Engineered a novel pH-sensitive short major ampullate spidroin

Chen Zhang; Jiali Mi; Haishan Qi; Jie Huang; Simin Liu; Lei Zhang; Daidi Fan

doi:10.1016/j.ijbiomac.2020.03.153

Engineered a novel pH-sensitive short major ampullate spidroin

Int J Biol Macromol. 2020 Jul 1:154:698-705. doi: 10.1016/j.ijbiomac.2020.03.153. Epub 2020 Mar 18.

Authors

Chen Zhang¹, Jiali Mi¹, Haishan Qi², Jie Huang¹, Simin Liu¹, Lei Zhang³, Daidi Fan⁴

Affiliations

¹ Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin 300072, China.
² Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin 300072, China. Electronic address: hsqi@tju.edu.cn.
³ Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin 300072, China; Qingdao Institute for Marine Technology of Tianjin University, Qingdao 266235, China. Electronic address: lei_zhang@tju.edu.cn.
⁴ Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Taibai North Road 229, Xi'an, Shaanxi 710069, China.

PMID: 32198037
DOI: 10.1016/j.ijbiomac.2020.03.153

Abstract

The pH diversification has been proved as an important factor affecting the self-assembly of spidroin. Herein, we constructed a novel spider silk protein (NT-MaSp1s-CT) with the pH-dependent secondary structures, containing pH-sensitive N-terminal, C-terminal domains and a repeated core region with merely 191 amino acids. Then pH sensitivity of NT-MaSp1s-CT was detected at different pH conditions and NT-MaSp1s-CT displayed pH-dependent conformational transitions consistent with rational designed objective. Besides, the micelles theory was employed to inquiry the assembly mechanism of NT-MaSp1s-CT in high concentration spinning dope. As expected, NT-MaSp1s-CT protein can be spun into continuous and uniform fibers with the pH ranging from 2 to 11, which is the largest pH boundary for artificial spider silk formation, simplifying the assembly conditions and paving a broad path for spinning process. Moreover, the hemolysis and cytotoxicity of NT-MaSp1s-CT fibers were also determined and the novel fibers exhibit excellent biocompatibility, providing wider potential applications in the biomedical and pharmaceutical fields.

Keywords: Biocompatibility; Micelle; Recombinant spider silk protein; Synthetic biology; pH-sensitive.

MeSH terms

Animals
Fibroins / chemistry*
Hydrogen-Ion Concentration
Protein Engineering*
Protein Structure, Secondary
Recombinant Proteins / chemistry*
Spiders

Substances

Recombinant Proteins
Fibroins