Chlorophylls and carotenoids are natural pigments that are present in our daily diet, especially with the increasing tendency towards more natural and healthy behaviors among consumers. As disturbed antioxidant homeostasis capacities seem to be implicated in the progress of different pathologies, the antioxidant properties of both groups of lipophilic compounds have been studied. The objective of this review was to analyze the state-of-the-art advances in this field. We conducted a systematic bibliographic search (Web of Science™ and Scopus®), followed by a comprehensive and critical description of the results, with special emphasis on highly cited and more recently published research. In addition to an evaluative description of the methodologies, this review discussed different approaches used to obtain a physiological perspective, from in vitro studies to in vivo assays using oxidative biomarkers. From a chemical viewpoint, many studies have demonstrated how a pigment’s structure influences its antioxidant response and the underlying mechanisms. The major outcome is that this knowledge is essential for interpreting new data in a metabolic networks context in the search for more direct applications to health. A promising era is coming where the term “antioxidant” is understood in terms of its broadest significance.

Chlorophyll (chl) breakdown during senescence is an integral part of plant development and leads to the accumulation of colorless catabolites. The loss of green pigment is due to an oxygenolytic opening of the porphyrin macrocycle of pheophorbide (pheide) a followed by a reduction to yield a fluorescent chl catabolite. This step is comprised of the interaction of two enzymes, pheide a oxygenase (PaO) and red chl catabolite reductase. PaO activity is found only during senescence, hence PaO seems to be a key regulator of chl catabolism. Whereas red chl catabolite reductase has been cloned, the nature of PaO has remained elusive. Here we report on the identification of the PaO gene of Arabidopsis thaliana ( AtPaO ). AtPaO is a Rieske-type iron–sulfur cluster-containing enzyme that is identical to Arabidopsis accelerated cell death 1 and homologous to lethal leaf spot 1 (LLS1) of maize. Biochemical properties of recombinant AtPaO were identical to PaO isolated from a natural source. Production of fluorescent chl catabolite-1 required ferredoxin as an electron source and both substrates, pheide a and molecular oxygen. By using a maize lls1 mutant, the in vivo function of PaO, i.e., degradation of pheide a during senescence, could be confirmed. Thus, lls1 leaves stayed green during dark incubation and accumulated pheide a that caused a light-dependent lesion mimic phenotype. Whereas proteins were degraded similarly in wild type and lls1 , a chl-binding protein was selectively retained in the mutant. PaO expression correlated positively with senescence, but the enzyme appeared to be post-translationally regulated as well.

Although there is no legal and clear definition of the term “natural food colorant”, the market trends, and consequently industrial and commercial interest, have turned to foods with added natural pigments. This progressive substitution of artificial colorants has faced chemical complications with some colors, with a lack of stable green hues being one of them. Several strategies have been applied for green color stabilization in processed foods, from the formation of metallochlorophylls to the microencapsulation of green pigments. However, at present, the utilization of green coloring foodstuffs, which are considered an ingredient in the EU, seems to be the more successful solution for the market. Besides those topics, the present review aims to clarify the current confusion between the different chlorophyll compounds that form part of the authorized green food colorants. In this sense, legislations from different countries are compared. Finally, and in line with current concerns, the knowledge gathered so far in relation to the absorption, distribution, metabolism and excretion of all green natural food colorants is reviewed.

A Citrus sinensis spontaneous mutant, navel negra (nan), produces fruit with an abnormal brown-colored flavedo during ripening. Analysis of pigment composition in the wild-type and nan flavedo suggested that typical ripening-related chlorophyll (Chl) degradation, but not carotenoid biosynthesis, was impaired in the mutant, identifying nan as a type C stay-green mutant. nan exhibited normal expression of Chl biosynthetic and catabolic genes and chlorophyllase activity but no accumulation of dephytylated Chl compounds during ripening, suggesting that the mutation is not related to a lesion in any of the principal enzymatic steps in Chl catabolism. Transcript profiling using a citrus microarray indicated that a citrus ortholog of a number of SGR (for STAY-GREEN) genes was expressed at substantially lower levels in nan, both prior to and during ripening. However, the pattern of catabolite accumulation and SGR sequence analysis suggested that the nan mutation is distinct from those in previously described stay-green mutants and is associated with an upstream regulatory step, rather than directly influencing a specific component of Chl catabolism. Transcriptomic and comparative proteomic profiling further indicated that the nan mutation resulted in the suppressed expression of numerous photosynthesis-related genes and in the induction of genes that are associated with oxidative stress. These data, along with metabolite analyses, suggest that nan fruit employ a number of molecular mechanisms to compensate for the elevated Chl levels and associated photooxidative stress.