High-yield production of a super-soluble miniature spidroin for biomimetic high-performance materials

MATERIALS TODAY

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Article

Schmuck, Benjamin¹  Greco, Gabriele^2,3  Barth, Andreas⁴  Pugno, Nicola M.^2,5  Johansson, Jan¹  Rising, Anna^1,3 

[1] Karolinska Inst, Dept Biosci & Nutr, S-14186 Huddinge, Sweden

[2] Univ Trento, Dept Civil Environm & Mech Engn, Lab Bioinspired Bion Nano Meta Mat & Mech, Via Mesiano 77, I-38123 Trento, Italy

[3] Swedish Univ Agr Sci, Dept Anat Physiol & Biochem, Uppsala, Sweden

[4] Stockholm Univ, Dept Biochem & Biophys, Svante Arrhenius Vag 16C, S-10691 Stockholm, Sweden

[5] Queen Mary Univ London, Sch Engn & Mat Sci, Mile End Rd, London E1 4N5, England

关键词: Artificial spider silk;    Spidroins;    Bioreactor cultivation;    Recombinant protein expression;   

DOI  :  10.1016/j.mattod.2021.07.020

来源: Elsevier

【 摘 要 】

The mechanical properties of artificial spider silks are approaching a stage where commercial applications become realistic. However, the yields of recombinant silk proteins that can be used to produce fibers with good mechanical properties are typically very low and many purification and spinning protocols still require the use of urea, hexafluoroisopropanol, and/or methanol. Thus, improved production and spinning methods with a minimal environmental impact are needed. We have previously developed a miniature spider silk protein that is characterized by high solubility in aqueous buffers and spinnability in biomimetic set-ups. In this study, we developed a production protocol that resulted in an expression level of >20 g target protein per liter in an Escherichia coli fedbatch culture, and subsequent purification under native conditions yielded 14.5 g/l. This corresponds to a nearly six-fold increase in expression levels, and a 10-fold increase in yield after purification compared to reports for recombinant spider silk proteins. Biomimetic spinning using only aqueous buffers resulted in fibers with a toughness modulus of 74 MJ/m(3), which is the highest reported for biomimetically as-spun artificial silk fibers. Thus, the process described herein represents a milestone for the economic production of biomimetic silk fibers for industrial applications.

【 授权许可】

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