Premium
Wet‐milling transgenic maize seed for fraction enrichment of recombinant subunit vaccine
Author(s) -
Moeller Lorena,
TaylorVokes Raye,
Fox Steve,
Gan Qinglei,
Johnson Lawrence,
Wang Kan
Publication year - 2009
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.326
Subject(s) - steeping , endosperm , recombinant dna , food science , protein subunit , chemistry , escherichia coli , biology , biochemistry , chromatography , gene
The production of recombinant proteins in plants continues to be of great interest for prospective large‐scale manufacturing of industrial enzymes, nutrition products, and vaccines. This work describes fractionation by wet‐milling of transgenic maize expressing the B subunit of the heat‐labile enterotoxin of Escherichia coli (LT‐B), a potent immunogen and candidate for oral vaccine and vaccine components. The LT‐B gene was directed to express in seed by an endosperm specific promoter. Two steeping treatments, traditional steeping (TS, 0.2% SO 2 + 0.5% lactic acid) and water steeping (WS, water only), were evaluated to determine effects on recovery of functional LT‐B in wet‐milled fractions. The overall recovery of the LT‐B protein from WS treatment was 1.5‐fold greater than that from TS treatment. In both steeping types, LT‐B was distributed similarly among the fractions, resulting in enrichment of functional LT‐B in fine fiber, coarse fiber and pericarp fractions by concentration factors of 1.5 to 8 relative to the whole kernels on a per‐mass basis. Combined with endosperm‐specific expression and secretory pathway targeting, wet‐milling enables enrichment of high‐value recombinant proteins in low‐value fractions, such as the fine fiber, and co‐utilization of remaining fractions in alternative industrial applications. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010