Recombinant protein purification from pea.
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.
Department of Chemical Engineering, Iowa State University, Ames, Iowa 50011-2230, USA.
Todd J
TJ
Menkhaus
Cynthia
C
Pate
Anthony
A
Krech
Charles E
CE
Glatz
D005591:Chemical Fractionation / Q000379:methods; D005966:Glucuronidase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D018532:Pisum sativum / Q000201:enzymology* / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D012639:Seeds / Q000201:enzymology / Q000235:genetics
D011994:Recombinant Proteins; D005966:Glucuronidase