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Detection of Multiple Enzymes in Fermentation Broth Using Single PAGE Analysis

  • K. Divakar
  • J. Deepa Arul Priya
  • G. Panneer Selvam
  • M. Suryia Prabha
  • Ashwin Kannan
  • G. Nandhini Devi
  • Pennathur Gautam
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1853)

Abstract

Activity staining or zymography is a technique to detect enzymes based on their function/activity toward a specific substrate. Multiple enzyme-producing microbes secrete enzymes along with other proteins at varying time points during fermentation. The technique of zymography can be used to detect functionality of enzymes in complex protein/other enzyme mixtures. The protein bands corresponding to specific enzyme among other enzymes/proteins can be located by polyacrylamide gel electrophoresis (PAGE) followed by zymogram analysis. This can be employed to locate the secretion pattern of protein/enzyme from intracellular region to extracellular medium. Here we describe simple method for detection and cellular localization of esterases and protease secreted by single microbial strain in one PAGE gel.

Key words

Zymogram Multiple enzymes Fermentation broth PAGE 

Notes

Acknowledgments

We thank Department of Biotechnology and Department of Science and Technology (DST), Government of India, for their continuous support to carry out research in our lab. K. Divakar thanks DST, Govt. of India for funding through DST-INSPIRE Faculty award and research grant (IFA14-ENG-87).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • K. Divakar
    • 1
  • J. Deepa Arul Priya
    • 2
  • G. Panneer Selvam
    • 2
  • M. Suryia Prabha
    • 2
  • Ashwin Kannan
    • 3
  • G. Nandhini Devi
    • 2
  • Pennathur Gautam
    • 2
    • 4
  1. 1.Department of BiotechnologyNational Institute of TechnologyWarangalIndia
  2. 2.Centre for BiotechnologyAnna UniversityChennaiIndia
  3. 3.Department of Chemical EngineeringVisvesvaraya National Institute of TechnologyNagpurIndia
  4. 4.AU-KBC Research CentreAnna UniversityChennaiIndia

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