Bioflocculation of Euglena gracilis via direct application of fungal filaments: a rapid harvesting method

The high cost and environmental impact of traditional microalgal harvesting methods limit commercialization of microalgal biomass. Fungal bioflocculation of microalgae is a promising low-cost, eco-friendly method but the range of fungal and microalgal species tested to date is narrow. Here, eight non-pathogenic, filamentous fungi were screened for their ability to self-pelletize and flocculate Euglena gracilis (ca.50 µm motile microalga) in suspension. Self-pelletization was tested under various rotational speeds, and species which formed pellets (Ø > 0.5 cm) were selected for harvesting tests. Filaments of each species were combined with E. gracilis at various ratios based on dry weight. Harvesting efficiency was determined by measuring the change in cell counts over time, and settling of the flocs was evaluated by batch settling tests. Three fungal species, Ganoderma lucidum, Pleurotus ostreatus, and Penicillium restrictum, were able to reliably flocculate and harvest 62–75% of the microalgae while leaving it unharmed. The results demonstrated that self-pelletization, harvesting, and settling were dependent on the fungal species. The fungi to algae ratio also had significant but contrasting effects on harvesting and settling. In balancing the needs to both harvest and settle the biomass, the optimal fungi to algae ratio was 1:2. The application of fungal filaments to microalgae in suspension produced readily settling flocs and was less time-consuming than other commonly used methods. This method is especially attractive for harvesting microalgal biomass for low-value products where speed, low cost, and cell integrity is vital.

UNUSUALLY LIPID PRODUCTION BY THE MARINE-DERIVED FUNGI Penicillium restrictum

Using marine-derived fungi as a source of lipid production holds promise as an alternative to industrial lipid production for health and nutrition in the future. In the present study, this strain showed a high production of lipid, about plus 80% of the total lipids in the host-derived medium. Gas chromatography analysis of fungal lipids revealed the presence of saturated (mainly palmitic acid C16: 0 and stearic acid C18: 0) and unsaturated fatty acid (mainly linoleic acid C18: 2, oleic acid C18: 1). These findings suggest this marine-derived fungus is a promising source for lipid production in various industrial applications. In particular, a fraction containing glycolipids of the crude extract exhibited potential cytotoxic activity on human oral epidermal carcinoma cell lines. This result is very interesting to further isolate and determine the molecular structure of bioactive glycolipids.

The Effectiveness of Biostimulation, Bioaugmentation and Sorption-Biological Treatment of Soil Contaminated with Petroleum Products in the Russian Subarctic

The effectiveness of different bioremediation methods (biostimulation, bioaugmentation, the sorption-biological method) for the restoration of soil contaminated with petroleum products in the Russian Subarctic has been studied. The object of the study includes soil contaminated for 20 years with petroleum products. By laboratory experiment, we established five types of microfungi that most intensively decompose petroleum hydrocarbons: Penicillium canescens st. 1, Penicillium simplicissimum st. 1, Penicillum commune, Penicillium ochrochloron, and Penicillium restrictum. One day after the start of the experiment, 6 to 18% of the hydrocarbons decomposed: at 3 days, this was 16 to 49%; at 7 days, 40 to 73%; and at 10 days, 71 to 87%. Penicillium commune exhibited the greatest degrading activity throughout the experiment. For soils of light granulometric composition with a low content of organic matter, a more effective method of bioremediation is sorption-biological treatment using peat or granulated activated carbon: the content of hydrocarbons decreased by an average of 65%, which is 2.5 times more effective than without treatment. The sorbent not only binds hydrocarbons and their toxic metabolites but is also a carrier for hydrocarbon-oxidizing microorganisms and prevents nutrient leaching from the soil. High efficiency was noted due to the biostimulation of the native hydrocarbon-oxidizing microfungi and bacteria by mineral fertilizers and liming. An increase in the number of microfungi, bacteria and dehydrogenase activity indicate the presence of a certain microbial potential of the soil and the ability of the hydrocarbons to produce biochemical oxidation. The use of the considered methods of bioremediation will improve the ecological state of the contaminated area and further the gradual restoration of biodiversity.

A Review on Extraction of Lipase from Aspergillus Species and its Applications

Lipase enzyme which catalyses the hydrolysis of fats performs a variety of chemical reactions including esterification, trans-esterification, acidolysis and aminolysis. Lipases are widely used for biotechnological application in dairy industry, oil processing and production of surfactants and preparation of enatiomerically pure pharmaceuticals. Lipases are extracted from plants, animals and microorganisms whereas high amount of lipases are produced by microbes when compared to plants and animal source. Fungi serve as a major source in the production of industrial lipase. Fungi play a vital role in removing hazardous compounds formed by crude oil contamination from oil spill during lipase extraction. Aspergillus flavus, Aspergillus niger, Penicillium restrictum, Rhizomucar rhizopodiformis, Rhizopus oligosporous, Candida rugosa, Rhizopus oryzae, Fusarium oxysporum are few fungal species involved in the production of lipase. Among the species, Aspergillus species produce efficient amount of lipase. Lipase extracted from fungi is more stable and have more diverse properties compared to lipase from other source. Fungal lipases stand out as major source because of their catalytic activity, low cost of production and relative ease in genetic manipulation. This review focuses on the various methods of lipase extraction from Aspergillus species and its applications.

Untargeted Metabolomics Approach for the Discovery of Environment-Related Pyran-2-ones Chemodiversity in a Marine-Sourced Penicillium restrictum

Very little is known about chemical interactions between fungi and their mollusc host within marine environments. Here, we investigated the metabolome of a Penicillium restrictum MMS417 strain isolated from the blue mussel Mytilus edulis collected on the Loire estuary, France. Following the OSMAC approach with the use of 14 culture media, the effect of salinity and of a mussel-derived medium on the metabolic expression were analysed using HPLC-UV/DAD-HRMS/MS. An untargeted metabolomics study was performed using principal component analysis (PCA), orthogonal projection to latent structure discriminant analysis (O-PLSDA) and molecular networking (MN). It highlighted some compounds belonging to sterols, macrolides and pyran-2-ones, which were specifically induced in marine conditions. In particular, a high chemical diversity of pyran-2-ones was found to be related to the presence of mussel extract in the culture medium. Mass spectrometry (MS)- and UV-guided purification resulted in the isolation of five new natural fungal pyran-2-one derivatives—5,6-dihydro-6S-hydroxymethyl-4-methoxy-2H-pyran-2-one (1), (6S, 1’R, 2’S)-LL-P880β (3), 5,6-dihydro-4-methoxy-6S-(1’S, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (4), 4-methoxy-6-(1’R, 2’S-dihydroxy pent-3’(E)-enyl)-2H-pyran-2-one (6) and 4-methoxy-2H-pyran-2-one (7)—together with the known (6S, 1’S, 2’S)-LL-P880β (2), (1’R, 2’S)-LL-P880γ (5), 5,6-dihydro-4-methoxy-2H-pyran-2-one (8), (6S, 1’S, 2’R)-LL-P880β (9), (6S, 1’S)-pestalotin (10), 1’R-dehydropestalotin (11) and 6-pentyl-4-methoxy-2H-pyran-2-one (12) from the mussel-derived culture medium extract. The structures of 1-12 were determined by 1D- and 2D-MMR experiments as well as high-resolution tandem MS, ECD and DP4 calculations. Some of these compounds were evaluated for their cytotoxic, antibacterial, antileishmanial and in-silico PTP1B inhibitory activities. These results illustrate the utility in using host-derived media for the discovery of new natural products.

Drug Leads from Endophytic Fungi: Lessons Learned via Scaled Production

Recently, the isolation and elucidation of a series of polyhydroxyanthraquinones were reported from an organic extract of a solid phase culture of an endophytic fungus, Penicillium restrictum (strain G85). One of these compounds, ω-hydroxyemodin (1), showed promising quorum-sensing inhibition against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) in both in vitro and in vivo models. The initial supply of 1 was 19 mg, and this amount needed to be scaled by a factor of 30 to 50 times, in order to generate material for further in vivo studies. To do so, improvements were implemented to enhance both the fermentation of the fungal culture and the isolation of this compound, with the target of generating > 800 mg of study materials in a period of 13 wk. Valuable insights, both regarding chemistry and mycology, were gained during the targeted production of 1 on the laboratory-scale. In addition, methods were modified to make the process more environmentally friendly by judicious choice of solvents, implementing procedures for solvent recycling, and minimizing the use of halogenated solvents.