KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Simple Purification of BA-RGD Protein Based on CaCl2/EDTA Treatment and Inclusion Body Washing

CaCl2/EDTA 및 비이온성 계면활성제 활용 Inclusion Body 정제법을 이용한 BA-RGD 단백질의 생산

  • Song, Wooho (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) ;
  • Byun, Chang Woo (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) ;
  • Yoon, Minho (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) ;
  • Eom, Ji Hoon (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) ;
  • Choi, Yoo Seong (Department of Chemical Engineering and Applied Chemistry, Chungnam National University)
  • 송우호 (충남대학교 응용화학공학과) ;
  • 변창우 (충남대학교 응용화학공학과) ;
  • 윤민호 (충남대학교 응용화학공학과) ;
  • 엄지훈 (충남대학교 응용화학공학과) ;
  • 최유성 (충남대학교 응용화학공학과)
  • Received : 2015.10.01
  • Accepted : 2015.11.07
  • Published : 2015.12.27
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Abstract

The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

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References

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