Antibacterial and antifungal activity of the three major components found in
Abstract
Essential oils (EO) are volatile compounds produced by the secondary metabolism of aromatic plants. They are complex mixtures whose main components are synthesized by the mevalonic acid and the methyl erythritol phosphate pathways, which lead to the biosynthesis of terpenes, and the shikimic acid pathway, responsible for the biosynthesis of phenylpropanoid compounds. In nature, EOs are stored in the aerial parts of the plant, being of vital importance for their survival due to their antimicrobial properties. In addition, EOs provide protection against herbivores to the aromatic plants and allow them to repel or attract insects because of their strong fragrance, as well as compete with other plants of the same environment. Humans have exploited the properties of their EOs since ancient times, being used as medicinal remedies, among other uses. Currently, aromatic plants are used in pharmaceutical and food industries. One of the most commonly used aromatic plants is thyme. Thyme is a perennial aromatic plant, taxonomically belonging to the genera Thymus and Thymbra, belonging to the family Lamiaceae. These plants are very abundant in the Mediterranean Region. In this review, we focus on the study of the properties and use of EOs of Thymbra capitata (L) Cav. and Thymus hyemalis Lange., whose EOs are rich in phenolic monoterpenes. These compounds are responsible for their antioxidant, anti-inflammatory, anticarcinogenic, antibacterial, antifungal, and antiparasitic properties.
Keywords
- essential oil
- Thymus hyemalis
- Thymbra capitata
- aromatic plant
- antioxidant
- antimicrobial
- carvacrol
- thymol
1. Introduction
Essential oils (EOs) are volatile odorous compounds, are liquids at room temperature, and are produced by aromatic plants, as a result of their secondary metabolism [1, 2].
In nature, the EOs are stored in the secretory cells, cavities, channels, epidermal cells and trichomes of all the aerial organs of the plants, since they are of vital importance for plant survival, due to their antifungal, antibacterial, and antiviral activities. Also, they provide plants protection against herbivores and allow plants to compete with other plants, acting as allelopathic compounds. In addition, they are involved in pollination, attracting insects which favor the dispersal of seeds and pollen [3].
On the other hand, aromatic plants which produce these EOs have been used since ancient times to treat diseases, due to their healing properties. In fact, there are studies that claim that already in Ancient Egypt (2000 BC), these compounds were used as medicinal remedies, beauty products, and in religious rituals. Likewise, Hippocrates (460–377 BC), the father of medicine, studied and documented the properties of 300 aromatic plants, confirming the use of EOs in Ancient Greece. The Romans also showed great interest in the fragrance and properties of the EOs. Dioscorides, a Greek physician and botanist, described in Ancient Rome more than 500 aromatic plants and their EOs, in his book
In the nineteenth century, EO composition was investigated [5]. It is now known that the major components of EOs are synthesized from three biosynthetic pathways: the mevalonic acid pathway, active in the cytosol, and the methyl erythritol phosphate pathway, active in the chloroplast, both of which lead to the biosynthesis of terpenes. A third route is the shikimic acid pathway, responsible for the biosynthesis of phenylpropanoid compounds [6]. EOs of terpene nature are synthesized from isopentenyl pyrophosphate and its isomer dimethylallyl pyrophosphate, which give rise to geranyl diphosphate, precursor of monoterpenes, and farnesyl diphosphate, precursor of sesquiterpenes, as shown in Figure 1. Among the numerous compounds present in the EOs derived from this biosynthetic pathway, thymol and carvacrol (isomers) stand out due to the numerous properties that are granted to them. As seen in Figure 2, both are phenolic monoterpenes synthesized from p-cymene, whose precursor is γ-terpinene [7].
Thymol and carvacrol are usually found in thyme EOs. On the other hand, compounds such as alcohols, aldehydes, ketones, esters, and, less frequently, carboxylic acids, as well as aromatic compounds such as phenolic ethers and aromatic esters are also present, although in a significantly lower proportion than the previous ones [6]. The properties of the EOs are mainly attributed to the major compounds, thymol and carvacrol; however it has been observed that these compounds can interact with the minority compounds, causing synergistic or antagonistic effects, thus influencing the properties of the EO [8]. In addition, the chemical composition varies according to environmental and genetic factors, influencing the phenological stage in which the harvest is made on the quality and quantity of the EO [9, 10]. The high variability occurs even within the same species, there being different major compounds among the specimens of the populations, which gives rise to the existence of different chemotypes [11].
Nowadays, many of the active ingredients used in the development of both traditional and modern drugs are extracted from plant species [12]. For the extraction of the EO, the technique most often used is the hydrodistillation, which consists of submerging fragments of the aromatic plant in boiling water, and so, the volatile compounds are dragged with the vapor, arriving at a condenser which separates them, and thus, the EO is obtained. Other conventional techniques such as steam distillation or extraction with volatile solvents as well as hydrodistillation by microwaves or extraction by supercritical fluids can be used [4]. After extracting, EOs can be analyzed by gas chromatography and mass spectrophotometry (GC–MS). GC allows the separation of the components of a complex mixture from the EO, and the MS serves for the identification of the individual components, already separated [13].
Interest in EOs has skyrocketed in recent times. The demand for “natural” products increases year after year, and aromatic plants and EOs are becoming part of daily life. Likewise, more and more people are investigating the use of compounds obtained from plant extracts in medicine, such is the case of the EOs of many aromatic plants such as lavender (
It is estimated that more than 250,000 hectares are currently used to produce about 250 different plant extracts and, so on, different EOs, so they have a high socioeconomic importance in the places where they are produced, being generally rural areas in developing countries. These EOs are often used in the food industry as well as in other products of daily use such as bath gels, soaps, detergents, oral care products and body lotions. They are also widely used in aromatherapy (International Trade Center 2014). This justifies that, on a global level, 45,000 tons of EOs are produced annually, which implies an investment of more than 600 million euros, according to a study carried out by the Ministry of Agriculture of France. The main exporters are China, the USA, Brazil, EU countries, India, and Indonesia, and the largest imports are Switzerland, the USA, EU countries, Japan, and Canada [16].
In the industry of the EOs, one of the aromatic plants with greater use is thyme. Thyme is a small shrub and perennial aromatic plant, belonging taxonomically to the genera
Within the Region of Murcia, we found several species of thyme, two of them being of special relevance, both for their properties and for their environmental situation [18]: (1)
This work focuses on these two species, due to the fact that
2. Bioactive compounds of T. capitata EO
According to several studies, the EO of
In this sense, the experiments carried out by [10] confirmed the existence of these three chemotypes, which supports the hypothesis that
Finally, in relation to other components found in smaller proportion (such as geraniol, camphor, or β-caryophyllene, among others), there is a high variability between populations and even within the same population [24, 31]. This variability can influence the bioactivity of
3. Bioactive compounds of T. hyemalis EO
In a study conducted by [32], it was observed that
The variability in the chemical composition of
One of the studies that supports the previous statement were carried out by Jordán et al. [36], where it was observed that, in the case of thymol chemotype, the synthesis of this major compound occurred during the flowering/fruit ripening stage. The precursors of thymol, γ-terpinene, and
Finally, similar to the results of
4. Bioactivity of T. capitata EO
4.1 Antioxidant activity
It has been observed that the EO of
This antioxidant capacity has been widely researched in order to prevent lipid oxidation during the storage of vegetable oils for culinary use, such as olive or sunflower. Likewise, Miguel et al. [26] showed that
Another study conducted by Miguel et al. [9] on the lipid oxidation of peanut and sunflower oils showed a low antioxidant activity of
On the other hand, Galego et al. [41] carried out a study on the antioxidant capacity of EOs extracted from
In addition, it was observed that the antioxidant capacity of the EO of
Faleiro et al. [15] also used the TBARS method to observe the effectiveness of the EO of
It should be noted that the antioxidant activity of
4.2 Antibacterial activity
The antibacterial activity of the EO of
This antibacterial activity of
In addition, Delgado-Adámez et al. [30] showed that the EO extracted from both flowers and fruits of
4.3 Antifungal activity
According to Salgueiro et al. [46], the EO of 22 specimens of
These results agree with Palmeira-de-Oliveira et al. [47, 48], whose studies demonstrated that the EO of
In the case of biofilms, when the concentration of the EO doubled the MIC, a decrease of 80% of its metabolism was observed. Antifungal activity of the EO of
Likewise, it has been observed that carvacrol or
On the other hand, Russo et al. [23] observed that the EO of
4.4 Antiparasitic activity
Machado et al. [22] analyzed the antiparasitic activity of EOs rich in phenolic compounds of the species
5. Bioactivity of T. hyemalis EO
5.1 Antioxidant activity
Several studies have shown the antioxidant activity of
On the other hand, Jennan et al. [50] compared the activity of the EO of
5.2 Antibacterial activity
Rota et al. [32] conducted a study on the antimicrobial activity of EOs from several thyme species, specifically,
Some microorganisms, such as
Tepe et al. [51] also investigated the in vitro antimicrobial activity of
Microorganisms | MIC Commercially available essential oil component | ||
---|---|---|---|
Thymol | Carvacrol | p-cymene | |
1.95 | 0.48 | 250.00 | |
0.97 | 0.24 | 250.00 | |
0.97 | 1.95 | 250.00 | |
1.95 | 0.48 | 250.00 | |
1.95 | 3.90 | 250.00 | |
1.95 | 1.95 | 250.00 | |
15.62 | 7.81 | 250.00 | |
0.97 | 0.24 | 15.62 |
This activity of carvacrol was compared with the antibacterial activity of thymol, the second most important component of
5.3 Antifungal activity
Tepe et al. [51] demonstrated the antifungal activity of
6. Discussion
In general, all the data together show that
According to the literature reviewed, the biological activity found in the EOs is clearly related to the chemical composition of them. As regards the
For both species (
Likewise, the moment in which the specimens are harvested influences the composition of EOs, since it varies throughout the life cycle of these plants, affecting their bioactivities. In fact, in
On the other hand, the results show that in the absence of environmental variations, the different chemotypes are genetically determined. This has been observed for the EO of other medicinal plants, such as
Regarding the antioxidant activity, the results indicate that it depends on the concentration. The EO of
This is due, in large part, to the fact that the antioxidant activity of EO (as well as the rest of activities) does not depend only on the majority component but also depends on the synergistic or antagonistic interactions of the majority component with other minority components, which according to the phenological stage will be different [58]. However, in the literature reviewed, some authors indicate the absence of these interactions because isolated carvacrol has been shown to have activity on its own [27, 39]. However, the EO of
On the other hand, both EOs extracted from
Regarding their antibacterial activity, EOs have shown a potent bactericidal effect against a large number of Gram+ and Gram- species. In this sense, EO of
Likewise, both EOs from
In relation to the antifungal activity, it has been demonstrated for the EO of
In addition, the
Currently, the trade and use of thyme EOs is more focused on species such as
Finally, in relation to the mechanism of action by which EOs have their different effects, it is not clear. It is known that all the activities mentioned are dependent on the concentration at which they are used. As we have seen throughout this work, the results vary depending on the dose of EO used. With regard to the antifungal and antibacterial activity, the EO acts by affecting the membrane permeability of the pathogen. At high concentrations, the EO denatures the proteins, whereas if the concentration is low, the enzymatic activity related to the production of energy is affected.
It has also been observed that, in the case of
7. Conclusion
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