Diallyl Trisulfide, the Antifungal Component of Garlic Essential Oil and the Bioactivity of Its Nanoemulsions Formed by Spontaneous Emulsification

The aim of this study was to evaluate the chemical compounds of garlic essential oil (EO), and determine the antifungal efficacy of garlic EO and its major components, diallyl trisulfide and its nanoemulsions against wood-rotting fungi, Trametes hirsuta and Laetiporus sulphureus. GC-MS analysis revealed that the major constituents of garlic EO were diallyl trisulfide (39.79%), diallyl disulfide (32.91%), and diallyl sulfide (7.02%). In antifungal activity, the IC50 value of garlic EO against T. hirsuta and L. sulphureus were 137.3 and 44.6 μg/mL, respectively. Results from the antifungal tests demonstrated that the three major constituents were shown to have good antifungal activity, in which, diallyl trisulfide was the most effective against T. hirsuta and L. sulphureus, with the IC50 values of 56.1 and 31.6 μg/mL, respectively. The diallyl trisulfide nanoemulsions showed high antifungal efficacy against the examined wood-rotting fungi, and as the amount of diallyl trisulfide in the lipid phase increases, the antifungal efficacy of the nanoemulsions increases. These results showed that the nanoemulsions and normal emulsion of diallyl trisulfide have potential to develop into a natural wood preservative.

Investigation of promoter regions, motifs, and CpG islands in the regulation of gene expression in Trametes hirsuta strain 072

Background In silico analysis of transcription start sites, promoter regions, transcription factors and their binding sites, and CpG islands for the Trametes hirsuta strain 072 genome were performed to understand the regulation mechanisms of gene expression and its genetic variations in the genomes. Therefore, a computational survey was carried out for the Trametes hirsuta strain 072 genome with the open reading frames from the National Center for Biotechnology Information database. Seventeen functional sequences were used to analyze promoter regions and their regulatory elements. Result The present study revealed that 94% of Trametes hirsuta strain 072 genes contained more than two TSSs. Among these identified TSSs, a TSS with the highest predictive score was considered to determine a promoter region of the genes. Moreover, a total of five common candidate motifs such as MotI, MotII, MotIII, MotIV, and MotV were identified. Among these motifs, motif IV was investigated as the common promoter motif for 41.17% of genes that serve as binding sites for transcription factors (TFs) involved in the expression regulation of Trametes hirsuta strain 072 genes. Motif IV was also compared to registered motifs in publically available databases to see if they are similar to known regulatory motifs for TF using TOMTOM web server. Hence, it was revealed that MotIV might serve as the binding site mainly for the leucine zipper TF gene family to regulate a gene expression of Trametes hirsuta strain 072. Regarding CpG island determination, it was concluded that there is no CpG island in both promoter and gene body regions of the Trametes hirsuta strain 072 genome. Conclusions This study provides a better insight into further molecular characterization which aimed to efficiently exploit a white rot fungus, Trametes hirsuta strain 072, for several biotechnological applications aimed to revitalize a severely contaminated environment.

Potential use of basidiomycota Trametes hirsuta MT-17.24 in biodegradation of polyanionic cellulose

Despite their efficiency, existing methods to dispose of drilling fluids used in the construction of oil and gas wells (chemical treatment of spent solutions, thermal method, thickening) are often expensive and unsustainable. Basidiomycota are natural xylotroph destructors that process lignocellulosic substrate – one of the most stable biopolymers in nature. Prospects for using enzyme preparations based on Basidiomycota as biodestructors of organic substances are evident due to the high efficiency and zero-waste production. The aim was to obtain an enzyme preparation based on the Trametes hirsute MT-17.24 Basidiomycota strain and evaluate its ability to biodegrade polyanionic cellulose, used as a viscosifier for drilling fluids in the construction and repair of oil and gas wells. Screening of cellulase activity of the following strains was carried out: Fomitopsis pinicola MT-5.21, Fomes fomentarius MT-4.05, Lactarius necator, Schizophyllum commune MT-33.01, Trametes versicolor It-1, Trametes hirsute MT-17.24, Trametes hirsuta MT-24.24. To obtain the enzyme preparation, the T. hirsuta MT-17.24 strain was selected, which demonstrated the highest coefficient of cellulase activity (10.9). A medium for solid-phase cultivation of this strain was selected. Enzymatic activity of the enzyme preparation was studied on a model drilling fluid. A 10-hour experiment showed that the use of a 1% enzyme preparation leads to a decrease in the plastic viscosity of the drilling fluid from 16 to 8 mPa·s. The research results demonstrate the efficiency of enzyme preparations based on Basidiomycota in the biodestruction of polyanionic cellulose.

Elucidation of ligninolysis mechanism of a newly isolated white-rot basidiomycete Trametes hirsuta X-13

Background Lignin is a complex aromatic heteropolymer comprising 15–30% dry weight of the lignocellulose. The complex structural characteristic of lignin renders it difficult for value-added utilization. Exploring efficient lignin-degrading microorganisms and investigating their lignin-degradation mechanisms would be beneficial for promoting lignin valorization. In this study, a newly isolated white-rot basidiomycete, Trametes hirsuta X-13, with capacity to utilize alkaline lignin as the sole substrate was investigated. Results The analysis of the fermentation properties of T. hirsuta X-13 using alkaline lignin as the sole substrate, including the mycelial growth, activities of ligninolytic enzymes and the rates of lignin degradation and decolorization confirmed its great ligninolysis capacity. The maximum lignin degradation rate reached 39.8% after 11 days of T. hirsuta X-13 treatment, which was higher than that of reported fungi under the same condition. Fourier transform infrared spectrometry (FTIR), gas chromatography–mass spectrometry (GC–MS) scanning electron micrographs (SEM), two-dimensional heteronuclear single quantum coherence NMR analysis (2D-HSQC NMR) collaborated with pyrolysis gas chromatography–mass spectrometry (py-GC/MS) analyses proved that lignin structure was severely deconstructed along with amounts of monomer aromatics generated. Furthermore, according to those chemical analysis, in addition to canonical Cα–Cβ breakage, the cleavage of lignin interunit linkages of β–β might also occur by T. hirsuta X-13. Conclusions This study characterized a newly isolated white-rot basidiomycete T. hirsuta X-13 with impressive alkaline lignin degradation ability and provided mechanistic insight into its ligninolysis mechanism, which will be valuable for the development of lignin valorization strategies.

Biodecolorization of Remazol Brilliant Blue–R dye by Tropical White-Rot Fungi and Their Enzymes in The Presence of Guaiacol

The ability of the tropical white-rot fungi and their enzyme to decolorize synthetic dyes was investigated. Production of lignin-modifying enzymes (LMEs) from the three new isolated fungi, namely Trametes hirsuta D7, Ceriporia sp. BIOM 3, and Cymatoderma dendriticum WM01 were observed for 9 days incubation under static condition. The results showed that the LMEs production enhanced in the present of guaiacol. T. hirsuta D7 produced only laccase (Lac), with the highest activity was 22.6 U/L on the 5th-day of the cultivation. At the same time, Ceriporia sp. BIOM 3 and C. dendriticum WM01 secreted both laccases (Lac) with the activities 0.2 U/L and 1.0 U/L, respectively, and manganese peroxidase (MnP) with the activities 0.1 U/L and 1.0 U/L, respectively. Among the fungi, T. hirsuta D7 efficiently degraded 65% Remazol Brilliant Blue–R (RBBR) dye within 72 h using the only laccase. This study shows that laccase may have a major role in synthetic dyes' decolorization process, followed by MnP and LiP.

Species Listing of Macrofungi Found in Paracelis Mountain Province, Philippines

Mushrooms are an important natural source of food and medicine. In the Philippines, only a few studies have been conducted on the diversity of mushrooms especially in the mountainous areas. The present study was conducted to document the species of macrofungi found in Paracelis, Mountain Province. The knowledge gained from this study can reveal their importance to the community. A total of 37 macrofungi belonging to 16 families, 26 genera, and 29 species were collected and identified. Twenty nine of the collected macrofungi were identified up to its species level and eight were only identified at its genus level. The collected samples were subjected to morphological identification based on its macroscopic and microscopic characteristics. The identified macrofungi were: Auricularia auricula-judae, Conocybe arrhenii, Coprinellus disseminatus, Coprinus sp., Crepidotus mollis, Daldinia concentrica, Earliella scabrosa, Favolus acervatus, Fomes fomentarium, Ganoderma applanatum, Ganoderma fornicatum, Ganoderma lucidum, Hygrocybe sp., Irpex lacteus, Lentinus strigosus, Lenzites elegans, Lepiota lilacea, Lepiota sp., Marasmiellus ramealis, Microporus xanthopus, Mycena sp., Panellus mitis, Paneolus cyanescens, Parasola plicalitis, Psathyrella candolleana, Psathyrella sp., Russula sp. Schizophyllum commune, Trametes elegans, Trametes gibbosa, Trametes hirsuta, Trametes versicolor, Trametes sp. 1, Trametes sp. 2, and Xylaria papulis. Out of these macrofungi, four species were identified as edible, viz: Auricularia auricula, Lentinus strigosus, Coprinus disseminatu, and Schizophyllum commune.

Antifungal activity and mechanism of the essential oils from Litsea (Litsea cubeba), Melissa (Melissa officinalis), Palmarosa (Cymbopogon martini) and Verbena (Verbena officinalis) and their major active constituents against Trametes hirsuta and Laetiporus sulphureus

Antifungal activities of 37 essential oils (EOs) against two wood-decaying fungi, Trametes hirsuta and Laetiporus sulphureus were screened in vitro, and investigated the underlying mechanism. Of the 37 EOs, litsea (Litsea cubeba), melissa (Melissa officinalis), palmarosa (Cymbopogon martini), and verbena (Verbena officinalis) demonstrated strong antifungal activity, in which litsea oil exhibited the strongest antifungal property against T. hirsuta and L. sulphureus, with IC50 values of 72.3 and 40.2 µg/ml, respectively. The compositions of litsea, melissa, palmarosa, and verbena EOs were analyzed using a gas chromatography-mass spectrometry method and demonstrated geranial, geraniol, neral, and citral as their major active constituents. Among of them geranial exhibited the strongest antifungal activity against T. hirsuta and L. sulphureus, with IC50 values of 56.6 and 33.3 µg/ml, respectively. These EOs and their major active constituents increased the plasma membrane permeability of T. hirsuta and L. sulphureus, resulting in the leakage of nucleic acid, protein, and soluble sugar. Results indicate that the EOs of litsea, melissa, palmarosa, and verbena and its major constituents inhibited T. hirsuta and L. sulphureus growth by targeting its plasma membrane.