Integrating recognition elements with nanomaterials for bacteria sensing.

Pathogenic bacterial contamination is a major threat to human health and safety. In this review, we summarize recent strategies for the integration of recognition elements with nanomaterials for the detection and sensing of pathogenic bacteria. Nanoprobes can provide sensitive and specific detection of bacterial cells, which can be applied across multiple applications and industries.

Immobilization and stabilization of lipase (CaLB) through hierarchical interfacial assembly.

Nanostructure-enabled hierarchical assembly holds promise for efficient biocatalyst immobilization for improved stability in bioprocessing. In this work we demonstrate the use of a hierarchical assembly immobilization strategy to enhance the physicochemical properties and stability of lipase B from Candida antarctica (CaLB). CaLB was complexed with iron oxide nanoparticles followed by interfacial assembly at the surface of an oil-in-water emulsion. Subsequent ring opening polymerization of the oil provided cross-linked microparticles that displayed an increase in catalytic efficiency when compared to the native enzyme and Novozym 435. The hierarchical immobilized enzyme assembly showed no leakage from the support in 50% acetonitrile and could be magnetically recovered across five cycles. Immobilized lipase exhibited enhanced thermal and pH stability, providing 72% activity retention after 24 h at 50 °C (pH 7.0) and 62% activity retention after 24 h at pH 3.0 (30 °C); conditions resulting in complete deactivation of the native lipase.

Transforming food waste: how immobilized enzymes can valorize waste streams into revenue streams.

Food processing generates byproduct and waste streams rich in lipids, carbohydrates, and proteins, which contribute to its negative environmental impact. However, these compounds hold significant economic potential if transformed into revenue streams such as biofuels and ingredients. Indeed, the high protein, sugar, and fat content of many food waste streams makes them ideal feedstocks for enzymatic valorization. Compared to synthetic catalysts, enzymes have higher specificity, lower energy requirement, and improved environmental sustainability in performing chemical transformations, yet their poor stability and recovery limits their performance in their native state. This review article surveys the current state-of-the-art in enzyme stabilization & immobilization technologies, summarizes opportunities in enzyme-catalyzed valorization of waste streams with emphasis on streams rich in mono- and disaccharides, polysaccharides, lipids, and proteins, and highlights challenges and opportunities in designing commercially translatable immobilized enzyme systems towards the ultimate goals of sustainable food production and reduced food waste.

Stabilization of Lipase in Polymerized High Internal Phase Emulsions.

Candida antarctica lipase B is stabilized in a porous, high internal phase emulsion (HIPE) of polydicyclopentadiene to enable biocatalytic waste stream upcycling. The immobilized lipase is subjected to thorough washing conditions and tested for stability in extreme environments and reusability. A porous internal microstructure is revealed through scanning electron microscopy. After preparation, lipase activity increased to 139 ± 9.7% of its original activity. After 10 cycles of reuse, immobilized lipase retains over 50% activity. Immobilized lipase retains activity after 24 h of exposure to temperatures ranging from 20 to 60 °C and pH values of 3, 7, and 10. In the most extreme environments tested, lipase retained 42.8 ± 21% relative activity after exposure to 60 °C and 49.4 ± 16% relative activity after exposure to pH 3. Polymerized HIPEs stabilize lipase and, thus, extend its working range. Further synthesis optimization has the potential to increase enzyme stability, immobilization efficiency, and uniformity. The reported hierarchical stabilization technique shows promise for use of immobilized lipase in non-ideal, industrially relevant conditions.

Influence of Hierarchical Interfacial Assembly on Lipase Stability and Performance in Deep Eutectic Solvent.

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.