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Sequential Detection of Thermophilic Lipase and Protease by Zymography

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Zymography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1626))

Abstract

Lipase and protease present in cell-free fractions of thermophilic Bacillus sp. cultures were analyzed by polyacrylamide gel (PAG) electrophoresis. After run, the gel is electrotransferred to another PAG copolymerized with glycerol tributyrate, olive oil, and gelatin. This multi-substrate gel was incubated first for lipase detection, until bands appeared, and then stained with Coomassie for protease detection. Advantages of this sequential procedure are the detection of two different enzyme activities on a single PAG, beside time and resource saving.

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References

  1. Kwon MA, Kim HS, Hahm DH, Song JK (2011) Synthesis activity-based zymography for detection of lipases and esterases. Biotechnol Lett 33:741–746

    Article  CAS  PubMed  Google Scholar 

  2. Singh R, Gupta N, Goswami VK, Gupta R (2006) A simple activity staining protocol for lipases and esterases. Appl Microbiol Biotechnol 70:679–682

    Article  CAS  PubMed  Google Scholar 

  3. Wilkesman J, Kurz L (2009) Protease analysis by zymography: a review on techniques and patents. Recent Pat Biotechnol 3:175–184

    Article  CAS  PubMed  Google Scholar 

  4. Wilkesman J, Hernández Z, Fernández M, Contreras LM, Kurz L (2014) Enhancement of sequential zymography technique for the detection of thermophilic lipases and proteases. Amino Acids 46(5):1409–1413

    Article  CAS  PubMed  Google Scholar 

  5. Brahimi-Horn MC, Guglielmino ML, Gaal AM, Sparrow LG (1991) Nondenaturing protein electrotransfer of the esterase activity of lipolytic preparations. Anal Biochem 196:76–79

    Article  CAS  PubMed  Google Scholar 

  6. Pan D, Hill AP, Kashou A, Wilson KA, Tan-Wilson A (2011) Electrophoretic transfer protein Zymography. Anal Biochem 411:277–283

    Article  CAS  PubMed  Google Scholar 

  7. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  8. De Moreno MR, Smith JF, Smith RV (1985) Silver staining of proteins in polyacrylamide gels: increased sensitivity through a combined Coomassie blue-silver stain procedure. Anal Biochem 151:466–470

    Article  PubMed  Google Scholar 

  9. Kim MH, Kim HK, Lee JK, Park SY, Oh TK (2000) Thermostable lipase of Bacillus stearothermophilus: high-level production, purification, and calcium-dependent thermostability. Biosci Biotechnol Biochem 64:280–286

    Article  CAS  PubMed  Google Scholar 

  10. Manchenko GP (2003) Handbook of detection of enzymes on electrophorectic gels, 2nd edn. CRC Press LLC, Boca Raton, FL

    Google Scholar 

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Acknowledgments

This work was supported by the Consejo de Desarrollo Científico y Humanístico of the University of Carabobo (CDCH-UAC 373-2010) and was benefited by the Venezuelan Law of Science and Technology (LOCTI) and the Researcher Stimulation Program (PEII).

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Authors and Affiliations

  1. Center for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, Universidad de Carabobo, Valencia, Venezuela

    Liliana Kurz, Zully Hernández & Lellys M. Contreras

  2. Centre for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, Universidad de Carabobo, Valencia, Venezuela

    Jeff Wilkesman

Authors
  1. Liliana Kurz
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  2. Zully Hernández
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  3. Lellys M. Contreras
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  4. Jeff Wilkesman
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Corresponding author

Correspondence to Jeff Wilkesman .

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Editors and Affiliations

  1. Centre for Environmental, Biology and Chemistry Research, Facultad de Ciencias y Tecnología, University of Carabobo, Valencia, Venezuela

    Jeff Wilkesman

  2. Centre for Environmental, Biology and Chemistry Research Facultad de Ciencias y Tecnología, University of Carabobo, Valencia, Venezuela

    Liliana Kurz

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Kurz, L., Hernández, Z., Contreras, L.M., Wilkesman, J. (2017). Sequential Detection of Thermophilic Lipase and Protease by Zymography. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_25

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  • DOI: https://doi.org/10.1007/978-1-4939-7111-4_25

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7109-1

  • Online ISBN: 978-1-4939-7111-4

  • eBook Packages: Springer Protocols

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