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

文献类型： 外文期刊

作者： Tian, Ernuo 1 ; Shen, Xiao 2 ; Xiao, Meili 2 ; Zhu, Zhihua 2 ; Yang, Yi 1 ; Yan, Xing 2 ; Wang, Pingping 2 ; Zou, Gen 2 ; Zhou, Zhihua 1 ;

作者机构： 1.East China Univ Sci & Technol, Sch Pharm, Shanghai 200037, Peoples R China

2.Chinese Acad Sci, CAS Ctr Excellence Mol Plant Sci, CAS Key Lab Synthet Biol, Shanghai 200032, Peoples R China

3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China

4.Shanghai Acad Agr Sci, Inst Edible Fungi, Shanghai Key Lab Agr Genet & Breeding, Shanghai 201403, Peoples R China

关键词： Pichia pastoris; Silk fibroin; Secretion; Nanomaterials; Nanoparticles; Nanofibrils

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.2； 五年影响因子：7.8 ）

ISSN： 0141-8130

年卷期： 2024 年 269 卷

页码：

收录情况： SCI

摘要： 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 glycyltRNA 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.

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