title: Semicontinuous Bioreactor Production of Recombinant Butyrylcholinesterase in Transgenic Rice Cell Suspension Cultures.
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.
authors.affiliation: Chemical Engineering and Materials Science, University of California, Davis Davis, CA, USA.
authors.forename: Jasmine M
authors.initials: JM
authors.lastname: Corbin
authors.affiliation: Chemical Engineering and Materials Science, University of California, Davis Davis, CA, USA.
authors.forename: Bryce I
authors.initials: BI
authors.lastname: Hashimoto
authors.affiliation: Chemical Engineering and Materials Science, University of California, Davis Davis, CA, USA.
authors.forename: Kalimuthu
authors.initials: K
authors.lastname: Karuppanan
authors.affiliation: Chemical Engineering and Materials Science, University of California, Davis Davis, CA, USA.
authors.forename: Zachary R
authors.initials: ZR
authors.lastname: Kyser
authors.affiliation: Arcadia Biosciences Davis, CA, USA.
authors.forename: Liying
authors.initials: L
authors.lastname: Wu
authors.affiliation: Leidos, Inc. Frederick, MD, USA.
authors.forename: Brian A
authors.initials: BA
authors.lastname: Roberts
authors.affiliation: Leidos, Inc. Frederick, MD, USA.
authors.forename: Amy R
authors.initials: AR
authors.lastname: Noe
authors.affiliation: Global HealthShare®, Molecular and Cellular Biology, University of California, Davis Davis, CA, USA.
authors.forename: Raymond L
authors.initials: RL
authors.lastname: Rodriguez
authors.affiliation: Chemical Engineering and Materials Science, University of California, Davis Davis, CA, USA.
authors.forename: Karen A
authors.initials: KA
authors.lastname: McDonald
authors.affiliation: Global HealthShare®, Molecular and Cellular Biology, University of California, Davis Davis, CA, USA.
authors.forename: Somen
authors.initials: S
authors.lastname: Nandi
keywords: butyrylcholinesterase; inducible promoter; plant cell culture; rice amylase 3D; semicontinuous culture