Enzymatic interfacial conversion of acylglycerols in Pickering emulsions stabilized by hydrogel microparticles.

HYPOTHESIS: A critical challenge in the enzymatic conversion of acylglycerols is the limited exposure of the enzyme dissolved in the aqueous solution to the hydrophobic substrate in the oil phase. Positioning the enzyme in a microenvironment with balanced hydrophobicity and hydrophilicity in Pickering emulsion will facilitate the acylglycerol-catalyzing reactions at the interface between the oil and liquid phases. EXPERIMENTS: In this work, to overcome the challenge of biphasic catalysis, we report a method to immobilize enzymes in polyethylene glycol (PEG)-based hydrogel microparticles (HMPs) at the interface between the oil and water phases in Pickering emulsion to promote the enzymatic conversion of acylglycerols. FINDINGS: 3 wt% of HMPs can stabilize the oil-in-water Pickering emulsion for at least 14 days and increase the viscosity of emulsions. Lipase-HMP conjugates showed significantly higher hydrolytic activity in Pickering emulsion; HMP-immobilized lipase SMG1 showed an activity about three times that of free lipase SMG1. Co-immobilization of a lipase and a fatty acid photodecarboxylase from Chlorella variabilis (CvFAP) in Pickering emulsion enables light-driven cascade conversion of triacylglycerols to hydrocarbons, transforming waste oil to renewable biofuels in a green and sustainable approach. HMPs stabilize the Pickering emulsion and promote interfacial biocatalysis in converting acylglycerols to renewable biofuels.

Synthetic Multienzyme Assemblies for Natural Product Biosynthesis.

In nature, enzymes that catalyze sequential reactions are often assembled as clusters or complexes. The formation of multienzyme complexes, or metabolons, brings the enzyme active sites into proximity to promote intermediate transfer, decrease intermediate leakage, and streamline the metabolic flux towards the desired products. We and others have developed synthetic versions of metabolons through various strategies to enhance the catalytic rates for synthesizing valuable chemicals inside microbes. Synthetic multienzyme complexes range from static enzyme nanostructures to dynamic enzyme coacervates. Enzyme complexation optimizes the metabolic fluxes inside microbes, increases the product titer, and supplies the field with high-yield microbe strains that are amenable to large-scale fermentation. Enzyme complexes constructed inside microbial cells can be separated as independent entities and catalyze biosynthetic reactions ex vivo; such a feature gains these complexes another name, "synthetic organelles" - new subcellular entities with independent structures and functions. Still, the field is seeking new strategies to better balance dynamicity and confinement and to achieve finer control of local compartmentalization in the cells, as the natural multienzyme complexes do. Industrial applications of synthetic multienzyme complexes for the large-scale production of valuable chemicals are yet to be realized. This review focuses on synthetic multienzyme complexes that are constructed and function inside microbial cells.

Changes in activity, structure and morphology of horseradish peroxidase induced by cold plasma.

Cold plasma is an emerging technology increasingly applied in the agri-food industry. For fruit and vegetables, enzymatic browning is a common phenomenon, causing quality deterioration. The objective of this study was to illustrate the effect of microscale atmospheric-pressure plasma jet (µAPPJ) plasma on the horseradish peroxidase (HRP). Results showed that after plasma treatment for 10 min, the residual activity of HRP was decreased to around 17%, and modification of secondary and tertiary structures were confirmed. The atomic force microscope (AFM) analysis revealed that the aggregation of enzyme protein was enhanced with prolonging treatment time. It was concluded that the activity of HRP could be reduced with destruction of structures and deformation of microstructure induced by µAPPJ plasma. The current study attempted to provide new idea for inhibiting browning enzymes of fruit and vegetables with plasma technology through deeper understanding of the interaction mechanism of plasma active species with enzymes.

Activities and conformation changes of food enzymes induced by cold plasma: A review.

Food endogenous enzymes have impacts on color, texture and flavor of foods during food processing or preservation. Cold plasma is a novel non-thermal food processing technology, which has been extensively studied for contamination elimination and shelf life extension of foods. Particularly, much work has been reported about the effects of cold plasma on enzyme activities and alterations about enzymes conformational structures. It is thus necessary to understand the mechanisms of actions and applications of cold plasma technology in the conformation of food endogenous enzymes. This review focuses on the applications of cold plasma for the inactivation of various endogenous enzymes, including peroxidase, polyphenol oxidase, lysozyme, α-chymotrypsin, alkaline phosphatase, and pectin methylesterase. The activations of several enzymes, such as superoxide dismutase, catalase, and lipase, by cold plasma are also discussed. In addition, this review highlights the transformation of conformational structures including primary and spatial structures induced by chemical reactive species during cold plasma treatments, such as reactive oxygen species and reactive nitrogen species, especially, active sites consisting of prosthetic group and specific amino acids are demonstrated. Both extrinsic and intrinsic factors affecting cold plasma treatments are also described. In general, cold plasma exhibits the ability to activate or inactivate enzymes activities with affecting the conformational structures of enzyme. Further studies should be focused on exploration at molecular level for providing more insight on the interaction mechanism. In addition, equipment and process parameters of cold plasma operation for different fresh food products should be optimized for achieving appropriate control on enzyme variation and obtaining maximum efficiency.