Effect of ion and surfactant choice on the recovery of a histidine-tagged protein from tobacco extract using foam fractionation

Czarena Crofcheck, Indu Maiti, Sitakanta Pattanaik, Michael Jay

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


Tobacco plants can be used for the production of proteins for pharmaceutical applications. One of the most difficult and expensive tasks associated with this technology is isolating the product of interest from the hundreds of other chemicals found in tobacco. We describe a new recovery strategy in which the protein of interest is "tagged" with a histidine structure, which forms a complex with metal ions and a surfactant that will accumulate in the foamate of a foam fractionation step. His-gus, a histidine-tagged enzyme, was selectively recovered in the presence of two different surfactants and two different metal ions. The foam fractionation with N-ε-dodecylamido-N-α, N-α,-bis(carboxymethyl)-L-lysine surfactant and Ni2+ ions resulted in an average His-gus activity recovery value of 88% and an activity enrichment of 2.27. The performance of the recovery strategy without tobacco extract resulted in an average activity recovery value of 63.32% and an average activity enrichment value of 5.16, utilizing lauroyl ethylenediaminetriacetate surfactant and Ni2+ ions. It was shown that even though a majority of the native tobacco proteins are removed during the prefoaming step, the presence of tobacco extract does affect the recovery of His-gus.

Original languageEnglish
Pages (from-to)79-91
Number of pages13
JournalApplied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology
Issue number1
StatePublished - Oct 2004

Bibliographical note

Funding Information:
We wish to thank Jeremie Wade and Courtney Fisk for their help in performing the experiments. The original paper (04-05-104) reports results of an investigation by the Kentucky Agricultural Experiment Station and is published with the approval of the director. We greatly appreciate the financial support provided by the Kentucky Science and Engineering Foundation (KSEF-148-502-02-07).


  • Downstream processing
  • Histidine-tagged protein
  • Protein recovery
  • Tobacco extract
  • Transgenic tobacco
  • Yeast extract

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology


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