Zymography Detection of a Bacterial Extracellular Thermoalkaline Esterase/Lipase Activity

Methods Mol Biol. 2017:1626:295-300. doi: 10.1007/978-1-4939-7111-4_28.

Abstract

Lipases are esterases that occur widely in nature, yet those with commercial relevance are exclusively from microbial origin. Glycerol and long-chain fatty acids are the products after hydrolysis of esters bonds in saponifiable lipids catalyzed by lipases. In this work, we describe lipase/esterase activity contained in cell-free fractions from thermophilic bacteria, cultured in medium containing olive oil. Analysis of the cell-free fractions by electrotransference zymography, using tributyrin as substrate, revealed bands corresponding to lipase activity. The method is simple, fast, and inexpensive.

Keywords: Electrophoresis; Electrotransference; Esterase; Lipases; Thermophiles; Zymography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Enzyme Assays / methods*
  • Esterases / metabolism*
  • Geobacillus / enzymology*
  • Geobacillus / metabolism
  • Hydrolysis
  • Lipase / metabolism*
  • Substrate Specificity
  • Triglycerides / metabolism

Substances

  • Triglycerides
  • Esterases
  • Lipase
  • tributyrin