Abstract

Esterases and lipases are lipolytic enzymes that catalyse the hydrolysis of triacylglycerols, Determination of lipolysis on agar plates is a simple approach to determine lipase or esterase action, but visual evaluation of lipolysis is frequently difficult in practice. Therefore, the aim of this work was to improve the efficiency of lipolysis visualization in tributyrin agar (mTBA) by adding calcium and/or magnesium ions in the screening of lipolytic microbial strains. Lipolytic activity was evaluated in mTBA using the well diffusion technique, where a clear zone around the inoculated wells indicated lipid hydrolysis. Results suggest that the addition of 2.5?mM calcium and 5.0?mM magnesium was the best combination of ion addition to TBA. Lipolytic activity increased the clearing zone up to 38% more than without the addition of ions and the clear zone was clearly observed. The mTBA plate was used with culture collection microbial strains, as well as with a collection of soil microorganisms, to identify lipase producers. The addition of calcium and magnesium ions can provide an easier screening procedure for selection of lipolytic bacterial strains.•A modified tributyrin agar for screening of lipolytic bacteria was prepared by adding calcium and magnesium ions.•The modified TBA agar was tested with control bacterial strains, and, based on the results, 2.5?mM Ca and 5.0?mM Mg ions were added in the mTBA.•mTBA was validated with environmental bacterial strains for screening of lipolytic activity.

Highlights

  • A modified tributyrin agar for screening of lipolytic bacteria was prepared by adding calcium and magnesium ions

  • For the purposes of the culture medium, we propose that most of the bacterial strains increased their lipolytic activity

  • After 48 h of incubation, strain RIA had a yellowish pigmentation over the halo. These pigmentations did not represent a problem for measuring lipolytic activity, but we consider that this observation can be important for further studies on the differential expression of pigments, since it might have a positive effect on lipolysis, including association with bio-surfactants [20,21,22]

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Summary

Method Article

Improved method for qualitative screening of lipolytic bacterial strains Jair Carrazco-Palafoxa,b, Blanca E. The aim of this work was to improve the efficiency of lipolysis visualization in tributyrin agar (mTBA) by adding calcium and/or magnesium ions in the screening of lipolytic microbial strains. Lipolytic activity was evaluated in mTBA using the well diffusion technique, where a clear zone around the inoculated wells indicated lipid hydrolysis. The addition of calcium and magnesium ions can provide an easier screening procedure for selection of lipolytic bacterial strains. A modified tributyrin agar for screening of lipolytic bacteria was prepared by adding calcium and magnesium ions. ARTICLE INFO Method name: Improved method for qualitative screening of lipolytic bacterial strains Keywords: Lipase activity, Calcium ion, Magnesium ion, Bioprospecting Article history: Received 5 September 2017; Accepted 18 January 2018; Available online 1 February 2018

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