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PubMed · 15007847

Recombinant protein purification from pea.

Todd J Menkhaus·Cynthia Pate·Anthony Krech·Charles E Glatz
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Abstract

To assess the suitability of transgenic peas as a host for protein production from the perspective of ease of recovery, a strain containing recombinant beta-glucuronidase with poly(histidine) tail (GUSH6) was evaluated for solubility of the target protein in relation to native components (proteins, carbohydrates, and phenolics). Recovery of the recombinant GUSH6 from aqueous extracts by immobilized metal affinity chromatography with coupled Co(2+) yielded a nearly pure product with IDA (enrichment factor (EF) = 260) or NTA (EF = 200) resin. Single-step recoveries were also possible by isoelectric precipitation (EF = 4), polyelectrolyte precipitation (EF = 1.5), and anion-exchange chromatography (EF = 3.1), but enrichment factors were low.

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Chemical FractionationGlucuronidaseHydrogen-Ion ConcentrationPisum sativumPlants, Genetically ModifiedProtein EngineeringRecombinant ProteinsSeedsComparative StudyEvaluation StudyJournal ArticleResearch Support, Non-U.S. Gov't

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Todd J Menkhaus, Cynthia Pate, Anthony Krech, Charles E Glatz. 2004-04-05. Recombinant protein purification from pea.. https://doi.org/10.1002/bit.20039

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