Semicontinuous bioreactor production of recombinant butyrylcholinesterase in transgenic rice cell suspension cultures

Jasmine M. Corbin, Bryce I. Hashimoto, Kalimuthu Karuppanan, Zachary R. Kyser, Liying Wu, Brian A. Roberts, Amy R. Noe, Raymond L. Rodriguez, Karen A. McDonald, Somen Nandi

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

An active and tetrameric form of recombinant butyrylcholinesterase (BChE), a large and complex human enzyme, was produced via semicontinuous operation in a transgenic rice cell suspension culture. After transformation of rice callus and screening of transformants, the cultures were scaled up from culture flask to a lab scale bioreactor. The bioreactor was operated through two phases each of growth and expression. The cells were able to produce BChE during both expression phases, with a maximum yield of 1.6 mg BChE/L of culture during the second expression phase. Cells successfully regrew during a 5-day growth phase. A combination of activity assays and Western blot analysis indicated production of an active and fully assembled tetramer of BChE.

Original languageEnglish (US)
Article number412
JournalFrontiers in Plant Science
Volume7
Issue numberMAR2016
DOIs
StatePublished - Mar 31 2016

Keywords

  • Butyrylcholinesterase
  • Inducible promoter
  • Plant cell culture
  • Rice amylase 3D
  • Semicontinuous culture

ASJC Scopus subject areas

  • Plant Science

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    Corbin, J. M., Hashimoto, B. I., Karuppanan, K., Kyser, Z. R., Wu, L., Roberts, B. A., Noe, A. R., Rodriguez, R. L., McDonald, K. A., & Nandi, S. (2016). Semicontinuous bioreactor production of recombinant butyrylcholinesterase in transgenic rice cell suspension cultures. Frontiers in Plant Science, 7(MAR2016), [412]. https://doi.org/10.3389/fpls.2016.00412