Chemical composition and evaluation of antioxidant, antimicrobial and antiproliferative activities of Tuber and Terfezia truffles

doi:10.1016/j.foodres.2020.110071

Food Research International

Volume 140, February 2021, 110071

https://doi.org/10.1016/j.foodres.2020.110071 Get rights and content

Highlights

•
Truffles are a rich source of carbohydrates and proteins.
•
Oleic, linoleic and palmitic acids were the main truffle fatty acids.
•
Oxalic and fumaric acids and tocopherols (α, β, γ and δ) were found in truffles.
•
T. magnatum contained the highest TPC values and the highest antioxidant activity.
•
Terfezia and T. gennadii extracts revealed antiproliferative activity.

Abstract

Ten truffle species of Tuber and Terfezia genera were chemical characterized, assessing their proximate composition, individual nutrient compounds and some bioactive molecules. The bioactive properties of these species were also evaluated, namely their antioxidant, antimicrobial and cytotoxic potential. Carbohydrates were the main macronutrients present in truffles, followed by proteins. Furthermore, the levels of polyunsaturated fatty acids (PUFA), subsequently presented as a percentage, were higher in truffles (38.2–79.3%) except in Tuber magnatum and Terfezia arenaria, which have a more saturated fatty acids (SFA) profile (70.7% and 53.7%, respectively). Comparing the species, T. magnatum revealed the highest levels of total phenolic compounds (TPC) (290 mg GAE/100 g truffle), as also the best results in the four methods used to evaluate the antioxidant activity. On the other hand, only five extracts obtained from some studied truffle species (Terfezia magnusii, Tuber aestivum, Tuber gennadii, and Tuber melanosporum) showed a slight inhibition of microbial growth, tested against different bacteria. Terfezia and T. gennadii extracts, showed potential to inhibit the cellular growth of NCI-H460, HeLa, HepG2, and MCF-7 cell lines (GI₅₀ concentrations range: 19–78, 33–301, 83–321 and 102–321 µg/mL, respectively), indicating anti-proliferative activity. Nevertheless, T. arenaria revealed some potential hepatotoxicity, inhibiting the growth of PLP2 cells (GI₅₀ concentration of 220 µg/mL), a primary cell culture obtained from porcine liver.

Graphical abstract

Introduction

Edible fungi can be classified in two groups, epigeous carpophores commonly known as mushrooms, and hypogeous species that grow underground, known as truffles (Lee et al., 2020). The most economically important genus of truffles is the genus Tuber, which includes the species T. magnatum Pico (white truffle), Tuber melanosporum Vittad. (black truffle), or Tuber aestivum Vittad. (summer truffle), highly appreciated for their distinctive aroma (Costa et al., 2015, Culleré et al., 2013). These species are traditionally harvested in Italy, France, and Spain. Outside Europe, whereas Tuber indicum Cooke & Massee is exclusively harvested in Asia (Reyna & Garcia-Barreda, 2014). Other genera, such as Terfezia or Tirmania, have culinary interest, but their organoleptic qualities are less appreciated worldwide. These genera, also called desert truffles, are mostly endemic to the arid and semi-arid areas of the Mediterranean basin (Zambonelli et al., 2014).

The investigation of their nutritional profile showed that fresh mushrooms, including truffles, are a rich source of carbohydrates and proteins (Kalač, 2013). Some minor compounds such as minerals, amino acids, and fatty acids were generally determined in truffles (Lee et al., 2020). However, other minor chemical constituents, specifically phenolic compounds or tocopherols, only have been studied in T. aestivum and T. magnatum truffles (Beara et al., 2014, Shah et al., 2020). Some of these compounds, even in synergy with others, have been reported to have biological activity, namely free radical scavenging activity, metal chelation, or inhibition of lipid oxidation against reactive oxygen species (ROS) (Sánchez, 2017). Furthermore, some reports on bioactivities (antiviral, antimicrobial, antimutagenic, antioxidant and anti-inflammatory) of desert truffles belonging to Terfezia and Tirmania spp. have been investigated (Dahham et al., 2018, Stojković et al., 2013, Vahdani et al., 2017).

As far as we know, the biological activity of Tuber genus was scarcely explored in three species. There are studies reporting that the aqueous and ethanolic extracts from T. aestivum exhibit an antimutagenic effect (Fratianni, Luccia, Coppola, & Nazzaro, 2007), while its methanolic extract inhibited ABAP (2′-azobis-(2-amidinopropane)-dihydrochloride)-induced lipid peroxidation (Villares, García-Lafuente, Guillamón, & Ramos, 2012). Also, T. magnatum showed antioxidant, anti-inflammatory and cytotoxic activities (Beara et al., 2014) and T. indicum showed antioxidant activity (Guo, Wei, Sun, Hou, & Fan, 2011). Recently, we reported anti-immunomodulatory properties in T. melanosporum (Tejedor-Calvo, Morales, García-Barreda, et al., 2020). Besides, several studies have demonstrated the antimicrobial activity of different extracts of several epigeous carpophores (Venturini, Rivera, Gonzalez, & Blanco, 2008). Regarding hypogeous fungi, the methanolic and ethanolic extracts of Terfezia truffles showed antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, and Pseudomonas aeruginosa (Janakat, Al-Fakhiri, & Sallal, 2004). Within this genus, the species T. arenaria (Moris) Trappe showed antimicrobial activity against Enterococcus faecalis and Escherichia coli, among other pathogenic bacteria, yeast and filamentous fungi (Harir et al., 2019). Nevertheless, to the best of our knowledge, there are no reports on the biological activities of Tuber brumale Vittad., T. gennadii (Chatin) Pat., Tuber oligospermum (Tul. & C. Tul.) Trappe, Terfezia leptoderma Tul. & C. Tul., and Terfezia magnusii Mattir.

Consequently, this study aimed to perform for first time a comprehensive investigation of the proximate analysis, chemical composition (i.e. fatty acids, organic and phenolic acids, proteins, tocopherols, and soluble sugars), as well as antioxidant, antimicrobial and anti-proliferative bioactivities of ten truffles species belonging to Tuber and Terfezia genera.

Section snippets

Truffle samples

Fresh hypogeous edible carpophores belonging to ten species were studied, seven of the genus Tuber (T. aestivum, T. brumale, T. indicum, T. gennadii, T. magnatum, T. melanosporum, and T. oligospermum), and three of the genus Terfezia (T. arenaria, T. leptoderma, T. magnusii). T. aestivum, T. brumale, and T. melanosporum were directly harvested from the orchard. The rest was obtained from natural truffle grounds, except T. indicum and T. magnatum that were supplied by Espora Gourmet Company. The

Proximate analysis

The results of the proximate analysis and estimated energetic value obtained for the ten truffle species are shown in Table 2. Protein was found in high levels and varied between 14.04 and 24.15 g/100 g dw, the highest levels registered by T. magnatum and T. arenaria. This late species also revealed the highest amounts of crude fat (5.10 g/100 g dw). However, the fat content was relatively low for all species, ranging between 0.99 and 5.10 g/100 g dw. Ash content varied between approximately

Conclusions

Tuber and Terfezia truffles highlighted by their PUFA, β-tocopherols and phenolic acids profile and contents especially T. brumale and T. melanosporum for the first two compounds, and T. magnatum for the phenol content. However, in respect to the anti/proliferative effects against human tumor cell all Terfezia species revealed better effects than Tuber ones. Thus, it was not possible to select a particular compound or group of compounds as responsible for the species' bioactivity that could be

CRediT authorship contribution statement

Eva Tejedor-Calvo: Investigation, Writing - original draft. Khira Amara: Investigation, Writing - original draft. Filipa S. Reis: Investigation, Methodology, Writing - original draft. Lillian Barros: Conceptualization, Methodology, Data curation, Writing - review & editing. Anabela Martins: Supervision, Writing - review & editing. Ricardo C. Calhelha: Investigation, Methodology. Maria Eugenia Venturini: Supervision, Project administration. Domingo Blanco: Funding acquisition, Supervision. Diego

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

The authors thank the National Institute for Agricultural and Food Research and Technology (project RTA2010-00070-C02-02) for financial support; to Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); and to national funding by FCT, PI, through the institutional scientific employment program-contract for L. Barros’s contract; to the project Valor Natural for the contract of F. Reis (Mobilized Project

References (51)

A. Al-Laith
Antioxidant components and antioxidant/antiradical activities of desert truffle (Tirmania nivea) from various Middle Eastern origins
Journal of Food Composition and Analysis
(2010)
L. Barros et al.
Bioactivity and chemical characterization in hydrophilic and lipophilic compounds of Chenopodium ambrosioides L
Journal of Functional Foods
(2013)
I.N. Beara et al.
Phenolic profile, antioxidant, anti-inflammatory and cytotoxic activities of black (Tuber aestivum Vittad.) and white (Tuber magnatum Pico) truffles
Food Chemistry
(2014)
M. Carocho et al.
A review on antioxidants, prooxidants and related controversy: Natural and synthetic compounds, screening and analysis methodologies and future perspectives
Food and Chemical Toxicology
(2013)
R. Costa et al.
Screening of volatile compounds composition of white truffle during storage by GCxGC-(FID/MS) and gas sensor array analyses
LWT - Food Science and Technology
(2015)
L. Culleré et al.
Potential aromatic compounds as markers to differentiate between Tuber melanosporum and Tuber indicum truffles
Food Chemistry
(2013)
S.S. Dahham et al.
Antioxidant, anticancer, apoptosis properties and chemical composition of black truffle Terfezia claveryi
Saudi Journal of Biological Sciences
(2018)
M.I. Dias et al.
Nutritional parameters of infusions and decoctions obtained from Fragaria vesca L. roots and vegetative parts
LWT - Food Science and Technology
(2015)
A. Elaissi et al.
Antibacterial activity and chemical composition of 20 Eucalyptus species’ essential oils
Food Chemistry
(2011)
I.C.F.R. Ferreira et al.
Free-radical scavenging capacity and reducing power of wild edible mushrooms from northeast Portugal: Individual cap and stipe activity
Food Chemistry
(2007)

Cited by (22)

White Acqualagna truffle (Tuber magnatum Pico): Evaluation of volatile and non-volatile profiles by GC-MS, sensory analyses and elemental composition by ICP-MS
2024, Food Chemistry
The White Truffle is the most expensive edible underground mushroom. In this study the first characterization of the Acqualagna white truffle was delivered, taking into consideration the soil of origin and the human perception. The volatile profile was identified by GC–MS and compared with the descriptors obtained by sensory analysis. The non-volatile characterization was done using elemental composition by ICP-MS analysis, elemental analysis, and spectrophotometric assays. The volatile profile consists mainly of bis(methylthio)methane (78.72%) and other minor constituents, linked to seven odorant descriptors: garlic-like, nutty-like, geosmine-like, floral, mushroom-like, pungent and green/herbal. ICP-MS revealed that truffle has a higher content of K, P, S, Ca and Mg (97% of the elements investigated) and that it assimilates the Rare Earth Elements (REE) from the soil without discriminating them. In conclusion, this project is the first step for the enhancement of local food, linked to the territory conditions in which it is produced.
Extraction and trapping of truffle flavoring compounds into food matrices using supercritical CO<inf>2</inf>
2023, Food Research International
A supercritical fluid extraction methodology was used to extract flavoring and bioactive compounds from truffles. Some parameters such as CO₂ flow rate (1–3 mg/mL), extraction time (15–90 min) and different trapping food matrices (grape seed oil, gelatin, agar agar and water) were optimized using response surface methodology to enhance extraction and trapping yields. The optimal conditions (2.27 mg/mL CO₂ flow rate, 82.5 min when using 40 °C and 30 MPa, with 1 mL grape seed oil as trapping matrix) obtained with Tuber melanosporum were applied to three different truffle species: Terfezia claveryi, Tuber aestivum and Tuber indicum. A total of 32 metabolites were profiled in the extracts using ultra-high-performance supercritical fluid chromatography coupled to quadrupole time-of-flight mass spectrometry. Compounds such as brassicasterol ergosta-7,22-dienol, oleic and linoleic acid were found at similar amounts in all the extracts but other molecules (e.g. fungal sterols) showed a particular distribution depending on the specie studied and whether a trapping matrix was used at the SFE outlet.
Isolation and identification of compounds from truffle Reddellomyces westraliensis and their antioxidant, cytotoxic and enzyme inhibitory activities
2022, Process Biochemistry
Citation Excerpt :
TPC of the acetone (54.95 ± 9.50–79.09 ± 9.71 mg/mL) and methanol (21.84 ± 4.18–50.51 ± 4.40 mg/mL) extracts of Terfezia claveryi, Terfezia boudieri, Terfezia olbiensis, Picoa lefebvrei, and Picoa juniper were calculated by Fidan et al. [17]. The level of TPC was found in the range of 22 ± 10–290 ± 5 in the methanol extracts of Tuber aestivum, Tuber brumale, Tuber indicum, Tuber gennadii, Tuber magnatum, Tuber melanosporum, Tuber oligospermum, Terfezia arenaria, Terfezia leptoderma and Terfezia magnussi [18]. TPC of R. westraliensis extracts were correlated with TPC results of different truffle species.
Truffles have become one of the world’s most important mushrooms owing to their distinctive features. To the best of our knowledge, this is the first study on Reddellomyces westraliensis in terms of isolation and identification of compounds and screening versatile bioactivities. Chromatographic studies on R. westraliensis enabled to isolate brassicasterol (1), ergosta-7,9,22-trien-3β-ol (2), ergosterol peroxide (3), adenosine (4), D-ribitol (5), fumaric acid (6) and mannitol (7). The structures of the compounds were verified by IR, NMR, MS techniques. Antioxidant, cytotoxic and cholinesterase, tyrosinase, urease, α-amylase and α-glucosidase inhibitory activities were assessed. The methanol was the suitable extraction solvent high content of phenolic compound (18.94 ± 0.18 μg PEs/mg extract). Compound 4 showed higher DPPH^• (IC₅₀: 8.38 ± 0.40 µg/mL) and ABTS^•+ (IC₅₀: 6.74 ± 0.22 µg/mL) scavenging activity than α-tocopherol and BHA. Compound 3 was observed as a potent inhibitor against AChE (34.03 ± 0.13 %), α-amylase (34.28 ± 0.36 %) and BChE (66.31 ± 0.08 %). Compounds 1 (IC₅₀: 5.85 ± 0.16 µg/mL) and 3 (IC₅₀: 6.98 ± 0.24 µg/mL) indicated higher urease inhibitory activity than thiourea (IC₅₀: 7.87 ± 0.18 µg/mL). Compounds 4 (9.61 ± 0.92 %) and 7 (21.56 ± 0.74 %) indicated higher α-glucosidase and tyrosinase inhibitory activity, respectively. Compound 2 exhibited the best cytotoxicity against H1299 (IC₅₀: 38.62 ± 0.46 µg/mL) and MCF-7 (IC₅₀: 24.93 ± 0.80 µg/mL). HPLC-DAD analysis exhibited the existence of fumaric acid (27.20 ± 0.06 µg/g) as the major compound. This study revealed that R. westraliensis can be approved as antioxidant, cytotoxic, and enzyme inhibitor sources in conjunction with its varied bioactive compounds.
Eco-archaeological excavation techniques reveal snapshots of subterranean truffle growth
2021, Fungal Biology
Citation Excerpt :
The fruiting-bearing seasons of most truffle species are in winter. For instance, the Périgord black truffle is usually harvested between November and March, whereas the white Piedmont truffle (Tuber magnatum) is mainly harvested from late summer to winter (Callot, 1999; Tejedor-Calvo et al., 2021), and rarely after January. The Burgundy truffle, however, can produce fruitbodies during a much longer season from at least June until March of the following year (see Čejka et al., 2020 for a review of the ecological requirements of Burgundy and Périgord truffles).
Despite its status as a highly-prized and coveted fungi in gastronomy, many aspects of the subterranean life cycle of the Burgundy truffle (Tuber aestivum) are still unknown, because in situ observations of the formation and maturation of truffle fruitbodies remain difficult. Here, we adopted a suite of archaeological fine-scale excavating techniques to provide unique spatiotemporal snapshots of Burgundy truffle growth at three sites in southern Germany. We also recorded the relative position, fresh weight, maturity level and genotype composition of all excavated fruitbodies. Varying by a factor of thousand, the fresh weight of 73 truffle ranged from 0.1 to 103.2 g, with individual maturity levels likely representing different life cycle stages from completely unripe to fully ripe and even decaying. While only a slightly positive relationship between fruitbody weight and maturity level was found, our results suggest that genetically distinct specimens can exhibit different life cycle stages at the same period of time and under the same environmental conditions. We therefore argue that truffles are likely able to grow, mature and ripe simultaneously between early summer and late winter of the following year. Our case study should encourage further eco-archaeological truffle excavations under different biogeographic settings and at different seasons of the year to gain deeper insights into the fungi's subterranean ecology. The expected cross-disciplinary findings will help truffle hunters and farmers to improve their harvest practices and management strategies.
Vitamin E as an essential micronutrient for human health: Common, novel, and unexplored dietary sources
2021, Free Radical Biology and Medicine
Citation Excerpt :
Other—perhaps less obvious—sources of vitamin E should also be considered. Tejedor-Calvo et al. [113] investigated the tocopherol composition of edible fungi (Tuber and Terfezia truffles from China, Spain, and Italy). Among the ten species analyzed, T. brumale, T. magnatum, T. melanosporum, and T. oligospermum had the highest tocopherol content (59.7, 37.8, 32.6, and 27.8 mg/100 g dw, respectively).
Vitamin E comprises a group of vitamers that includes tocopherols and tocotrienols. They occur in four homologues according to the number and position of methyl groups attached to the chromanol ring. Vitamin E, a liposoluble antioxidant, may participate as an adjuvant in the prevention and treatment of cardiovascular, neurological, and aging-related diseases. Furthermore, vitamin E has applications in the food industry as a natural additive. In this contribution, the most recent information on the dietary sources of vitamin E, including common, novel, and unexplored sources, is presented. Common edible oils, such as those of corn, olive, palm, rice bran, and peanut, represent the most prominent sources of vitamin E. However, specialty and underutilized oils such as those obtained from tree nuts, fruit seeds, and by-products, emerge as novel sources of this important micronutrient. Complementary studies should examine the tocotrienol content of vitamin E dietary sources to better understand the different biological functions of these vitamers.
Supercritical CO<inf>2</inf> extraction method of aromatic compounds from truffles
2021, LWT
Citation Excerpt :
Therefore, this technique could be a good proceeding to extract aromatic compounds from truffles. Truffles contain other valuable compounds i.e. β-glucans, or specific fugal sterols (Tejedor-Calvo et al., 2019; Tejedor-Calvo, Amara, et al., 2020) with interesting biological activities such as immunomodulatory and hypocholesterolemic properties (Patel, Rauf, Khan, Khalid, & Mubarak, 2017). These molecules could remain in the residual cake after the extraction of the aromatic compounds as a byproduct, and they could be also extracted to design novel functional foods.
Truffles are a well-known worldwide product mainly appreciated by their unique aroma, which is composed by more than 50 volatile compounds. However, to this day, no one has accomplished to find the aromatic key that evokes the real aroma of truffles for its use as food flavoring. Among them, black truffle was selected for extraction with supercritical fluids using CO₂ as solvent recovering natural truffle aroma fraction. To achieve the optimal extraction ratio, time, pressure and grapeseed oil addition to the separators were evaluated. Aroma from black truffle powder, extracts obtained, and residual cakes fractions were characterized by headspace gas chromatography-spectrometry and olfactometry techniques. The results indicated that optimal extraction conditions were 30 MPa for 3 h. Also, grapeseed oil addition enhanced trapping some key truffle aromatic compounds as 2,3-butanodione, 2-methyl-1-butanol, octanal and dimethyl disulphide. Olfactometry study showed the aromatic profile of the extracts indicating the molecules ethyl pentanoate (fruity), 1-hexen-3-one (metallic) and ethyl hexanoate (fruity) as the main compounds of extracts samples. For the first time, a natural truffle aroma has been obtained using low-value truffles. After aromatic extraction, carbohydrates, proteins, and phenolic compounds were analysed within the residues, showing a potential source of bioactive compounds.

View all citing articles on Scopus

View full text

Chemical composition and evaluation of antioxidant, antimicrobial and antiproliferative activities of Tuber and Terfezia truffles

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Truffle samples

Proximate analysis

Conclusions

CRediT authorship contribution statement

Declaration of Competing Interest

Acknowledgements

Journal of Food Composition and Analysis

Journal of Functional Foods

Food Chemistry

Food and Chemical Toxicology

LWT - Food Science and Technology

Food Chemistry

Saudi Journal of Biological Sciences

LWT - Food Science and Technology

Food Chemistry

Food Chemistry

Food Chemistry

Food Chemistry

Food Chemistry

Food Chemistry

Food Chemistry

Food and Chemical Toxicology

Food Microbiology

Nutrition Research

Synthetic and Systems Biotechnology

Food Research International

Food Research International

Postharvest Biology and Technology

Journal of Food Composition and Analysis

Journal of Food Composition and Analysis

Food Research International