Multicompartment calcium alginate microreactors to reduce substrate inhibition in enzyme cascade reactions.

The formation of macromolecularly enriched condensates through associative or segregative liquid-liquid phase separation phenomena is known to play a central role in controlling various cellular functions in nature. The potential to spatially and temporally modulate multistep chemical reactions and pathways has inspired the use of phase-separated systems for the development of various synthetic colloidal micro- and nanoreactor systems. Here, we report a rational and synthetically minimal design strategy to emulate intended spatiotemporal functions in morphologically intricate and structurally defined calcium alginate hydrogel microreactors possessing multicompartmentalized internal architectures. Specifically, we implement a thermal phase separation protocol to achieve fine-control over liquid-liquid phase separation inside complex aqueous emulsion droplet templates that are loaded with hydrophilic polymer mixtures. Subsequent gelation of alginate-containing droplet templates using a novel freeze-thaw approach that can be applied to both scalable batch production or more precise microfluidic methods yields particle replicas, in which subcompartmentalized architectures can be retained. Larger active components can be enriched in the internal compartments due to their preferential solubility, and we show that selective sequestration of enzymes serves to create desired microenvironments to control and tune the reaction kinetics of a multistep enzyme cascade by reducing their mutual interference. This demonstration of mitigating substrate inhibition that is based primarily on optimizing the multicompartmentalized hydrogel particle morphology offers new opportunities for the simple and synthetically-minimal batch generation of hydrogel-based synthesis microreactors.

CO2-responsive Pickering emulsions stabilized by soft protein particles for interfacial biocatalysis.

Pickering emulsions are emulsions stabilized by colloidal particles and serve as an excellent platform for biphasic enzymatic catalysis. However, developing simple and green strategies to avoid enzyme denaturation, facilitate product separation, and achieve the recovery of enzyme and colloidal particle stabilizers is still a challenge. This study aimed to report an efficient and sustainable biocatalysis system via a robust CO2/N2-responsive Pickering oil-in-water (o/w) emulsion stabilized solely by pure sodium caseinate (NaCas), which was made naturally in a scalable manner. The NaCas-stabilized emulsion displayed a much higher reaction efficiency compared with conventional CO2/N2-responsive Pickering emulsions stabilized by solid particles with functional groups from polymers or surfactants introduced to tailor responsiveness, reflected by the fact that most enzymes were transferred and enriched at the oil-water interface. More importantly, the demulsification, product separation, and recycling of the NaCas emulsifier as well as the enzyme could be facilely achieved by alternatively bubbling CO2/N2 more than 30 times. Moreover, the recycled enzyme still maintained its catalytic activity, with a conversion yield of more than 90% after each cycle, which was not found in any of the previously reported CO2-responsive systems. This responsive system worked well for many different types of oils and was the first to report on a protein-based CO2/N2-responsive emulsion, holding great promise for the development of more sustainable, green chemical conversion processes for the food, pharmaceutical, and biomedical industries.

Growth of Au nanoparticles on phosphorylated zein protein particles for use as biomimetic catalysts for cascade reactions at the oil-water interface.

Chemo-enzymatic cascade processes are invaluable due to their ability to rapidly construct high-value products from available feedstock chemicals in a one-pot relay manner. However, they have proven to be challenging because of the mutual inactivation of both catalysts. A conceptually novel strategy based on Pickering interfacial catalysis (PIC) is proposed here to address this challenge. This study aimed to construct a protein-stabilized Pickering system for biphasic cascade catalysis, enabled by phosphorylated zein nanoparticles (ZCPOPs) immobilized in gold nanoparticles (Au NCs). Ultra-small Au NCs, 1-2 nm in diameter, were integrated into ZCPOPs at room temperature. Then, the as-synthesized ZCPOPs-Au NCs were used to stabilize the oil-in-water (o/w) Pickering emulsion. Besides their excellent catalytic activity and recycling ability in a variety of oil phases, ZCPOPs-Au NCs possess unpredictable catalytic activity and exhibit mimicking properties of horseradish peroxidase. Particularly, the cascade reaction is well achieved using a metal catalyst and a biocatalyst at the oil-water interface. The result showed that such a combination of chemo- and biocatalysis improved the catalytic yield more than two times compared with that of sole metal catalysis. This study opened a new avenue to design nanomaterials using the combination of chemo- and biocatalysis in a Pickering emulsion system for multistep syntheses.

Sodium caseinate as a particulate emulsifier for making indefinitely recycled pH-responsive emulsions.

pH-responsive emulsions are one of the simplest and most readily implementable stimuli-responsive systems. However, their practical uses have been greatly hindered by cyclability. Here, we report a robust pH-responsive emulsion prepared by utilizing pure sodium caseinate (NaCas) as the sole emulsifier. We demonstrate that the emulsification/demulsification of the obtained NaCas-stabilized emulsion can be triggered by simply changing the pH value over 100 cycles, which has never been observed in any protein-stabilized emulsion system. The NaCas-stabilized emulsion maintains its pH-responsive properties even in a saturated salt solution (NaCl ∼ 6.1 M) or seawater. We illustrate how NaCas functions in pH-responsive emulsions and show that when conventional nanoparticles such as zein protein or bare SiO2 particles were coated with a layer of NaCas, the resulting formulated emulsions could be switched on and off over 10 cycles. The unique properties of NaCas thus enable the engineering of conventional Pickering emulsions to pH-responsive Pickering emulsions. Finally, we have integrated catalytically active gold (Au) nanoclusters (NCs) into the NaCas protein and then utilized them to produce emulsions. Remarkably, these NaCas-Au NCs assembled at the oil-water interface exhibited excellent catalytic activity and cyclability, not only in aqueous solution, but also in complicated seawater environments.

Biocycle Fermentation Based on Lactic Acid Bacteria and Yeast for the Production of Natural Ethyl Lactate.

Ethyl lactate is widely used in food and pharmaceutical industries, but the complexity of the synthesis process, in particular, involving the addition of organic solvents, hinders its application. Here, we report a natural green strategy to produce ethyl lactate by exploiting the synergistic fermentation of lactic acid bacteria and ester-producing microbes using biomass as a substrate. Interestingly, it is worth noting that the conjugate fermentation has a higher ethyl lactate yield (3.05 g/L) compared to the mixed fermentation (1.32 g/L). The ester production capacity was increased by 2.3 times. These entire processes require only the addition of biomass without introducing any organic solvent. In addition, the obtained catalytic esterification system can reuse the ester-producing microbes by simple centrifugation and maintain over seven cycles of catalysis while it retained a high activity. We firmly believe that the results of this study will provide new ideas for achieving sustainable green production of natural ethyl lactate.

pH-Responsive Emulsions with ß-Cyclodextrin/Vitamin E Assembled Shells for Controlled Delivery of Polyunsaturated Fatty Acids.

Lipid-based delivery systems (LBDSs) are widely applied in pharmaceuticals and health care because of the increased bioavailability of lipophilic components when they are coadministered with high-fat meals. However, how to accurately control their in vivo release and stability is still challenging. Here, after introducing the simple esterification and coprecipitation, we created the dual-functional composite ODS-ß-CD-VE by the coassembly of ß-cyclodextrin (ß-CD), octadecenyl succinic anhydride (ODSA), and vitamin E (VE). The resulting dual-functional particle presented a uniform sheetlike shape and nanometer size. In addition, its chemical structure was clarified in detail via nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). Benefiting from the antioxygenation of VE, lipid oxidation in the ODS-ß-CD-VE-stabilized Pickering emulsion was effectively inhibited. Meanwhile, pH-induced protonation/deprotonation of carboxyl groups guaranteed that the emulsions kept steady at pH ≤4 but were unsteady under neutral conditions. In this way, the lipids contained in the emulsion were protected from gastric juice and then digested and accurately released as n-3 polyunsaturated fatty acids (PUFA) in the simulated intestine environment. This strategy sheds some light on the rational and efficient construction of LBDSs for nutrient supplements and even pharmaceuticals in a living digestive tract.

Protein-Based Pickering High Internal Phase Emulsions as Nutraceutical Vehicles of and the Template for Advanced Materials: A Perspective Paper.

Pickering high internal phase emulsions (HIPEs) are normally highly concentrated emulsions stabilized by colloidal particles with a minimum internal phase volume fraction of 0.74. They have received considerable attention in many fields, including pharmaceuticals, tissue engineering, foods, and personal care products. The aim of this perspective is to update the current knowledge on the field of protein-based Pickering HIPEs, emphasizing those aspects that need to be explored and clarified. Research progress in constructing HIPEs by protein-type colloid particles and promising research trends in basic research and potential applications were highlighted. Promising studies in this field include (1) clarifying bioavailability and evolution of activity of active ingredients in Pickering HIPEs by oral administration, (2) constructing a Pickering interfacial catalysis platform using protein colloidal particles, and (3) expanding the emerging applications of Pickering HIPEs in fields, such as partially hydrogenated oil replacers, probiotic encapsulation, and the template for porous materials.

Modulation of Cyclodextrin Particle Amphiphilic Properties to Stabilize Pickering Emulsion.

Cyclodextrins have been proven to form complexes with linear oil molecules and stabilize emulsions. Amphiphilic properties of cyclodextrin particles were modulated through esterification reaction between ß-cyclodextrin (ß-CD) and octadecenyl succinic anhydride (ODSA) under alkaline conditions. ODS-ß-CD particles with degree of substitution (DS) of 0.003, 0.011, and 0.019 were obtained. The introduced hydrophobic long chain that was linked within ß-CD cavity led to the change of ODS-ß-CD in terms of morphological structure, surface charge density, size, and contact angle, upon which the properties and stability of the emulsions stabilized by ODS-ß-CD were highly dependent. The average diameter of ODS-ß-CD particles ranged from 449 to 1484 nm. With the DS increased from 0.003 to 0.019, the contact angle and absolute zeta potential value of these ODS-ß-CD particles improved from 25.7° to 47.3° and 48.1 to 62.8 mV, respectively. The cage structure of ß-CD crystals was transformed to channel structure, then further to amorphous structure after introduction of the octadecenyl succinylation chain. ODS-ß-CD particles exhibited higher emulsifying ability compared to ß-CD. The resulting Pickering emulsions formed by ODS-ß-CD particles were more stable during storage. This study investigates the ability of these ODS-ß-CD particles to stabilize oil-in-water emulsions with respect to their amphiphilic character and structural properties.