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Revealing the Roles of Subdomains in the Catalytic Behavior of Lipases/Acyltransferases Homologous to CpLIP2 through Rational Design of Chimeric Enzymes
Author(s) -
Jan AnneHélène,
Dubreucq Éric,
Subileau Maeva
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600672
Subject(s) - acyltransferases , enzyme , biochemistry , candida parapsilosis , candida albicans , rational design , biology , chemistry , protein engineering , lipase , candida antarctica , stereochemistry , biosynthesis , genetics
Abstract The lipases/acyltransferases homologous to CpLIP2 of Candida parapsilosis efficiently catalyze acyltransfer reactions in lipid/water media with high water activity ( a W >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.