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Purification, characterization, and N‐glycosylation of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures
Author(s) -
Corbin Jasmine M.,
Kailemia Muchena J.,
Cadieux C. Linn,
Alkanaimsh Salem,
Karuppanan Kalimuthu,
Rodriguez Raymond L.,
Lebrilla Carlito B.,
Cerasoli Douglas M.,
McDonald Karen A.,
Nandi Somen
Publication year - 2018
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26557
Subject(s) - recombinant dna , glycosylation , biochemistry , sialic acid , enzyme , chemistry , butyrylcholinesterase , glycan , xylose , downstream processing , chromatography , glycoprotein , fermentation , aché , acetylcholinesterase , gene
Recombinant butyrylcholinesterase produced in a metabolically regulated transgenic rice cell culture (rrBChE) was purified to produce a highly pure (95%), active form of enzyme. The developed downstream process uses common manufacturing friendly operations including tangential flow filtration, anion‐exchange chromatography, and affinity chromatography to obtain a process recovery of 42% active rrBChE. The purified rrBChE was then characterized to confirm its comparability to the native human form of the molecule (hBChE). The recombinant and native enzyme demonstrated comparable enzymatic behavior and had an identical amino acid sequence. However, rrBChE differs in that it contains plant‐type complex N‐glycans, including an α‐1,3 linked core fucose, and a β‐1,2 xylose, and lacking a terminal sialic acid. Despite this difference, rrBChE is demonstrated to be an effective stoichiometric bioscavenger for five different organophosphorous nerve agents in vitro. Together, the efficient downstream processing scheme and functionality of rrBChE confirm its promise as a cost‐effective alternative to hBChE for prophylactic and therapeutic use.

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