A spectrophotometric transesterification-based assay for lipases in organic solvent
A new method to evaluate lipase activities in nonaqueous conditions using vinyl ester absorbance at ultraviolet (UV) wavelengths is described. The model reaction is the transesterification between vinyl stearate and pentanol in hexane at 30 _C or in decane at 50 _C. The conversion of vinyl stearate into pentyl stearate is monitored through decreasing UV absorbance at 200 nm. Six commercial lipases were tested with this method, and results were compared with gas chromatography (GC) quantification and a classical spectrophotometric method using p-nitrophenyl palmitate. Results from the new spectrophotometric assay are similar both to results from GC quantification (R2 = 0.999) and to results from p-nitrophenyl palmitate (R2 = 0.989). The proposed method is able to evaluate both high activity from immobilized lipases such as immobilized Candida antarctica B lipase (3060 ± 350 U g_1) and low activity from crude enzymatic extracts such as Carica papaya dried latex (0.1 ± 0.04 U g_1). The method has also been used to measure kinetic parameters of C. antarctica B lipase for vinyl stearate and the correlation between its synthesis activity and its concentration. The method has also proved to be effective in studying the acyl selectivity of a lipase by comparing its activities with increasing chain lengths of vinyl esters.
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Subjects: | Q02 - Traitement et conservation des produits alimentaires, 000 - Autres thèmes, estérase, spectrométrie, latex, solvant, lipase, http://aims.fao.org/aos/agrovoc/c_2670, http://aims.fao.org/aos/agrovoc/c_7283, http://aims.fao.org/aos/agrovoc/c_4214, http://aims.fao.org/aos/agrovoc/c_7236, http://aims.fao.org/aos/agrovoc/c_12194, |
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dig-cirad-fr-5472762024-01-28T16:34:51Z http://agritrop.cirad.fr/547276/ http://agritrop.cirad.fr/547276/ A spectrophotometric transesterification-based assay for lipases in organic solvent. Goujard Laurent, Villeneuve Pierre, Baréa Bruno, Lecomte Jérôme, Pina Michel, Claude Sylvain, Le Petit Jean, Ferré Elisée. 2009. Analytical Biochemistry, 385 (1) : 161-167.https://doi.org/10.1016/j.ab.2008.10.025 <https://doi.org/10.1016/j.ab.2008.10.025> A spectrophotometric transesterification-based assay for lipases in organic solvent Goujard, Laurent Villeneuve, Pierre Baréa, Bruno Lecomte, Jérôme Pina, Michel Claude, Sylvain Le Petit, Jean Ferré, Elisée eng 2009 Analytical Biochemistry Q02 - Traitement et conservation des produits alimentaires 000 - Autres thèmes estérase spectrométrie latex solvant lipase http://aims.fao.org/aos/agrovoc/c_2670 http://aims.fao.org/aos/agrovoc/c_7283 http://aims.fao.org/aos/agrovoc/c_4214 http://aims.fao.org/aos/agrovoc/c_7236 http://aims.fao.org/aos/agrovoc/c_12194 A new method to evaluate lipase activities in nonaqueous conditions using vinyl ester absorbance at ultraviolet (UV) wavelengths is described. The model reaction is the transesterification between vinyl stearate and pentanol in hexane at 30 _C or in decane at 50 _C. The conversion of vinyl stearate into pentyl stearate is monitored through decreasing UV absorbance at 200 nm. Six commercial lipases were tested with this method, and results were compared with gas chromatography (GC) quantification and a classical spectrophotometric method using p-nitrophenyl palmitate. Results from the new spectrophotometric assay are similar both to results from GC quantification (R2 = 0.999) and to results from p-nitrophenyl palmitate (R2 = 0.989). The proposed method is able to evaluate both high activity from immobilized lipases such as immobilized Candida antarctica B lipase (3060 ± 350 U g_1) and low activity from crude enzymatic extracts such as Carica papaya dried latex (0.1 ± 0.04 U g_1). The method has also been used to measure kinetic parameters of C. antarctica B lipase for vinyl stearate and the correlation between its synthesis activity and its concentration. The method has also proved to be effective in studying the acyl selectivity of a lipase by comparing its activities with increasing chain lengths of vinyl esters. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/547276/1/document_547276.pdf application/pdf Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1016/j.ab.2008.10.025 10.1016/j.ab.2008.10.025 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ab.2008.10.025 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1016/j.ab.2008.10.025 |
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Q02 - Traitement et conservation des produits alimentaires 000 - Autres thèmes estérase spectrométrie latex solvant lipase http://aims.fao.org/aos/agrovoc/c_2670 http://aims.fao.org/aos/agrovoc/c_7283 http://aims.fao.org/aos/agrovoc/c_4214 http://aims.fao.org/aos/agrovoc/c_7236 http://aims.fao.org/aos/agrovoc/c_12194 Q02 - Traitement et conservation des produits alimentaires 000 - Autres thèmes estérase spectrométrie latex solvant lipase http://aims.fao.org/aos/agrovoc/c_2670 http://aims.fao.org/aos/agrovoc/c_7283 http://aims.fao.org/aos/agrovoc/c_4214 http://aims.fao.org/aos/agrovoc/c_7236 http://aims.fao.org/aos/agrovoc/c_12194 |
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Q02 - Traitement et conservation des produits alimentaires 000 - Autres thèmes estérase spectrométrie latex solvant lipase http://aims.fao.org/aos/agrovoc/c_2670 http://aims.fao.org/aos/agrovoc/c_7283 http://aims.fao.org/aos/agrovoc/c_4214 http://aims.fao.org/aos/agrovoc/c_7236 http://aims.fao.org/aos/agrovoc/c_12194 Q02 - Traitement et conservation des produits alimentaires 000 - Autres thèmes estérase spectrométrie latex solvant lipase http://aims.fao.org/aos/agrovoc/c_2670 http://aims.fao.org/aos/agrovoc/c_7283 http://aims.fao.org/aos/agrovoc/c_4214 http://aims.fao.org/aos/agrovoc/c_7236 http://aims.fao.org/aos/agrovoc/c_12194 Goujard, Laurent Villeneuve, Pierre Baréa, Bruno Lecomte, Jérôme Pina, Michel Claude, Sylvain Le Petit, Jean Ferré, Elisée A spectrophotometric transesterification-based assay for lipases in organic solvent |
description |
A new method to evaluate lipase activities in nonaqueous conditions using vinyl ester absorbance at ultraviolet (UV) wavelengths is described. The model reaction is the transesterification between vinyl stearate and pentanol in hexane at 30 _C or in decane at 50 _C. The conversion of vinyl stearate into pentyl stearate is monitored through decreasing UV absorbance at 200 nm. Six commercial lipases were tested with this method, and results were compared with gas chromatography (GC) quantification and a classical spectrophotometric method using p-nitrophenyl palmitate. Results from the new spectrophotometric assay are similar both to results from GC quantification (R2 = 0.999) and to results from p-nitrophenyl palmitate (R2 = 0.989). The proposed method is able to evaluate both high activity from immobilized lipases such as immobilized Candida antarctica B lipase (3060 ± 350 U g_1) and low activity from crude enzymatic extracts such as Carica papaya dried latex (0.1 ± 0.04 U g_1). The method has also been used to measure kinetic parameters of C. antarctica B lipase for vinyl stearate and the correlation between its synthesis activity and its concentration. The method has also proved to be effective in studying the acyl selectivity of a lipase by comparing its activities with increasing chain lengths of vinyl esters. |
format |
article |
topic_facet |
Q02 - Traitement et conservation des produits alimentaires 000 - Autres thèmes estérase spectrométrie latex solvant lipase http://aims.fao.org/aos/agrovoc/c_2670 http://aims.fao.org/aos/agrovoc/c_7283 http://aims.fao.org/aos/agrovoc/c_4214 http://aims.fao.org/aos/agrovoc/c_7236 http://aims.fao.org/aos/agrovoc/c_12194 |
author |
Goujard, Laurent Villeneuve, Pierre Baréa, Bruno Lecomte, Jérôme Pina, Michel Claude, Sylvain Le Petit, Jean Ferré, Elisée |
author_facet |
Goujard, Laurent Villeneuve, Pierre Baréa, Bruno Lecomte, Jérôme Pina, Michel Claude, Sylvain Le Petit, Jean Ferré, Elisée |
author_sort |
Goujard, Laurent |
title |
A spectrophotometric transesterification-based assay for lipases in organic solvent |
title_short |
A spectrophotometric transesterification-based assay for lipases in organic solvent |
title_full |
A spectrophotometric transesterification-based assay for lipases in organic solvent |
title_fullStr |
A spectrophotometric transesterification-based assay for lipases in organic solvent |
title_full_unstemmed |
A spectrophotometric transesterification-based assay for lipases in organic solvent |
title_sort |
spectrophotometric transesterification-based assay for lipases in organic solvent |
url |
http://agritrop.cirad.fr/547276/ http://agritrop.cirad.fr/547276/1/document_547276.pdf |
work_keys_str_mv |
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