Inhibiting Proteins

1999 ◽  
Vol 12 (8) ◽  
pp. 703-711 ◽  
Author(s):  
B. J. Cook ◽  
R. P. Clay ◽  
C. W. Bergmann ◽  
P. Albersheim ◽  
A. G. Darvill
Keyword(s):  
Anion Exchange ◽  
Plant Species ◽  
High Performance ◽  
Hydrolytic Activity ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Polygalacturonic Acid ◽  
Substrate Degradation ◽  
Wide Range ◽  
Substrate Cleavage

Polygalacturonic acid (PGA) was hydrolyzed by polygalacturonases (PGs) purified from six fungi. The oligogalacturonide products were analyzed by HPAEC-PAD (high performance anion exchange chromatography-pulsed amperimetric detection) to assess their relative amounts and degrees of polymerization. The abilities of the fungal PGs to reduce the viscosity of a solution of PGA were also determined. The potential abilities of four polygalacturonase-inhibiting proteins (PGIPs) from three plant species to inhibit or to modify the hydrolytic activity of the fungal PGs were determined by colorimetric and HPAEC-PAD analyses, respectively. Normalized activities of the different PGs acting upon the same substrate resulted in one of two distinct oligogalacturonide profiles. Viscometric analysis of the effect of PGs on the same substrate also supports two distinct patterns of cleavage. A wide range of susceptibility of the various PGs to inhibition by PGIPs was observed. The four PGs that were inhibited by all PGIPs tested exhibited an endo/exo mode of substrate cleavage, while the three PGs that were resistant to inhibition by one or more of the PGIPs proceed by a classic endo pattern of cleavage.

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