title: Revealing the Roles of Subdomains in the Catalytic Behavior of Lipases/Acyltransferases Homologous to CpLIP2 through Rational Design of Chimeric Enzymes.
abstract: The lipases/acyltransferases homologous to CpLIP2 of Candida parapsilosis efficiently catalyze acyltransfer reactions in lipid/water media with high water activity (aW>0.9). Two new enzymes of this family, CduLAc from Candida dubliniensis and CalLAc8 from Candida albicans, were characterized. Despite 82 % sequence identity, the two enzymes have significant differences in their catalytic behaviors. In order to understand the roles played by the different subdomains of these proteins (main core, cap and C-terminal flap), chimeric enzymes were designed by rational exchange of cap and C-terminal flap, between CduLAc and CalLAc8. The results show that the cap region plays a significant role in substrate specificity; the main core was found to be the most important part of the protein for acyltransfer ability. Similar exchanges were made with CAL-A from Candida antarctica, but only the C-terminal exchange was successful. Yet, the role of this domain was not clearly elucidated, other than that it is essential for activity.
authors.affiliation: Montpellier Supagro, UMR 1208 IATE, 2 place Viala, 34060, Montpellier cedex 2, France.
authors.forename: Anne-Hélène
authors.initials: AH
authors.lastname: Jan
authors.affiliation: Montpellier Supagro, UMR 1208 IATE, 2 place Viala, 34060, Montpellier cedex 2, France.
authors.forename: Éric
authors.initials: É
authors.lastname: Dubreucq
authors.affiliation: Montpellier Supagro, UMR 1208 IATE, 2 place Viala, 34060, Montpellier cedex 2, France.
authors.forename: Maeva
authors.identifier: 0000-0003-4100-7869
authors.initials: M
authors.lastname: Subileau
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000378:metabolism*; D002175:Candida / Q000201:enzymology* / Q000235:genetics; D002384:Catalysis; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D014867:Water / Q000737:chemistry
chemical_list: D011993:Recombinant Fusion Proteins; D014867:Water; D000217:Acyltransferases; D008049:Lipase
keywords: cap; enzyme catalysis; lipases/acyltransferases; protein engineering; structure-activity relationships