Xing_2024_Enzyme.Microb.Technol_182_110532

GDSL-type esterases are promising biocatalysts for the food and pharmaceutical industries. Here, a GDSL-type esterase from Aspergillus niger CCTCC No. M2012538 (INANE1) was expressed and purified in Pichia pastoris GS115, and its catalytic performances were evaluated, including the synthesis of cinnamyl acetate and deacetyl-7-aminocephalosporanic acid (D-7-ACA). In addition, molecular docking and molecular dynamics simulations analyzed INANE1's substrate specificity. The substrate specificity profile indicated the recombinant esterase (rINANE1) was an acetylesterase with high specificity for p-nitrophenyl acetate (p-NPA). The rINANE1 exhibited maximum activity at pH 8.0 and 35 degreesC, where K(m) and V(max) were calculated as 0.13+/-0.03 mM and 22.56 +/- 0.32 micromoL/min/mg, respectively. The yield of cinnamyl acetate of about 85 % was achieved in 24 h. The conversion rate of 7-aminocephalosporanic acid (7-ACA) could reach 92.71 +/- 1.78 % at 25 degreesC and 2.5 h. Moreover, the INANE1 structure model, molecular docking, and molecular dynamics simulation demonstrated that the pocket of the catalytic triad Ser34, Asn267, and His270 could only accommodate p-NPA. INANE1 may be the first fungi esterase with cinnamyl acetate synthetic activity and 7-ACA hydrolysis activity. Therefore, INANE1 would be a promising enzyme with industrial values.