Comprehensive Analysis of a Yeast Lipase Family in the Yarrowia Clade
| Autor: | Muchalin Meunchan, Hugo Devillers, Jean-Marc Nicaud, Cécile Neuvéglise, Stéphanie Michely, Alain Marty |
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| Přispěvatelé: | Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Department of Biochemistry, Faculty of Science, Masaryk University [Brno] (MUNI), MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, French Government [ANR-001-01], INRA through the AIP-Bioressources YALIP project, Thailand Research Fund (TRF), Khon Kaen University through Royal Golden Jubilee PhD program [PHD/0282/2549], French Minister of Research via the ABIES doctoral school, Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Masaryk University, Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Meunchan, Muchalin, Michely, Stéphanie, Marty, Alain, Neuvéglise, Cécile |
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
[SDV]Life Sciences [q-bio]
Molecular Sequence Data lcsh:Medicine Gene Expression Yarrowia Sequence alignment yeast Synteny Substrate Specificity Evolution Molecular Fungal Proteins Industrial Microbiology Phylogenetics Amino Acid Sequence Lipase lcsh:Science levure Phylogeny Candida Comparative genomics Fungal protein Multidisciplinary Binding Sites biology Sequence Homology Amino Acid Hydrolysis lcsh:R Stereoisomerism biology.organism_classification Yeast Butyrates Biochemistry biology.protein Mutagenesis Site-Directed lcsh:Q Transcriptome Sequence Alignment Research Article Protein Binding |
| Zdroj: | PLoS ONE PLoS ONE, Public Library of Science, 2015, 10 (11), pp.e0143096. ⟨10.1371/journal.pone.0143096⟩ PLoS ONE, Vol 10, Iss 11, p e0143096 (2015) PLoS ONE, 2015, 10 (11), pp.e0143096. ⟨10.1371/journal.pone.0143096⟩ Plos One 11 (10), e0143096. (2015) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0143096⟩ |
| Popis: | Lipases are currently the subject of intensive studies due to their large range of industrial applications. The Lip2p lipase from the yeast Yarrowia lipolytica (YlLIP2) was recently shown to be a good candidate for different biotechnological applications. Using a combination of comparative genomics approaches based on sequence similarity, synteny conservation, and phylogeny, we constructed the evolutionary scenario of the lipase family for six species of the Yarrowia clade. RNA-seq based transcriptome analysis revealed the primary role of LIP2 homologues in the assimilation of different substrates. Once identified, these YlLIP2 homologues were expressed in Y. lipolytica. The lipase Lip2a from Candida phangngensis was shown to naturally present better activity and enantioselectivity than YlLip2. Enantioselectivity was further improved by site-directed mutagenesis targeted to the substrate binding site. The mono-substituted variant V232S displayed enantioselectivity greater than 200 and a 2.5 fold increase in velocity. A double-substituted variant 97A-V232F presented reversed enantioselectivity, with a total preference for the R-enantiomer. |
| Databáze: | OpenAIRE |
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