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KMID : 1094720190240040658
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Biotechnology and Bioprocess Engineering
2019 Volume.24 No. 4 p.658 ~ p.665
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Refolding with Simultaneous Purification of Recombinant Core Streptavidin Using Single-step High-performance Hydrophobic Interaction Chromatography
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Wang Siyao
Zhang Yuejuan
Gao Dong
Zi Jing
Wang Wenpeng
Zhang Nianzhe
Wan Yi
Wang Lili
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Abstract
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Streptavidin has applied to many areas including detection, purification, labeling, crosslinking and immobilization resulting in a high demand on its production. In this study, we report a method for preparation of recombinant core streptavidin (cSAV) protein using highperformance hydrophobic interaction chromatography (HPHIC). Firstly the cSAV was successfully cloned and expressed in Escherichia coli as inclusion bodies. A bifunctional stationary phase mainly working as HIC mode accompanied by weak anion exchange chromatography (WAX) was prepared using β-phenylethylamine (PEA) as a ligand. The denatured cSAV was then refolded and simultaneously purified by PEA hydrophobic interaction chromatography (PEA-HIC). The mass recovery and purity of cSAV by single-step were 30.2% and 98%, respectively. The bioactivity was determined to be 13.2 U/mg by biotin binding capacity assay. This method provides a new possibility for fast separation with simultaneous renaturation of cSAV.
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KEYWORD
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core streptavidin, high-performance hydrophobic interaction chromatography, β-phenylethylamine, inclusion body
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