3 years ago

Molecular farming in barley: development of a novel production platform to produce human antimicrobial peptide LL-37

Alžbeta Mičúchová, Ivo Frébort, Edita Holásková, Marek Šebela, M. Tufan Öz, Petr Galuszka
The peptide LL-37, a component of the human innate immune system, represents a promising drug candidate. In particular, the development of low-cost production platform technology is a critical bottleneck in its use in medicine. In the present study, a viable approach for the LL-37 production in transgenic barley was developed. First, comparative analyses of the effects of different fused peptide epitope tags applicable for accumulation and purification on LL-37 production yield were performed using transient expression in tobacco leaves. Following the selection of the most yielding fusion peptide strategies, 8 different constructs for the expression of codon optimized chimeric LL-37 genes in transgenic barley plants were created. The expression of individual constructs was driven either by an endosperm-specific promoter of the barley B1 hordein gene or by the maize ubiquitin promoter. The transgenes were stably integrated into the barley genome and inherited in the subsequent generation. All transgenic lines showed normal phenotypes and were fertile. LL-37 accumulated in the barley seeds up to 0.55 mg per 1 kg of grain. The fused epitope tags were cleaved off by the use of enterokinase. Furthermore, in planta produced LL-37 including the fused versions was biologically active.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/biot.201700628

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