Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Protein Cyclization Enhanced Thermostability and Exopeptidase-Resistance of Green Fluorescent Protein

  • Zhao, Zhonglin (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences) ;
  • Ma, Xin (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences) ;
  • Li, Liang (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences) ;
  • Zhang, Wei (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences) ;
  • Ping, Shuzhen (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences) ;
  • Xu, Ming-Qun (New England Biolabs, Inc.) ;
  • Lin, Min (Biotechnology Research Institute, Chinese Academy of Agricultural Sciences)
  • Received : 2009.08.07
  • Accepted : 2009.10.04
  • Published : 2010.03.31
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Abstract

A mutant of green fluorescent protein ($GFPmut3^*$) from the jellyfish Aequorea victoria was cyclized in vitro and in vivo by the use of a naturally split intein from the dnaE gene of Synechocystis species PCC6803 (Ssp). Cyclization of $GFPmut3^*$ was confirmed by amino acid sequencing and resulted in an increased electrophoretic mobility compared with the linear $GFPmut3^*$. The circular $GFPmut3^*$ was $5^{\circ}C$ more thermostable than the linear form and significantly more resistant to proteolysis of exopeptidase. The circular $GFPmut3^*$ also displayed increased relative fluorescence intensity. In addition, chemical stability of $GFPmut3^*$ against GdnHCl revealed more stability of the circular form compared with the linear form.

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References

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