Histidine tagged protein recovery from tobacco extract by foam fractionation.
Tobacco plants have the potential to be used for the production of proteins for pharmaceutical applications. This work describes a novel protein recovery strategy where the protein of interest is "tagged" with a histidine sequence, which forms a complex with cobalt ions and surfactant possessing a chelating functionality, such that the protein is recovered in the foamate of a foam fractionation step. His-gus, a histidine-tagged enzyme, was chosen as a model protein to study the feasibility of this strategy. The His-gus is recovered from spiked prefoamed tobacco extract by foam fractionation in the presence of surfactant and cobalt ions with an enrichment of 1.29 and a recovery of 21.5% in terms of an adjusted activity.
Crofcheck
Czarena
C
Department of Biosystems and Agricultural Engineering, College of Pharmacy, and Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, Kentucky 40546, USA. ccrofche@bae.uky.edu
Loiselle
Melanie
M
Weekley
James
J
Maiti
Indu
I
Pattanaik
Sitakanta
S
Bummer
Paul M
PM
Jay
Michael
M
D005591:Chemical Fractionation / Q000379:methods*; D004789:Enzyme Activation; D005240:Feasibility Studies; D005966:Glucuronidase / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D006639:Histidine / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013501:Surface-Active Agents / Q000737:chemistry*; D014026:Nicotiana / Q000737:chemistry* / Q000201:enzymology* / Q000235:genetics
D010936:Plant Extracts; D011993:Recombinant Fusion Proteins; D013501:Surface-Active Agents; D006639:Histidine; D005966:Glucuronidase