Endophytic fungi associated with Brucea mollis Wall. ex Kurz.: a hidden source of antimicrobial and antioxidant metabolites.

Geosmithia pallida (KU693285) was isolated from Brucea mollis an endangered medicinal plant of North-East India. The secondary metabolites, produced by the endophytic fungi, extracted by ethyl acetate were screened for antimicrobial activity. G. pallida extract displayed the highest antimicrobial activity against Candida albicans with a minimum inhibitory concentration of 80.5 ± 1.25 µg/mL. G. pallida also showed the highest antioxidant activity which differed insignificantly from Penicillium sp. (P > 0.05). The G. pallida extract also exhibited the highest cellulase activity and also amylase and protease activities. The cytotoxicity assay of the ethyl acetate extract of this endophyte showed negligible effect (1.93 ± 0.42%) on chromosomal aberration as compared to the control (cyclophosphamide monohydrate) (7.20 ± 1.51%). The internal transcribed spacer rDNA sequence of G. pallida was submitted to the NCBI (Accession number KU693285) from India for the first time. The FT-IR spectrophotometry of the bioactive metabolite of G. pallida showed the presence of different functional groups such as alcohol, carboxylic acids, amines, aromatics, alkyl halides, aliphatic amines and alkynes. The GC-MS analysis revealed the presence of acetic acid, 2-phenylethyl ester; tetracosane; cyclooctasiloxane hexadecamethyl; cyclononasiloxane octadecamethyl; octadecanoic acid; phthalic acid, di(2-propylpentyl) ester and nonadecane, 2,6,10,14,18-pentamethyl as the major compounds in the metabolite. The findings of the present work indicated G. pallida as a potential source of important biomolecules without mammalian cytotoxic effects, which can be utilized for pharmaceutical purposes.

Mechanistic Understanding of Gordonia sp. in Biodesulfurization of Organosulfur Compounds.

Although conventional oil refining process like hydrodesulfurization (HDS) is capable of removing sulfur compounds present in crude oil, it cannot desulfurize recalcitrant organosulfur compounds such as dibenzothiophenes (DBTs), benzothiophenes (BTs), etc. Biodesulfurization (BDS) is a process of selective removal of sulfur moieties from DBT or BT by desulfurizing microbes. Therefore, BDS can be used as a complementary and economically feasible technology to achieve deep desulfurization of crude oil without affecting the calorific value. In the recent past, members of biodesulfurizing actinomycete genus Gordonia, isolated from versatile environments like soil, activated sludge, human beings etc. have been greatly exploited in the field of petroleum refining technology. The bacterium Gordonia sp. is slightly acid-fast and has been used for unconventional but potential oil refining processes like BDS in petroleum refineries. Gordonia sp. is unique in a way, that it can desulfurize both aliphatic and aromatic organosulfurs without affecting the calorific value of hydrocarbon molecules. Till date, approximately six different species and nineteen strains of the genus Gordonia have been recognized for BDS activity. Various factors such as enzyme specificity, availability of essential cofactors, feedback inhibition, toxicity of organic pollutants and the oil-water separations limit the desulfurization rate of microbial biocatalyst and influence its commercial applications. The current review selectively highlights the role of this versatile genus in removing sulfur from fossil fuels, mechanisms and future prospects on sustainable environment friendly technologies for crude oil refining.

Bacterial communities and their bioremediation capabilities in oil-contaminated agricultural soils.

Rapid industrialization and development in petrochemical industries have resulted in increased hydrocarbon pollution causing substantial damage to the natural ecosystems including agricultural soils. In the recent, past efforts have been made to treat the contaminated soils using microorganisms by natural processes. Soil bacteria, known for their potential to degrade the soil contaminants, play a vital role in maintaining soil health. In the current study, we observed the influence of hydrocarbon contamination on the physicochemical characteristics and enzymatic activities of the soil. Proteobacteria (30.48%), Actinobacteria (13.91%), and Acidobacteria (12.57%) flourished in the non-contaminated soil whereas contaminated sites were dominated by Proteobacteria (44.02 ± 15.65%). In contrast, the sites experiencing the different degrees of exposure to the hydrocarbon pollution allowed specific augmentation of bacterial taxa (in decreasing order of exposure time), viz. Proteobacteria (60.47%), Firmicutes (32.48%), and Bacteroidetes(13.59%), based on culture-independent approach that suggested their potential role in hydrocarbon degradation as compared to the non-contaminated site. The imputation of metabolic function also supported the positive correlation to the exposure to hydrocarbon pollution, with site 2 being highly abundant for gene families involved in xenobiotics biodegradation. The study provides insights into bacterial community structure with special emphasis on their efficiency to degrade hydrocarbons. The results from the study can help in designing appropriate biodegradation strategies to mitigate the serious problems of oil contamination in agricultural soil.

Isolation and Characterization of an Endophytic Fungus Colletotrichum coccodes Producing Tyrosol From Houttuynia cordata Thunb. Using ITS2 RNA Secondary Structure and Molecular Docking Study.

An endophytic fungus isolated from healthy leaf tissues of Houttuynia cordata Thunb., an ethnomedicinal plant of North East India, showed a considerable amount of antimicrobial activity. The ethyl acetate extract of the fungal culture filtrates displayed promising antimicrobial activity against a panel of clinically significant pathogens including Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Bioassay guided purification of the organic extract using column and thin layer chromatography yielded a pure homogenous compound which was identified using spectroscopic methods (essentially by 1H NMR and MS) as tyrosol, a well-known phenylethanoid present in several natural sources. Besides, molecular docking studies against tyrosyl tRNA synthetases (TyrRS) of S. aureus (PDB ID: 1JIL) and E. coli (PDB ID: 1VBM), and CYP45014α-lanosterol demethylase (CYP51) of C. albicans (PDB ID: 5FSA) revealed tyrosol has a strong binding affinity with the enzyme active site residues. The fungus was identified as Colletotrichum sp. and characterized by its genomic ITS rDNA and ITS2 sequences. Phylogenetic analyses showed clustering of our isolate with Colletotrichum coccodes. Species of Colletotrichum are also reported to be plant pathogens. Therefore, to confirm the endophytic lifestyle of the isolate, ITS2 RNA secondary structure study was undertaken. The result indicated our isolate exhibited differences in the folding pattern as well as in motif structures when compared to those of pathogenic C. coccodes. The findings indicated that endophytic fungi harboring H. cordata could be explored as a potent source of antimicrobial agents.

Efficacy of Mentha spicata essential oil in suppression of Aspergillus flavus and aflatoxin contamination in chickpea with particular emphasis to mode of antifungal action.

The present study reports in vivo antifungal and antiaflatoxigenic efficacy of Mentha spicata essential oil (EO) against toxigenic Aspergillus flavus strain LHP(C)-D6 in chickpea food system up to 12 months of storage. In addition, the mode of antifungal action of EO was also determined to understand the mechanism of fungal growth inhibition. The in vivo study with different concentrations of M. spicata EO showed dose-dependent decrease in fungal colony count as well as aflatoxin B1 concentration. The EO caused >50% protection in inoculated sets and >70% protection in uninoculated sets of chickpea food system against A. flavus at 1.0 µL mL(-1) air concentration. However, at the same concentration, EO caused 100% inhibition to aflatoxin B1 production in both sets when analyzed through high-performance liquid chromatography (HPLC). The antifungal target of EO in fumigated cells of A. flavus was found to be the plasma membrane when analyzed through electron microscopic observations and ions leakage test. The EO fumigated chickpea seeds showed 100% seed germination and seedling growth after 12 months of storage. Based on these observations, M. spicata EO can be recommended as plant-based preservative for safe protection of food commodities during storage conditions against fungal and most importantly mycotoxin contaminations.