Evaluation of red chicory extract as a natural antioxidant by pure lipid oxidation and yeast oxidative stress response as model systems.

The search for renewable and abundant sources of antioxidants has recently focused on agricultural byproducts, especially promising due to their natural origins and low costs. In particular, plant raw materials are sources of important compounds such as dietary fiber, carotenoids, tocopherols, and polyphenolics, which are mostly discarded during harvesting and processing. Among these vegetal crops, red chicory is attractive because of the large quantity of its byproducts (residues as leaves and stems); moreover, there is no information on its role as a food and feed ingredient. In this study, red chicory leaf residue was evaluated as a natural substitute for synthetic antioxidants for the food and feed industry. After lyophilization, a red chicory extract (RC) was characterized for its phenolic profile and its oxidative stability as compared to BHT. RC was shown to reduce lipid peroxidation of different oils in the Rancimat test. In addition, the antioxidant property of RC was studied in a model system by evaluating the Saccharomyces cerevisiae response to oxidative stress by means of gene expression. In this analysis, the RC extract, added to the yeast culture prior to oxidative stress induction, exhibited a pleiotropic protective effect on stress responsive genes.

Antibrowning potential of Brassicacaea processing water.

The effects of Brassicacaea processing water on polyphenol oxidases from different plant sources were investigated. Results showed that processing water (PW) prepared by cooking young Brassicacaea leaves with water has the capacity to inhibit both commercial tyrosinase and plant polyphenol oxidases. PW was freeze-dried (LPW) in an attempt to increase its utility and improve its efficiency at inhibition of polyphenol oxidases, however this resulted in a loss of activity, probably due to the stresses of the freezing and/or drying processes. The addition of PW to ascorbic acid resulted in complete inhibition of grape polyphenol oxidase, suggesting that the combination of the two antibrowning agents may be a promising tool to control this enzymatic activity.

3,4-Dihydroxyphenylalanine gel diffusion assay for polyphenol oxidase quantification.

We have developed a simple, inexpensive plate assay to detect and quantify polyphenol oxidase (PPO) activity from different origins. The logarithm of enzyme activity is linearly correlated with the diameter of the dark, l-3,4-dihydroxyphenylalanine (l-DOPA) oxidized circles produced in the gel, thereby allowing quantification of PPO. Moreover, precision and high reproducibility of the assay were confirmed by statistical analysis.

Deriphat 2-DE to visualize polyphenol oxidase in Moscato and Prosecco grapes.

The Deriphat 2-DE was used to visualize polyphenol oxidase (PPO) isoforms of Moscato and Prosecco grape extracts, partially purified and characterized. Catecholase has similar values in the two varieties, whereas Moscato cresolase data are almost 54% higher. In the first dimension, the PPO of both varieties may be detected by SDS-PAGE, but native PAGE (N-PAGE) gave negative results. For this reason, the samples were solubilized in the zwitteronic detergent Deriphat, which was also included in the gel and the cathodic buffer. Deriphat migrated together with the cathodic buffer, maintaining protein solubility and revealing the PPO profiles of Moscato and Prosecco extracts in native conditions. The combination of Deriphat-PAGE (D-PAGE) and SDS-PAGE (2-DE) also resulted in improved separation efficiency in resolving PPO and specialized stains in evaluating PPO activities. The control, represented by IEF for the first-dimensional separation, had a lower number of spots, demonstrating the higher capacity of Deriphat 2-DE to isolate PPO isoforms from grape extracts. The Deriphat 2-DE method described here is simple but powerful, and the resulting information will be a useful tool for further proteomic research.