Salt promoted adsorption chromatography of malted barley amylases

Pazos, C.; Franco-Fraguas, Laura; Batista‐Viera, Francisco

doi:10.1007/bf02290342

Cited by 6 publications

(2 citation statements)

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“…The elution was performed in two steps. The behaviour of wheat [3-amylase towards thiophilic and covalent adsorbents closely resembles the previously reported behaviour of other proteins towards these adsorbents [6,7,8]. The [3-amylase activity was detected only in the first fraction (37 % of the applied enzyme activity, with an E.R.…”

Section: Resultssupporting

confidence: 84%

“…T-gel contains both sulfone and thioether groups. Useful applications of thiophilic adsorbents for the purification of enzymes, including sweet potato 13-amylase [6] and barley malt amylases [7], have been reported. Both T-gel and PyS-gel exhibit strong affinity for immunoglobulins and have proved to be useful tools for their selective purification [3,4].…”

Section: Introductionmentioning

confidence: 99%

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Wheat β-amylase behaviour regarding salt promoted adsorption processes

1998

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The behaviour of wheat [3-amylase from crude extracts in chromatography on agarose gels substituted with different ligand types was investigated. The enzyme displayed high salt promoted adsorption onto thiophilic gels provided with sulfone-thioether and 2-thiopyridine ligands. Quantitative recovery of the enzyme was easily accomplished by elution with buffer in the absence of Na2SO 4. The 3-(2-pyridylthio)-2-hydroxypropylagarose (PyS-gel) also allowed elimination of pigments present in the wheat extract. These pigments showed no adsorption onto the gel, thus regeneration is easily achieved, allowing its re-use. The enzyme also displayed strong salt-dependent adsorption onto adsorbents provided with pyridyIdisulfide moieties, but in this case enzyme binding was due to its thiol content since elution was achieved mainly through reduction with DTI'. When the enzyme was chromatographed on a series of hydrophobic alkyl ligands in the presence of 0. 5 M sodium sulphate, it was partially adsorbed on pentylagarose and quantitatively adsorbed on hexyt-agarose, elution being easily performed by sodium sulphate-free buffer. This behaviour was markedly different from that towards phenyl-Sepharose, to which the enzyme was strongly adsorbed and which required much more drastic elution conditions.

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Section: Resultssupporting

confidence: 84%