A high-throughput differential scanning fluorimetry method for rapid detection of thermal stability and iron saturation in lactoferrin.

Thermal stability and iron saturation of lactoferrin (LF) are of great significance not only for the evaluation of the biological activities of LF but also for the optimization of the isolation and drying process parameters. Differential scanning calorimetry (DSC) is a well-established and efficient method for thermal stability and iron saturation detection in LF. However, multiple DSC measurements are typically performed sequentially, thus time-consuming and low throughput. Herein, we introduced the differential scanning fluorimetry (DSF) approach to overcome such limitations. The DSF can monitor LF thermal unfolding with a commonly available real-time PCR instrument and a fluorescent dye (SYPRO orange or Glomelt), and the measured melting temperature of LF is consistent with that determined by DSC. On the basis of that, a new quantification method was established for determination of iron saturation levels using the linear correlation of the degree of ion saturation of LF with DSF measurements. Such DSF method is simple, inexpensive, rapid (<15 min), and high throughput (>96 samples per experiment), and provides a valuable alternative tool for thermal stability detection of LF and other whey proteins.

Characterization of fermented pomegranate juice: ACE inhibitory activity under in vitro digestion, antioxidant capacity, phenolics composition, chemical properties and sensory evaluation.

Consuming pomegranate juice (PJ) is beneficial for hypertensive regulation because of the phenolic compounds in PJ and their inhibitory activity on angiotensin-I-converting enzyme (ACE). To better utilize bioactive function of food, microorganism fermentation has been adopted to alter phenolic metabolism. This study confirms that even under in vitro digestion, fermented PJ (FPJ) maintains higher ACE inhibitory activity than that of PJ. The main phenolic compounds in PJ were compared either under fermentation or in vitro digestion. This study finds that fermentation promotes antioxidant capacity of PJ. The chemical properties of FPJ are evaluated and the corresponding relationship with bioactivities is analyzed. A sensory evaluation comparison is conducted between FPJ and PJ, furnishing interesting information for consumers. This study highlights the relationship between ACE inhibitory activity of PJ and phenolic composition under fermentation and in vitro digestion, providing novel insights for diet regulation of phenolic-rich FPJ in ACE inhibition therapy. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01388-w.