Title : A sn-2 regioselective lipase with cis-fatty acid preference from Cordyceps militaris: Biochemical characterization and insights into its regioselective mechanism - Lee_2024_Int.J.Biol.Macromol__134013 |
Author(s) : Lee J , Choi Y , Kim T , Chang PS |
Ref : Int J Biol Macromol , :134013 , 2024 |
Abstract :
Lipase with unique regioselectivity is an attractive biocatalyst for elaborate lipid modification. However, the excavation of novel sn-2 regioselective lipases is difficult due to their scarcity in nature, with Candida antarctica lipase A (CALA) being the pronouncedly reported one. Here, we identified a novel CALA-like lipase from Cordyceps militaris (CACML7) via in silico mining. Through chiral-phase high-performance liquid chromatography, we determined that CACML7 displays sn-2 regioselectivity (>68 %) as does CALA, but exhibits distinctive chain length selectivity and bias against unsaturated fats. Notably, the curvature of the acyl-binding tunnel was expected to contribute to the 2.2-fold higher preference for cis-fatty acid (C18:1, cis-delta(9)) over trans-fatty acid (C18:1, trans-delta(9)) unlike trans-active CALA. Random pose docking of trioleoylglycerol (TOG) into the active site of a lid-truncated mutant of CACML7 revealed that TOG accepts a tuning fork conformation, of which the precise positioning of the reactive ester group towards the catalytic center was only favorable via sn-2 binding mode. The unique active site morphology, which we refer to as an "acyl-binding tunnel with a narrow entrance," may contribute to the sn-2 regioselectivity of CACML7. Our data provide an attractive model to better understand the mechanism underlying sn-2 regioselectivity. |
PubMedSearch : Lee_2024_Int.J.Biol.Macromol__134013 |
PubMedID: 39032883 |
Gene_locus related to this paper: cormi-a0a2h4s804 , cormi-a0a2h4sba8 , cormi-a0a2h4sdc8 , cormi-a0a2h4sed1 , cormi-a0a2h4smd8 , cormi-a0a2h4snx0 , cormi-a0a2h4sqq8 , cormm-g3j319 , cormm-g3jbi5 , cormm-g3jpg8 |
Lee J, Choi Y, Kim T, Chang PS (2024)
A sn-2 regioselective lipase with cis-fatty acid preference from Cordyceps militaris: Biochemical characterization and insights into its regioselective mechanism
Int J Biol Macromol
:134013
Lee J, Choi Y, Kim T, Chang PS (2024)
Int J Biol Macromol
:134013