Gigliola Borgonovo

Taste-guided identification of high potency TRPA1 agonists from Perilla frutescens

Perilla frutescens is a food plant widely used in Asian cuisine. This plant was investigated for its interesting taste and somatosensory properties. Perillaldehyde and perillaketone are among the components of the aromatic extracts from P. Frutescens. These compounds were shown here to activate the cloned TRPA1 channel when expressed in an heterologous cell system and are therefore suggested to be responsible for the chemesthetic properties of this plant.

The taste of D- and L-amino acids: In vitro binding assays with cloned human bitter (TAS2Rs) and sweet (TAS1R2/TAS1R3) receptors

The taste of different enantiomeric forms of amino acids has been deeply investigated because it represents the most impressive case of correlation between stereochemistry and flavour. Herein, we aimed to elucidate the molecular activity of d- and l-amino acids using an in vitro system based on a cellular model overexpressing sweet and bitter receptors, and to analyse the correlation between in vitro and sensory studies. With our work we demonstrated specific enantiomeric activities for several amino acids on TAS1R2-TAS1R3 sweet receptor. Moreover, we proved interaction of tryptophan and phenylalanine with a specific group of TAS2Rs bitter receptors, confirming and improving the results recently obtained in the tasting of amino acids. In addition, we provide the first systematic analysis of l- and d-amino acid actions on the sweet heterodimeric receptor.

Analogues of perillaketone as highly potent agonists of TRPA1 channel.

Transient receptor potential (TRP) channels represent interesting molecular target structures involved in a number of different physiological and pathophysiological systems. In particular, TRPA1 channel is involved in nociception and in sensory perception of many pungent chemesthetic compounds, which are widespread in spices and food plants, including Perilla frutescens. A natural compound from P. frutescens (isoegomaketone) and 16 synthetic derivatives of perillaketone have been prepared and tested in vitro on rTRPA1 expressed in HEK293 cells and their potency, efficacy and desensibilisation activity measured. Most derivatives proved to be high potency agonists of TRPA1, with a potency higher than most natural agonists reported in the literature. These furylketones derivatives, represent a new class of chemical structures active on TRPA1 with many potential applications in the agrifood and pharmaceutical industry.

Taste-Active Compounds in a Traditional Italian Food: ‘Lampascioni’

Nature is a rich source of taste‐active compounds, in particular of plant origin, many of which have unusual tastes. Many of these are found in traditional food, where spontaneous plants are used as ingredients. Some taste‐active compounds were identified in the bulbs of Muscari comosum, a spontaneous plant belonging to the family of the Liliaceae, very common in the Mediterranean area, and used in traditional gastronomy (called ‘lampascioni’ in South Italy). The bulbs were extracted with a series of solvents of different polarity. The different fractions were submitted to a preliminary sensory evaluation, and the most interesting ones, characterized by a strong bitter taste and some chemestetic properties, were submitted to further purification and structural analysis. From the ethereal extract, several 3‐benzyl‐4‐chromanones and one stilbene derivative were isolated. Pure compounds were examined for their taste activity by means of sensory evaluation, and proved to be responsible for the characteristic taste of this food. Some of these compounds have been synthesized de novo to confirm their structure.

Synthesis and sweet taste of optically active (–)-haematoxylin and of some (±)-haematoxylin derivatives

In order to explain the sweet taste of the natural polyphenolic compound (+)-haematoxylin 1, four (±)-haematoxylin derivatives 4–7 and the enantiomer (–)-haematoxylin have been synthesized and tasted. Unlike haematoxylin, the derivatives 4–7 have a restricted number of different possibilities of binding to the sweet taste receptor according to the Shallenberger–Acree–Kier model. This allowed the study of the most likely orientation of these compounds in the active site of receptor. The results are supported by the comparison of the molecular structures with the receptor models of Temussi–Goodman and Tinti–Nofre. The synthesis of the (–)-enantiomer of haematoxylin also allows for discussion of the relationship between configuration and sweet taste in these compounds.

Phytochemicals from Ruta graveolens Activate TAS2R Bitter Taste Receptors and TRP Channels Involved in Gustation and Nociception

Ruta graveolens (rue) is a spontaneous plant in the Mediterranean area with a strong aroma and a very intense bitter taste, used in gastronomy and in folk medicine. From the leaves, stems and fruits of rue, we isolated rutin, rutamarin, three furanocoumarins, two quinolinic alkaloids, a dicoumarin and two long chain ketones. Bitter taste and chemesthetic properties have been evaluated by in vitro assays with twenty receptors of the TAS2R family and four TRP ion channels involved in gustation and nociception. Among the alkaloids, skimmianine was active as a specific agonist of T2R14, whereas kokusaginin did not activate any of the tested receptors. The furanocoumarins activates TAS2R10, 14, and 49 with different degrees of selectivity, as well as the TRPA1 somatosensory ion channel. Rutamarin is an agonist of TRPM5 and TRPV1 and a strong antagonist of TRPM8 ion channels.

Bitter or not? BitterPredict, a tool for predicting taste from chemical structure

Bitter taste is an innately aversive taste modality that is considered to protect animals from consuming toxic compounds. Yet, bitterness is not always noxious and some bitter compounds have beneficial effects on health. Hundreds of bitter compounds were reported (and are accessible via the BitterDB http://bitterdb.agri.huji.ac.il/dbbitter.php), but numerous additional bitter molecules are still unknown. The dramatic chemical diversity of bitterants makes bitterness prediction a difficult task. Here we present a machine learning classifier, BitterPredict, which predicts whether a compound is bitter or not, based on its chemical structure. BitterDB was used as the positive set, and non-bitter molecules were gathered from literature to create the negative set. Adaptive Boosting (AdaBoost), based on decision trees machine-learning algorithm was applied to molecules that were represented using physicochemical and ADME/Tox descriptors. BitterPredict correctly classifies over 80% of the compounds in the hold-out test set, and 70–90% of the compounds in three independent external sets and in sensory test validation, providing a quick and reliable tool for classifying large sets of compounds into bitter and non-bitter groups. BitterPredict suggests that about 40% of random molecules, and a large portion (66%) of clinical and experimental drugs, and of natural products (77%) are bitter.

Response of the European grapevine moth Lobesia botrana to somatosensory-active volatiles emitted by the non-host plant Perilla frutescens

The European grapevine moth Lobesia botrana Denis & Schiffermüller (Lepidoptera: Tortricidae) is a major pest on grapes worldwide. Attempts to develop control methods for this pest based on grape kairomones demonstrate limited success and studies indicate that a major limiting factor is overlap between synthetic kairomones and background odours in the vineyard. Behaviourally active compounds from non‐host plants may thus represent an effective alternative for monitoring and control methods. Extracts from food plants (i.e. from capsicum, garlic and peppermint, which elicit the so‐called somatosensory sensation) are traditionally used in agriculture for the control of pest insects. Among those plants, Perilla frutescens L. (Lamiales: Lamiaceae), native of Asia, contains compounds activating sensory ion channels in mammals, which are known to be involved in the perception of somatosensory compounds and are expressed in tortricid moth antennae. In the present study, in search of non‐host volatiles with potential application in pest control, essential oil metabolites isolated from P. frutescens are screened for biological activity on the olfactory system of L. botrana. The compounds (S)‐(−)‐perillaldehyde and isoegomaketone, which are released from different P. frutescens varieties, are identified by gas chromatography‐coupled electroantennographic detection. In a dual‐choice oviposition test, females show a preference for a combination of host odours and perillaldehyde, preferring this over a host‐plant odour bouquet alone. In Y‐tube olfactometer assays, virgin males show a higher level of activity in the presence of isoegomaketone, even if not significantly responsive to the compound.

Sub-lethal concentrations of Muscari comosum bulb extract suppress adhesion and induce detachment of sessile yeast cells

The formation of yeast biofilm on food industry equipment can lead to serious hygiene problems and economic losses due to food spoilage and equipment impairment. This study explored the ability of a sub-lethal concentration of the bulb extract of Muscari comosum to modulate adhesion of Candida albicans and subsequent biofilm development by this fungus. The HPLC profile of the ethanolic bulb extract showed phenolic constituents, which were found to undergo Folin-Ciocalteu reagent reduction. Prior to the adhesion tests, it was shown that up to 4000 mg l−1 of natural extract did not adversely affect fungal growth nor did it act as a carbon energy source for C. albicans. Mathematical models predicted that 4000 mg l−1 and 700 mg l−1 of bulb extract would cause more than 98% reduction in fungal coverage on abiotic surfaces, without killing the planktonic cells. When added to C. albicans biofilm, the natural extract was shown to induce the dispersion of sessile cells in a dose-dependent manner.

Sensory and hedonic perceptions of italian and korean subjects: a Cross-Cultural study of Perilla Frutescens

Differences in perceptions of perilla by Korean and Italian subjects were analyzed. Descriptive analysis was used to verify how Italian assessors described the sensory properties of an unknown product. Italian subjects used the attributes grass, fennel, balsamic, and almond to describe the unfamiliar odor of perilla and perceived perilla as being bitter, astringent and as having a cooling and tingling effect. Projective mapping and hedonic testing were used to compare Italian and Korean subjects odor perceptions and acceptability. Napping results showed that both groups had similar odor perceptions, but Italian subjects tended to associate perilla with other odors whereas Korean subjects were able to specifically identify perilla. Also, Korean subjects generally gave low scores for perilla beverages, but not as low as Italian subjects, suggesting that familiarity plays an important role in food appreciation.