Carrier-Bound And Carrier-Free Immobilization Of Type A Feruloyl Esterase from Aspergillus niger: Searching For An Operationally Stable Heterogeneous Biocatalyst For The Synthesis Of Butyl Hydroxycinnamates.

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Carrier-Bound And Carrier-Free Immobilization Of Type A Feruloyl Esterase from Aspergillus niger: Searching For An Operationally Stable Heterogeneous Biocatalyst For The Synthesis Of Butyl Hydroxycinnamates.

J Biotechnol. 2020 Apr 16;:

Authors: Grajales-Hernández DA, Velasco-Lozano S, Armendáriz-Ruiz MA, Rodríguez-González JA, Camacho-Ruíz RM, Asaff-Torres A, López-Gallego F, Mateos-Díaz JC

Abstract
Feruloyl esterases synthesize butyl hydroxycinnamates, molecules possessing interesting biological properties, nonetheless, they exhibit a low stability under synthesis conditions in organic solvents, restricting its use. To enhance its operational stability in synthesis, we immobilized type A feruloyl esterase from Aspergillus niger (AnFAEA) using several carrier-bound and carrier-free strategies. The most active biocatalysts were: 1) AnFAEA immobilized on epoxy-activated carriers (protein load of 0.6 mgenzyme x mg-1carrier) that recovered 91% of the initial hydrolytic activity, and 2) AnFAEA aggregated and cross-linked in the presence of 5 mg of BSA and 15 mM of glutaraldehyde (AnFAEA-amino-CLEAs), which exhibited 385% of its initial hydrolytic activity; both using 4-nitrophenyl butyrate as substrate. The AnFAEA-amino-CLEAs were 12.7 times more thermostable at 60 °C than the AnFAEA immobilized on epoxy-activated carrier, thus AnFAEA-amino-CLEAs were selected for further characterization. Interestingly, during methyl sinapate hydrolysis (pH 7.2 and 30 °C), AnFAEA-amino-CLEAs KM was 15% higher, while during butyl sinapate synthesis the KM was reduced in 63%, both compared with the soluble enzyme. The direct esterification of butyl sinapate at solvent free conditions using sinapic acid 50 mM, reached 95% conversion after 24 h employing AnFAEA-amino-CLEAs, which could be used for 10 cycles without significant activity losses, demonstrating their outstanding operational stability.

PMID: 32305629 [PubMed – as supplied by publisher]

Source: Industry