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Application of fibrin zymography for determining the optimum culture time for protease activity

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Biotechnology Techniques

Abstract

The culture time for maximal enzyme production by bacteria was determined by SDS-fibrin zymography. Bacillus subtilis and B. amyloliquefaciens secreted several extracellular proteases during the cellular lysis after 40 h and 30 h, respectively. Using this technique, we determined which protein possessed activity and would be isolated before the process of purification began. This proposed method offers an effective harvesting time in which to obtain the proteins having maximal activity.

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References

  • Heussen C, Dowdle EB (1980) Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal. Biochem. 102: 196-202.

    Google Scholar 

  • Kim SH, Choi NS (1999) Electrophoretic analysis of protease inhibitors in fibrin zymography. Anal. Biochem. 270: 179-181.

    Google Scholar 

  • Kim WK, Choi KH, Kim YT, Park HH, Choi JY, Lee YS, Oh HI, Kwon IB, Lee SY (1996) Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from Chungkook-Jang. Appl. Environ. Microbiol. 62: 2482-2488.

    Google Scholar 

  • Kim SH, Choi NS, Lee WY (1998) Fibrin zymography: a direct analysis of fibrinolytic enzymes on gels. Anal. Biochem. 263: 115-116.

    Google Scholar 

  • Kleiner DE, Stetler-Stevenson WG (1994) Quantitative zymography: detection of picogram quantities of gelatinases. Anal. Biochem. 218: 325-329.

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680-685.

    Google Scholar 

  • Leber TM, Balkwill FR (1997) Zymography: a single-step staining method for quantitaiton of proteolytic activity on substrate gels. Anal. Biochem. 249: 24-28.

    Google Scholar 

  • Nakamura T, Yamagata Y, Ichishima E (1992) Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis (natto). Biosci. Biotechnol. Biochem. 56: 1869-1871.

    Google Scholar 

  • Raser KJ, Posner A, Wang KKW (1995) Casein zymography: a method to study u-Calpain, m-Calpain, and their inhibitory agents. Arch. Biochem. Biophys. 319: 211-216.

    Google Scholar 

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Authors and Affiliations

  1. Protein Function Research Unit, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon, 305-600, Korea

    Nack-Shick Choi & Seung-Ho Kim

Authors
  1. Nack-Shick Choi
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  2. Seung-Ho Kim
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Correspondence to Seung-Ho Kim.

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Choi, NS., Kim, SH. Application of fibrin zymography for determining the optimum culture time for protease activity. Biotechnology Techniques 13, 899–901 (1999). https://doi.org/10.1023/A:1008942611827

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  • DOI: https://doi.org/10.1023/A:1008942611827

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