Details

Autor(en) / Beteiligte

• Tian, Ernuo
• Shen, Xiao
• Xiao, Meili
• Zhu, Zhihua
• Yang, Yi
• Yan, Xing
• Wang, Pingping
• Zou, Gen
• Zhou, Zhihua

Titel

An engineered Pichia pastoris platform for the biosynthesis of silk-based nanomaterials with therapeutic potential

Ist Teil von

• International journal of biological macromolecules, 2024-06, Vol.269 (Pt 2), p.131954-131954, Article 131954

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2024

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Silk fibroin (SF) from the cocoon of silkworm has exceptional mechanical properties and biocompatibility and is used as a biomaterial in a variety of fields. Sustainable, affordable, and scalable manufacturing of SF would enable its large-scale use. We report for the first time the high-level secretory production of recombinant SF peptides in engineered Pichia pastoris cell factories and the processing thereof to nanomaterials. Two SF peptides (BmSPR3 and BmSPR4) were synthesized and secreted by P. pastoris using signal peptides and appropriate spacing between hydrophilic sequences. By strain engineering to reduce protein degradation, increase glycyl-tRNA supply, and improve protein secretion, we created the optimized P. pastoris chassis PPGSP-8 to produce BmSPR3 and BmSPR4. The SF fed-batch fermentation titers of the resulting two P. pastoris cell factories were 11.39 and 9.48 g/L, respectively. Protein self-assembly was inhibited by adding Tween 80 to the medium. Recombinant SF peptides were processed to nanoparticles (NPs) and nanofibrils. The physicochemical properties of nanoparticles R3NPs and R4NPs from the recombinant SFs synthesized in P. pastoris cell factories were similar or superior to those of RSFNPs (Regenerated Silk Fibroin NanoParticles) originating from commercially available SF. Our work will facilitate the production by microbial fermentation of functional SF for use as a biomaterial. •Pichia pastoris was engineered for high-level silk protein production and secretion.•More than 10 g/L silk fibroin protein can be produced by engineered cell factory.•Self-aggregation of silk fibroin proteins was inhibited by Tween 80.•Nanomaterials produced by recombinant silk fibroin protein exhibited similar properties as regenerative silk fibroin nanomaterials.•Recombinant silk fibroin nanoparticles have excellent drug delivery efficiency.