Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates

Abstract Lipases are currently used in food technology for the modification of fats and oils. The thermal stability of lipase is an essential characteristic for this application. This study compares four individual single-step methods (heat treatment, ethanol precipitation, ammonium sulfate precipit...

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Main Authors: Marie Kathleen Walsh, Teif AbdulQani Najm
Format: Article
Language:English
Published: Instituto de Tecnologia do Paraná (Tecpar) 2021-11-01
Series:Brazilian Archives of Biology and Technology
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132021000100221&tlng=en
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author Marie Kathleen Walsh
Teif AbdulQani Najm
author_facet Marie Kathleen Walsh
Teif AbdulQani Najm
author_sort Marie Kathleen Walsh
collection DOAJ
description Abstract Lipases are currently used in food technology for the modification of fats and oils. The thermal stability of lipase is an essential characteristic for this application. This study compares four individual single-step methods (heat treatment, ethanol precipitation, ammonium sulfate precipitation, and size-exclusion chromatography) to enrich lipase concentrations from thermophilic bacterial (Geobacillius stearothermophilus and Anoxybacillus flavithermus) cell lysates. SDS-PAGE and size exclusion chromatography were used to determine the molecular weights of the lipases and the enrichment efficiencies were determined using specific enzyme activities. The molecular weight of G. stearothermophilus lipase was approximately 42 kDa, and approximately 33 kDa for A. flavithermus lipase. For each organism, ethanol precipitation resulted in the highest enrichment fold, followed by ammonium sulfate precipitation, gel filtration and heat treatment respectively. The highest yields for G. stearothermophilus lipase were obtained with ammonium sulfate precipitation, followed by get filtration, and ethanol precipitation respectively. The highest yields for A. flavithermus lipase were obtained from heat treatment followed by ammonium sulfate precipitation, gel filtration and ethanol precipitation respectively. Ethanol precipitation and heat treatment are simple methods for enzyme enrichment from cell lysates and can result in high enzyme yields with moderate enrichment folds compared to complex multi-step purification methods.
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spelling doaj.art-267cb68e9cef485c90e82ac69887483f2022-12-21T19:35:59ZengInstituto de Tecnologia do Paraná (Tecpar)Brazilian Archives of Biology and Technology1678-43242021-11-016410.1590/1678-4324-2021200045Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell LysatesMarie Kathleen Walshhttps://orcid.org/0000-0002-7323-5930Teif AbdulQani Najmhttps://orcid.org/0000-0002-8040-1335Abstract Lipases are currently used in food technology for the modification of fats and oils. The thermal stability of lipase is an essential characteristic for this application. This study compares four individual single-step methods (heat treatment, ethanol precipitation, ammonium sulfate precipitation, and size-exclusion chromatography) to enrich lipase concentrations from thermophilic bacterial (Geobacillius stearothermophilus and Anoxybacillus flavithermus) cell lysates. SDS-PAGE and size exclusion chromatography were used to determine the molecular weights of the lipases and the enrichment efficiencies were determined using specific enzyme activities. The molecular weight of G. stearothermophilus lipase was approximately 42 kDa, and approximately 33 kDa for A. flavithermus lipase. For each organism, ethanol precipitation resulted in the highest enrichment fold, followed by ammonium sulfate precipitation, gel filtration and heat treatment respectively. The highest yields for G. stearothermophilus lipase were obtained with ammonium sulfate precipitation, followed by get filtration, and ethanol precipitation respectively. The highest yields for A. flavithermus lipase were obtained from heat treatment followed by ammonium sulfate precipitation, gel filtration and ethanol precipitation respectively. Ethanol precipitation and heat treatment are simple methods for enzyme enrichment from cell lysates and can result in high enzyme yields with moderate enrichment folds compared to complex multi-step purification methods.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132021000100221&tlng=enthermophilic lipaseenzyme enrichmentzymography
spellingShingle Marie Kathleen Walsh
Teif AbdulQani Najm
Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates
Brazilian Archives of Biology and Technology
thermophilic lipase
enzyme enrichment
zymography
title Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates
title_full Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates
title_fullStr Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates
title_full_unstemmed Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates
title_short Comparison of Single-Step Methods to Enrich Lipase Concentrations in Bacterial Cell Lysates
title_sort comparison of single step methods to enrich lipase concentrations in bacterial cell lysates
topic thermophilic lipase
enzyme enrichment
zymography
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132021000100221&tlng=en
work_keys_str_mv AT mariekathleenwalsh comparisonofsinglestepmethodstoenrichlipaseconcentrationsinbacterialcelllysates
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