title: Influence of Hierarchical Interfacial Assembly on Lipase Stability and Performance in Deep Eutectic Solvent.
abstract: Hierarchical systems that integrate nano- and macroscale structural elements can offer enhanced enzyme stability over traditional immobilization methods. Microparticles were synthesized using interfacial assembly of lipase B from Candida antarctica with (CLMP-N) and without (CLMP) nanoparticles around a cross-linked polymeric core, to characterize the influence of the hierarchical assembly on lipase stability in extreme environments. Kinetic analysis revealed that the turnover rate (kcat) significantly increased after immobilization. The macrostructure stabilized lipase at neutral and basic pH values, while the nanoparticles influenced stability under acidic pH conditions. Performance of CLMPs was demonstrated by production of sugar ester surfactants in a greener, deep eutectic solvent system (choline chloride and urea). Turnover rate (kcat) and catalytic efficiency (kcat/Km) of the CLMPs decreased following solvent exposure but retained over 60% and 20% activity after 48 h storage at 50 and 60 °C, respectively. CLMP and CLMP-N outperformed the commercially available lipase per unit protein in the production of sugar esters. Improving enzyme performance in greener solvent systems via hierarchical assembly can improve processing efficiency and sustainability for the production of value-added agricultural products.
authors.affiliation: Department of Food Science, University of Massachusetts-Amherst , 102 Holdsworth Way, Amherst, Massachusetts 01003, United States.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Andler
authors.affiliation: Department of Chemistry, University of Massachusetts-Amherst , 379A LGRT, 710 Nt. Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Li-Sheng
authors.initials: LS
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of Massachusetts-Amherst , 379A LGRT, 710 Nt. Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Vincent M
authors.identifier: http://orcid.org/0000-0002-5184-5439
authors.initials: VM
authors.lastname: Rotello
authors.affiliation: Department of Food Science, University of Massachusetts-Amherst , 102 Holdsworth Way, Amherst, Massachusetts 01003, United States.
authors.forename: Julie M
authors.identifier: http://orcid.org/0000-0002-3644-0732
authors.initials: JM
authors.lastname: Goddard
mesh_terms: D002175:Candida / Q000737:chemistry / Q000201:enzymology*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D008049:Lipase
keywords: bioprocessing; deep eutectic solvent; enzyme immobilization; hierarchical assembly; lipase; nanoparticle