Direct recovery of enhanced green fluorescent protein from unclarified Escherichia coli homogenate using ion exchange chromatography in stirred fluidized bed

Cher Pin Song, Chien Wei Ooi, Beng Ti Tey, Chong Xuan Lu, Bing Lan Liu, Yu Kaung Chang

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

A stirred fluidized bed (SFB) ion exchange chromatography was successfully applied in the direct recovery of recombinant enhanced green fluorescent protein (EGFP) from the unclarified Escherichia coli homogenate. Optimal conditions for both adsorption and elution processes were determined from the packed-bed adsorption systems conducted at a small scale using the clarified cell homogenate. The maximal adsorption capacity and dissociation constant for EGFP-adsorbent complex were found to be 6.3 mg/mL and 1.3 × 10−3 mg/mL, respectively. In an optimal elution of EGFP with 0.2 M of NaCl solution (pH 9) and at 200 cm/h, the recovery percent of the EGFP was approximately 93%. The performances of SFB chromatography for direct recovery of EGFP was also evaluated under different loading volumes (50–200 mL) of crude cell homogenate. The single-step purification of EGFP by SFB recorded in a high yield (95–98%) and a satisfactory purification factor (~3 folds) of EGFP from the cell homogenate at 200 rpm of rotating speed.

Original languageEnglish
Pages (from-to)4455-4465
Number of pages11
JournalInternational Journal of Biological Macromolecules
Volume164
DOIs
Publication statusPublished - 1 Dec 2020

Keywords

  • Direct recovery
  • Enhanced green fluorescent protein
  • Escherichia coli homogenate
  • Stirred fluidized bed adsorption
  • STREAMLINE DEAE adsorbent

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