| An ultraviolet spectrophotometric assay for the screening of sn-2-specific lipases using 1,3-O-dioleoyl-2-O-α-eleostearoyl-sn-glycerol as substrate. | |
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MedLine Citation:
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PMID: 22114038 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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In the present study, we propose a continuous assay for the screening of sn-2 lipases by using triacylglycerols (TAGs) from Aleurites fordii seed (tung oil) and a synthetic TAG containing the α-eleostearic acid at sn-2 position and the oleic acid at sn-1 and sn-3 positions (sn-OEO). Each TAG was coated into microplate well and the lipase activity was measured by optical density increase at 272 nm due to transition of α-eleostearic acid from the adsorbed to the soluble state. sn-1,3-regioselective lipases (human pancreatic lipase (HPL) and LIP2 from Yarrowia lipolytica (YLLIP2) and a known sn-2 lipase (Candida antarctica lipase A, CALA) were used to validate this method. Thin layer chromatography analysis of lipolysis products showed that the lipases tested were able to hydrolyze the sn-OEO and the tung oil TAGs, but only CALA hydrolyzed the sn-2 position. The ratio of initial velocities on sn-OEO and tung oil TAGs was used to estimate the sn-2 preference of lipases. CALA was the enzyme with the highest ratio (0.220±015), while HPL and YLLIP2 showed a much lower ratio (0.072±0.026 and 0.038±0.016, respectively). This continuous sn-2 lipase assay is compatible with a high sample throughput and thus can be applied to the screening of sn-2 lipases. |
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Authors:
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Lilia D Mendoza; Jorge A Rodriguez; Julien Leclaire; Gerard Buono; Frederic Fotiadu; Frederic Carriere; Abdelkarim Abousalham |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-11-23 |
Journal Detail:
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Title: Journal of lipid research Volume: - ISSN: 0022-2275 ISO Abbreviation: - Publication Date: 2011 Nov |
Date Detail:
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Created Date: 2011-11-24 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0376606 Medline TA: J Lipid Res Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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UMR 6263 CNRS: Institut des Sciences Moleculaires de Marseille, France; |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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