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

13 Citations (Scopus)

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

Fingerprint

cholinesterase
bioreactors
cell suspension culture
genetically modified organisms
rice
callus
Western blotting
cells
screening
assays
enzymes

Keywords

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

ASJC Scopus subject areas

  • Plant Science

Cite this

Corbin, J. M., Hashimoto, B. I., Karuppanan, K., Kyser, Z. R., Wu, L., Roberts, B. 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

Semicontinuous bioreactor production of recombinant butyrylcholinesterase in transgenic rice cell suspension cultures. / Corbin, Jasmine M.; Hashimoto, Bryce I.; Karuppanan, Kalimuthu; Kyser, Zachary R.; Wu, Liying; Roberts, Brian A.; Noe, Amy R.; Rodriguez, Raymond L.; McDonald, Karen A.; Nandi, Somen.

In: Frontiers in Plant Science, Vol. 7, No. MAR2016, 412, 31.03.2016.

Research output: Contribution to journalArticle

Corbin, JM, Hashimoto, BI, Karuppanan, K, Kyser, ZR, Wu, L, Roberts, BA, Noe, AR, Rodriguez, RL, McDonald, KA & Nandi, S 2016, 'Semicontinuous bioreactor production of recombinant butyrylcholinesterase in transgenic rice cell suspension cultures', Frontiers in Plant Science, vol. 7, no. MAR2016, 412. https://doi.org/10.3389/fpls.2016.00412
Corbin, Jasmine M. ; Hashimoto, Bryce I. ; Karuppanan, Kalimuthu ; Kyser, Zachary R. ; Wu, Liying ; Roberts, Brian A. ; Noe, Amy R. ; Rodriguez, Raymond L. ; McDonald, Karen A. ; Nandi, Somen. / Semicontinuous bioreactor production of recombinant butyrylcholinesterase in transgenic rice cell suspension cultures. In: Frontiers in Plant Science. 2016 ; Vol. 7, No. MAR2016.
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