Homologous yeast lipases/acyltransferases exhibit remarkable cold-active properties

Abstract

Lipases/acyltransferases catalyse acyltransfer to various nucleophiles preferentially to hydrolysis even in aqueous media with high thermodynamic activity of water (a w >0.9). Characterization of hydrolysis and acyltransfer activities in a large range of temperature (5 to 80 °C) of secreted recombinant homologous lipases of the Pseudozyma antarctica lipase A superfamily (CaLA) expressed in Pichia pastoris, enlighten the exceptional cold-activity of two remarkable lipases/acyltransferases: CpLIP2 from Candida parapsilosis and CtroL4 from Candida tropicalis. The activation energy of the reactions catalysed by CpLIP2 and CtroL4 was 18-23 kJ mol(-1) for hydrolysis and less than 15 kJ mol(-1) for transesterification between 5 and 35 °C, while it was respectively 43 and 47 kJ mol(-1) with the thermostable CaLA. A remarkable consequence is the high rate of the reactions catalysed by CpLIP2 and CtroL4 at very low temperatures, with CpLIP2 displaying at 5 °C 65 % of its alcoholysis activity and 45 % of its hydrolysis activity at 30 °C. These results suggest that, within the CaLA superfamily and its homologous subgroups, common structural determinants might allow both acyltransfer and cold-active properties. Such biocatalysts are of great interest for the efficient synthesis or functionalization of temperature-sensitive lipid derivatives, or more generally to lessen the environmental impact of biocatalytic processes.

The 3D model of the lipase/acyltransferase from Candida parapsilosis, a tool for the elucidation of structural determinants in CAL-A lipase superfamily

“…Another interesting feature of CpLIP2 and its homologs CtroL4 and CaLIP4 is their high specific activity at low temperatures, with for example CpLIP2 displaying at 5°C 65% of its alcoholysis activity at 30°C, allowing their classification as cold-active lipases contrary to their more distant homolog CAL-A (31% identity with CpLIP2) [51]. Within the CAL-A family, lipases thus exhibit differences in their catalytic properties such as acyltransfer ability, substrate specificity, thermostability and activity at low temperature [23,51,52].…”

Revealing the Roles of Subdomains in the Catalytic Behavior of Lipases/Acyltransferases Homologous to CpLIP2 through Rational Design of Chimeric Enzymes

“…As observed before for the characterized lipases/acyltransferases CpLIP2 and CtroL4, enzymes of this family do not seem to share the exceptional thermostability of CAL‐A . Despite their high sequence identity, CduLAc has a mesophilic profile, with an activation energy of 34 kJ mol −1 (Table ), whereas CalLAc8 is a cold‐active enzyme, with E A =15 kJ mol −1 (similar to CpLIP2, as described by Neang et al) . The exchanges of cap and C‐terminal flap did not have an impact on the cold‐active property of CalLAc8 (CalLAc8_cap and CalLAc8_ct) or on the thermophilic characteristic of CAL‐A (CAL‐A_ct).…”

“…CalLAc8 and CduLAc exhibited heat lability, with a significant decrease of activity above 20 and 40 °C respectively, and 4 % maximal activity at 80 °C after 15 min (Figure ). As observed before for the characterized lipases/acyltransferases CpLIP2 and CtroL4, enzymes of this family do not seem to share the exceptional thermostability of CAL‐A . Despite their high sequence identity, CduLAc has a mesophilic profile, with an activation energy of 34 kJ mol −1 (Table ), whereas CalLAc8 is a cold‐active enzyme, with E A =15 kJ mol −1 (similar to CpLIP2, as described by Neang et al) .…”

“…More recently, the use unusual lipases that preferentially catalyze transesterification reactions in water and under mild conditions has appeared very promising to reach higher reaction yields and rates under “greener” conditions . With this aim, we reported a group of lipases/acyltransferases homologous to CpLIP2 from Candida parapsilosis . CpLIP2 was the first lipase/acyltransferase to be described .…”

Produktion von Peroxyoktansäure mittels immobilisierter CalB: Realisierbarkeitsstudie im einphasigen Stoffsystem mit graduellem Wasserstoffperoxid‐Dosing

“…set to 47 kJ mol -1 according to literature values, which were in the range of 45-47 kJ mol -1 for the chemo-enzymatic epoxidation of dodecene [38]) and 39-47 kJ mol -1 for transesterifications [195,196]. However, no information was given on the activation energy constraint of a reverse reaction.…”