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Facilitative production of an antimicrobial peptide royalisin and its antibody via an artificial oil‐body system
Author(s) -
Tseng JunMing,
Huang JunRu,
Huang HsiouChen,
Tzen Jason T. C.,
Chou WingMing,
Peng ChiChung
Publication year - 2010
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.528
Subject(s) - oleosin , intein , recombinant dna , fusion protein , escherichia coli , antimicrobial peptides , biochemistry , peptide , antibody , inclusion bodies , antimicrobial , cecropin , chemistry , biology , microbiology and biotechnology , rna , rna splicing , immunology , gene
Royalisin found in the royal jelly of Apis mellifera is an antimicrobial peptide (AMP). It has a molecular weight of 5.5 kDa, which contains six cysteine residues. In this study, royalisin was overexpressed in Escherichia coli AD494 (DE3) as two oleosin‐fusion proteins for preparation of its antibodies and functional purification. The recombinant royalisin, fused with oleosin central hydrophobic domain in both N‐ and C‐termini, was reconstituted with triacylglycerol and phospholipids to form artificial oil bodies (AOBs). The AOBs were then purified to raise the antibodies. These antibodies could recognize both the native and recombinant royalisins, but not oleosin. Another oleosin‐intein S ‐fusion protein was purified by AOBs system, and royalisin was subsequently released from the AOBs through self‐splicing of the intein. The recombinant royalisin exhibited high antibacterial activity, which suggested that it was refolded to its functional structure. These results demonstrated that AOBs system is an efficient method to functionally express and purify small AMPs. In addition, it also provides a facile platform for the production of antibodies against small peptides. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2011