title: Production in Pichia pastoris of protein-based polymers with small heterodimer-forming blocks.
abstract: Some combinations of leucine zipper peptides are capable of forming α-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.
authors.affiliation: Wageningen UR Food & Biobased Research, Bornse Weilanden 9, 6708 WG, Wageningen, The Netherlands.|Physical Chemistry and Soft Matter, Wageningen University and Research Centre, Dreijenplein 6, 6703 HB, Wageningen, The Netherlands.
authors.forename: Natalia E
authors.initials: NE
authors.lastname: Domeradzka
authors.affiliation: Wageningen UR Food & Biobased Research, Bornse Weilanden 9, 6708 WG, Wageningen, The Netherlands. marc.werten@wur.nl.
authors.forename: Marc W T
authors.initials: MW
authors.lastname: Werten
authors.affiliation: Physical Chemistry and Soft Matter, Wageningen University and Research Centre, Dreijenplein 6, 6703 HB, Wageningen, The Netherlands.
authors.forename: Renko
authors.initials: R
authors.lastname: de Vries
authors.affiliation: Wageningen UR Food & Biobased Research, Bornse Weilanden 9, 6708 WG, Wageningen, The Netherlands.
authors.forename: Frits A
authors.initials: FA
authors.lastname: de Wolf
mesh_terms: D000595:Amino Acid Sequence; D005285:Fermentation; D007218:Industrial Microbiology; D016350:Leucine Zippers; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*; D015202:Protein Engineering; D055503:Protein Multimerization; D059748:Proteolysis; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D010455:Peptides; D011108:Polymers; D011993:Recombinant Fusion Proteins
keywords: Pichia pastoris; molecular self-assembly; protein secretion; protein-based polymers; proteolysis