Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Identification of Recombinant Subtilisins

  • CHOI , NACK-SHICK (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology, Department of Microbiology, Chungnam National University) ;
  • YOO, KI-HYUN (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • YOON, KAB-SEOG (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • CHANG, KYU-TAE (Primate Research Laboratory, Korea Research Institute of Bioscience and Biotechnology) ;
  • MAENG, PIL-JAE (Department of Microbiology, Chungnam National University) ;
  • KIM, SEUNG-HO (Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology)
  • Published : 2005.02.01
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Abstract

To identify the activity of recombinant subtilisins (subtilisin BPN' and subtilisin Carlsberg), three different zymography methods, SDS-fibrin zymography (SDS-FZ), reverse fibrin zymography (RFZ), and isoelectric focusingfibrin zymography (IEF-FZ), were used. The recombinant subtilisins BPN' and Carlsberg did not migrate into the electrophoretic field based on a Laemmli buffer system, instead forming a "binding mode" at the top part of the separating gels with the SDS-FZ and RFZ techniques. Yet, this problem was resolved when using IEF-FZ with a pH range from 3 to 10. In addition, all these methods enabled the activity of a recombinant pro-subtilisin DJ-4 to be detected without a refolding pathway.

Keywords

References

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