Abstract
The enzymatic synthesis of alkyl ferulates is an important reaction in cosmetic and pharmaceutical chemistries, since it may allow to expand the biorefinery concept valorizing biomass wastes enriched in ferulic acid. However, robust biocatalysts for that purpose are scarce. Herein, we have immobilized the type A feruloyl esterase from Aspergillus niger (AnFaeA) as cross-linked enzyme aggregates, employing chitosan as co-feeder (ChCLEAs). High immobilization yields and relative activity recovery were attained in all assessed conditions (> 93%). Furthermore, we enhanced the thermal stability of the soluble enzyme 32-fold. AnFaeA-ChCLEAs were capable to quantitatively perform the solvent-free direct esterification of short- to medium-chain alkyl ferulates (C4-C12) in less than 24 h. By raising the operational temperature to 50 °C, AnFaeA-ChCLEAs transformed 350 mM ferulic acid into isopentyl ferulate with a space-time yield of 46.1 g of product × L-1 × day-1, 73-fold higher than previously reported. The overall sustainability of this alkyl ferulate production bioprocess is supported by the high total turnover number (TTN 7 × 105) and the calculated green metrics (E factor = 30). Therefore, we herein present a robust, efficient, and versatile heterogeneous biocatalyst useful for the synthesis of a wide diversity of alkyl ferulates.
Key points
• CLEAs of feruloyl esterase A from A. niger using chitosan as co-feeder were obtained.
• Microenvironment of the biocatalysts allowed to obtain C1 to C18 alkyl ferulates.
• Biocatalyst at boundary conditions showed a high productivity of 46 g/L day.
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Acknowledgments
Daniel Grajales and Mariana Armendáriz acknowledge scholarships received from CONACyT. Thanks to Dr. Eric Record and Dr. Anthony Levasseur for their kind donation of the A. niger transformant (INRA, Marseille, France). The authors acknowledge the funding of Mexican and Spanish governments.
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The work here presented was funded by the program Atención a Problemas Nacionales (project number 2017-01-6267) granted by CONACyT, Mexico, as well as by the IKERBASQUE and Spanish government (projects BIO2015-69887-R and RTI2018-094398-B-I00)
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JCMD, FLG, and DAG conceived and designed the research; DAG contributed with conceptualization, methodology design, investigation, and writing the original draft; MAR performed investigation, methodology design, and reviewing. SVL contributed by conceptualizing, methodology design, investigation, and review and editing. Finally FLG contributed by obtaining resources, supervision, visualization, review and editing; and JCMD contributed in the conceptualization, methodology design , supervision, project administration, resources, funding acquisition, and review and editing. All authors read and approved the manuscript.
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Grajales-Hernández, D., Armendáriz-Ruiz, M., Velasco-Lozano, S. et al. Chitosan-based CLEAs from Aspergillus niger type A feruloyl esterase: high-productivity biocatalyst for alkyl ferulate synthesis. Appl Microbiol Biotechnol 104, 10033–10045 (2020). https://doi.org/10.1007/s00253-020-10907-2
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DOI: https://doi.org/10.1007/s00253-020-10907-2