Self-assembled bifunctional nanoflower-enabled CRISPR/Cas biosensing platform for dual-readout detection of Salmonella enterica.

Sensitive detection and point-of-care test of bacterial pathogens is of great significance in safeguarding the public health worldwide. Inspired by the characteristics of horseradish peroxidase (HRP), we synthesized a hybrid nanoflower with peroxidase-like activity via a three-component self-assembled strategy. Interestingly, the prepared nanozyme not only could act as an alternative to HRP for colorimetric biosensing, but also function as a unique signal probe that could be recognized by a pregnancy test strip. By combining the bifunctional properties of hybrid nanoflower, isothermal amplification of LAMP, and the specific recognition and non-specific cleavage properties of CRISPR/Cas12a system, the dual-readout CRISPR/Cas12a biosensor was developed for sensitive and rapid detection of Salmonella enterica. Moreover, this platform in the detection of Salmonella enterica had limits of detection of 1 cfu/mL (colorimetric assay) in the linear range of 101-108 cfu/mL and 102 cfu/mL (lateral flow assay) in the linear range of 102-108 cfu/mL, respectively. Furthermore, the developed biosensor exhibited good recoveries in the spiked samples (lake water and milk) with varying concentrations of Salmonella enterica. This work provides new insights for the design of multifunctional nanozyme and the development of innovative dual-readout CRISPR/Cas system-based biosensing platform for the detection of pathogens.

Robust and recyclable magnetic nanobiocatalysts for extraction of anthocyanin from black rice.

Anthocyanins, which are natural pigments and nutraceuticals, can be extracted from plant materials using enzyme-assisted methods. However, the enzymes used are often expensive, fragile, and hard to recover/reuse. In this study, cellulase and α-amylase were immobilized on amino-functionalized magnetic nanoparticles to prepare a magnetic nanobiocatalyst. The enzymes in this nanobiocatalyst exhibited higher stability and greater catalytic activity than free enzymes, including good thermal stability (50 to 70℃) and pH stability (pH 4.5-7.5). Nanobiocatalyst efficacy was demonstrated by extracting anthocyanins from black rice, with a maximum yield of 266 mg anthocyanin/100 g black rice obtained. After six reuse cycles, cellulase and α-amylase retained around 70% and 64% of their activity, respectively. Immobilization also increased their reusability. In summary, a novel magnetic nanobiocatalyst was developed for extracting anthocyanins from black rice, which may also have other applications within the food industry.

Inhibition of Lipid and Protein Oxidation in Whey-Protein-Stabilized Emulsions Using a Natural Antioxidant: Black Rice Anthocyanins.

The food industry is exploring the natural environment to identify botanical extracts that can be used as functional ingredients that can replace synthetic ingredients in foods. In the present study, the ability of black rice anthocyanins as natural antioxidants to inhibit both lipid and protein oxidation in protein-stabilized oil-in-water emulsions was examined. Whey-protein-stabilized emulsions were prepared containing 0, 0.02, 0.04, and 0.06% (w/v) anthocyanins, and then the impact of this plant-based extract on their physical and chemical stabilities was evaluated. The addition of the anthocyanins improved the physical stability of the emulsions in a dose-dependent manner by inhibiting droplet aggregation during storage (35 °C for 5 days). The anthocyanins also exhibited good antioxidant activity in a dose-dependent manner, as seen by their capacity for inhibiting both lipid oxidation (reduced lipid hydroperoxides and malondialdehyde) and protein oxidation (reduced carbonyl and Schiff base formation, intrinsic fluorescence loss, and molecular weight changes). Black rice anthocyanins may therefore be an effective botanical extract for improving the stability of protein-stabilized food emulsions by inhibiting oxidative reactions.