Some combinations of leucine zipper peptides are capable of forming a-helical heterodimeric coiled coils with very high affinity. These can be used as physical cross-linkers in the design of protein-based polymers that form supramolecular structures, for example hydrogels, upon mixing solutions containing the complementary blocks. Such two-component physical networks are of interest for many applications in biomedicine, pharmaceutics, and diagnostics. This article describes the efficient secretory production of A and B type leucine zipper peptides fused to protein-based polymers in Pichia pastoris. By adjusting the fermentation conditions, we were able to significantly reduce undesirable proteolytic degradation. The formation of A-B heterodimers in mixtures of the purified products was confirmed by size exclusion chromatography. Our results demonstrate that protein-based polymers incorporating functional heterodimer-forming blocks can be produced with P. pastoris in sufficient quantities for use in future supramolecular self-assembly studies and in various applications.
Domeradzka, N. E., Werten, M. W. T., de Vries, R. J., & de Wolf, F. A. (2016). Production in Pichia pastoris of protein-based polymers with small heterodimer-forming blocks. Biotechnology and Bioengineering, 113(5), 953-960. https://doi.org/10.1002/bit.25861