Microbial Oil Research Articles

Land-use effects on leaf-litter breakdown in streams in a tropical lowland catchment

The expansion of oil palm and cattle grazing in the tropics continues to alter numerous ecosystem functions. The generated land-use change is potentially impacting stream leaf-litter breakdown, a fundamental process for freshwater ecosystems. To assess the effect of land-use change, we studied breakdown rates of forest (Pachira aquatica, Pouroma aspera, Sloanea ampla, and Hippocratea volubilis) and oil palm (Elaeis guineensis) leaves after a 26-day immersion in streams surrounded by rainforests, grazing lands, or oil palm plantations with and without riparian buffers. In addition, we assessed breakdown drivers by deploying litter bags (248) of two mesh sizes (15 mm and 0.5 mm) allowing or restricting macroinvertebrates’ access (134 coarse-mesh bags and 114 fine-mesh bags). Forest leaf breakdown by microbes (fine-mesh) was lower by 55% in the plantations compared to forests; while microbial oil palm-leaf breakdown was lower by 32% in the unbuffered plantations compared to forests (p < 0.05). Total litter breakdown was lower (p > 0.05) in the plantations but not when these preserved riparian buffers. Litter breakdown was driven primarily by microbes in all land uses except in the buffered plantations, possibly due to increased shredder biomass. These results suggest that oil palm agriculture may decrease microbial stream leaf-litter breakdown, especially in streams with no riparian buffers.

Non-sterile cultivation of oleaginous organisms

Cultivating oleaginous organisms in non-sterile conditions can reduce the energy and cost of microbial oil production. Recent studies use strategies that enable non-sterile cultivation without affecting bioprocess productivity. This forum discusses the trends, strategies, and prospects of non-sterile cultivation, as successful non-sterile cultivation could make microbial oil production economically viable.

Enhancing the biosynthesis of polyunsaturated fatty acids by Rhodotorula mucilaginosa and Lodderomyces elongisporus

Single-cell oils (SCO) produced by oleaginous yeast hold promise as a sustainable alternative for producing nutritionally and pharmaceutically valuable lipids. However, the accumulation of oils varies substantially between yeast spp. Consequently, identifying well-suited producers with a high innate capacity for lipids biosynthesis is paramount. Equally important is optimizing culturing and processing conditions to realize the total lipids production potential of selected strains. The marine Rhodotorula mucilaginosa and Lodderomyces elongisporus yeast were investigated to explore their potential for polyunsaturated fatty acids (PUFAs) production on high glucose media (HGM) using two-stage culture mode. Both strains accumulated > 20% (w/w) of their dry cell weight as lipids when grown on HGM using a two-stage culture system. Both yeast isolates exhibited a maximal lipid/biomass coefficient (YL/X) of 0.58–0.66 mg/mg at 7 °C and 0.49–0.53 mg/mg at 26 °C when grown on 8% glucose and produced monounsaturated and PUFAs similar to that of Menhaden and Salmon marine oils. For the first time, significant amounts of Eicosapentaenoic acid (19%) and Eicosadienoic acid (19.6%) were produced by L. elongisporus and R. mucilaginosa, respectively. Thus, the SCO derived from these wild strains possesses significant potential as a substitute source for the industrial-scale production of long-chain PUFAs, making them a promising contender in the market.Graphical

The Multifaceted Role of Microbial Biosurfactants in Various Fields

This review paper aims to explore the multifaceted roles and applications of microbial surfactants, also known as biosurfactants. These amphiphilic compounds, synthesized by various microorganisms such as Acinetobacter sp., Bacillus sp., Candida antarctica, and Pseudomonas aeruginosa, exhibit both hydrophobic and hydrophilic properties. Through lowering surface tension and interfacial tension, biosurfactants facilitate fluid phase aggregation, imparting organisms with unique physiological and ecological advantages. The synthesis of biosurfactants in bacteria not only contributes to antimicrobial activity but also enhances substrate availability for cellular absorption, particularly in challenging environmental conditions. Categorized based on molecular weight and chemical composition, biosurfactants offer numerous benefits including biodegradability, reduced toxicity, and versatile applications in agriculture, detergent production, biopesticides, and microbial oil enhancement processes. This review synthesizes current knowledge on microbial surfactants, elucidating their diverse roles and promising applications across various industries.

Sugarcane bagasse valorization through integrated process for single cell oil, sulfonated carbon-based catalyst and biodiesel co-production

This study demonstrates the conversion of sugarcane bagasse (SB) into single cell oil (SCO), sulfonated carbon-based catalyst and biodiesel; this process aligns with waste-to-energy and circular bioeconomy concepts. SB was treated with dilute sulfuric acid to achieve SB hydrolysate (SBH) and SB solid residue (SBS). Candida tropicalis KKU-NP1, a newly isolated yeast, accumulated SCO content of 26.5 % from undetoxified SBH medium. A novel sulfonated carbon-based catalyst (SBS@SC) was generated from SBS by a one-step hydrothermal sulfonation process. It showed significant catalytic activity for the conversion of SCO-rich KKU-NP1 wet cell into biodiesel (FAME) under direct transesterification optimal conditions, with a FAME conversion yield of 90.1 %. Based on FAME profile, most of the estimated physicochemical and fuel properties of FAME were within the limits of ASTMD6751 and EN 14214 for biodiesel standards. For integrated process the final production of about 12.0 g SCO, 606.3 g SBS@SC catalyst and 10.8 g biodiesel from 1000 g raw SB were achieved.This study highlights the utilization of SB as a low-cost feedstock for producing multiple value-added products, emphasizing the advantages of waste utilization by integrated biorefinery concept, yielding practically no waste by-products over the whole production process.

Valorization of Sugarcane Vinasse and Crude Glycerol for Single-Cell Oils Production by Rhodotorula glutinis R4: A Preliminary Approach to the Integration of Biofuels Industries for Sustainable Biodiesel Feedstock

Single-cell oils (SCOs) offer a promising alternative to conventional biodiesel feedstocks. The main objective of this work was to obtain SCOs suitable for biodiesel production from the oleaginous yeast Rhodotorula glutinis R4 using sugarcane vinasse from a local sugar-derived alcohol industry as the substrate. Additionally, crude glycerol from the local biodiesel industry was evaluated as a low-cost carbon source to replace expensive glucose and as a strategy for integrating the bioethanol and biodiesel industries for the valorization of both agro-industrial wastes. R4 achieved a high lipid accumulation of 88% and 60% (w/w) in vinasse-based culture media, containing 10% and 25% vinasse with glucose (40 g L−1), respectively. When glucose was replaced with crude glycerol, R4 showed remarkable lipid accumulation (40%) and growth (12.58 g L−1). The fatty acids profile of SCOs showed a prevalence of oleic acid (C18:1), making them suitable for biodiesel synthesis. Biodiesel derived from R4 oils exhibits favorable characteristics, including a high cetane number (CN = 55) and high oxidative stability (OS = 13 h), meeting international biodiesel standards (ASTMD6751 and EN14214) and ensuring its compatibility with diesel engines. R. glutinis R4 produces SCOs from vinasse and crude glycerol, contributing to the circular economy for sustainable biodiesel production.

Exploring the potential of microbial biomass and microbial extracted oils in tribology: a sustainable frontier for environmentally acceptable lubricants

ABSTRACT Mineral oil-derived lubricants, extracted from fossil fuels, account for approximately 90% of the lubricant market. A large proportion of these lubricants end up in the environment through usage, spillage, and disposal, leading to contamination of aquatic systems, ecosystems, and agricultural lands. To address this, new regulations were released (e.g. Vessel General Permit 2013) to promote the use of Environmentally Acceptable Lubricants (EALs) over conventional, toxic, non-biodegradable mineral oils. Today, the range of EAL is limited, particularly affecting the maritime sector. Since 2013, the variety and effectiveness of EALs have improved, but further advancements and cost reductions are essential. This study focuses on developing sustainable bio-based additives from microbial processes to enhance EALs. These additives, sourced through fermentation, avoid using fossil fuels and do not require arable land, preserving water resources and food production areas. The research reveals that yeast, high in sulfur and phosphorus, and microbial oils, mainly carboxylic acids, effectively stabilize EAL formulations, reducing friction and wear in water-based lubricants. Microbial oils are superior in reducing friction, while yeast offers better wear protection. This study opens the possibility of incorporating various bio-based products into EALs, providing a sustainable, environmentally friendly option.

Scale-up of microbial lipid and bioethanol production from oilcane

Microbial oils are a sustainable biomass-derived substitute for liquid fuels and vegetable oils. Oilcane, an engineered sugarcane with superior feedstock characteristics for biodiesel production, is a promising candidate for bioconversion. This study describes the processing of oilcane stems into juice and hydrothermally pretreated lignocellulosic hydrolysate and their valorization to ethanol and microbial oil using Saccharomyces cerevisiae and engineered Rhodosporidium toruloides strains, respectively. A bioethanol titer of 106 g/L was obtained from S. cerevisiae grown on oilcane juice in a 3 L fermenter, and a lipid titer of 8.8 g/L was obtained from R. toruloides grown on oilcane hydrolysate in a 75 L fermenter. Oil was extracted from the R. toruloides cells using supercritical CO2, and the observed fatty acid profile was consistent with previous studies on this strain. These results demonstrate the feasibility of pilot-scale lipid production from oilcane hydrolysate as part of an integrated bioconversion strategy.

Monitoring Yeast Cultures Grown on Corn Stover Hydrolysate for Lipid Production

Microbial oils can be used as an alternative sustainable and renewable feedstock to fossil reserves for producing lubricants and polyurethane materials. Two oleaginous yeasts were grown on non-detoxified corn stover hydrolysate supplemented with corn steep liquor and mineral medium in shake flasks. Trichosporon oleaginosus DSM 11815 displayed the highest lipid production. This strain was further cultivated in a bench bioreactor, using the same culture medium, under a batch regime. Flow cytometry was used to monitor the T. oleaginosus culture using the dual staining technique (SYBR Green and PI) for cell membrane integrity detection. Values of 42.28% (w/w) and 0.06 g/Lh lipid content and lipid productivity, respectively, were recorded for T. oleaginosus cultivated in the bench bioreactor operated under a batch regime. During the cultivation, most of the yeast cells maintained their integrity. T. oleaginosus has the potential to be used as an oil microbial source for a wide range of industrial applications. In addition, it is robust in adverse conditions such as lignocellulosic hydrolysate exposure and oxygen-limiting conditions. Flow cytometry is a powerful and useful tool for monitoring yeast cultivations on lignocellulosic hydrolysates for cell count, size, granularity, and membrane integrity detection.

Identification of lipid synthesis genes in Schizochytrium sp. and their application in improving eicosapentaenoic acid synthesis in Yarrowia lipolytica

BackgroundEicosapentaenoic acid (EPA) is widely used in the functional food and nutraceutical industries due to its important benefits to human health. Oleaginous microorganisms are considered a promising alternative resource for the production of EPA lipids. However, the storage of EPA in triglyceride (TG) becomes a key factor limiting its level.ResultsThis study aimed to incorporate more EPA into TG storage through metabolic engineering. Firstly, key enzymes for TG synthesis, the diacylglycerol acyltransferase (DGAT) and glycerol-3-phosphate acyltransferase (GPAT) genes from Schizochytrium sp. HX-308 were expressed in Yarrowia lipolytica to enhance lipid and EPA accumulation. In addition, engineering the enzyme activity of DGATs through protein engineering was found to be effective in enhancing lipid synthesis by replacing the conserved motifs “HFS” in ScDGAT2A and “FFG” in ScDGAT2B with the motif “YFP”. Notably, combined with lipidomic analysis, the expression of ScDGAT2C and GPAT2 enhanced the storage of EPA in TG. Finally, the accumulation of lipid and EPA was further promoted by identifying and continuing to introduce the ScACC, ScACS, ScPDC, and ScG6PD genes from Schizochytrium sp., and the lipid and EPA titer of the final engineered strain reached 2.25 ± 0.03 g/L and 266.44 ± 5.74 mg/L, respectively, which increased by 174.39% (0.82 ± 0.02 g/L) and 282.27% (69.70 ± 0.80 mg/L) compared to the initial strain, respectively.ConclusionThis study shows that the expression of lipid synthesis genes from Schizochytrium sp. in Y. lipolytica effectively improves the synthesis of lipids and EPA, which provided a promising target for EPA-enriched microbial oil production.

Evaluating oleaginous yeasts for enhanced microbial lipid production using sweetwater as a sustainable feedstock

BackgroundYeasts exhibit promising potential for the microbial conversion of crude glycerol, owing to their versatility in delivering a wide range of value-added products, particularly lipids. Sweetwater, a methanol-free by-product of the fat splitting process, has emerged as a promising alternative feedstock for the microbial utilization of crude glycerol. To further optimize sweetwater utilization, we compared the growth and lipid production capabilities of 21 oleaginous yeast strains under different conditions with various glycerol concentrations, sweetwater types and pH.ResultsWe found that nutrient limitation and the unique carbon composition of sweetwater boosted significant lipid accumulation in several strains, in particular Rhodosporidium toruloides NRRL Y-6987. Subsequently, to decipher the underlying mechanism, the transcriptomic changes of R. toruloides NRRL Y-6987 were further analyzed, indicating potential sugars and oligopeptides in sweetwater supporting growth and lipid accumulation as well as exogenous fatty acid uptake leading to the enhanced lipid accumulation.ConclusionOur comparative study successfully demonstrated sweetwater as a cost-effective feedstock while identifying R. toluroides NRRL Y-6987 as a highly promising microbial oil producer. Furthermore, we also suggested potential sweetwater type and strain engineering targets that could potentially enhance microbial lipid production.Graphical abstract

Combination of alkaline biodiesel-derived crude glycerol pretreated corn stover with dilute acid pretreated water hyacinth for highly-efficient single cell oil production by oleaginous yeast Cutaneotrichosporon oleaginosum

Single cell oil (SCO) prepared from biodiesel-derived crude glycerol (BCG) and lignocellulosic biomass (LCB) via oleaginous yeasts is an intriguing alternative precursor of biodiesel. Here, a novel strategy combining alkaline BCG pretreated corn stover and dilute acid pretreated water hyacinth for SCO overproduction was developed. The mixed pretreatment liquors (MPLs) were naturally neutralized and adjusted to a proper carbon-to-nitrogen ratio beneficial for SCO overproduction by Cutaneotrichosporon oleaginosum. The toxicity of inhibitors was relieved by dilution detoxification. The enzymatic hydrolysate of solid fractions was suitable for SCO production either separately or simultaneously with MPLs. Fed-batch fermentation of the MPLs resulted in high cell mass, SCO content, and SCO titer of 80.7 g/L, 75.7 %, and 61.1 g/L, respectively. The fatty acid profiles of SCOs implied high-quality biodiesel characteristics. This study offers a novel BCG&LCB-to-SCO route integrating BCG-based pretreatment and BCG/LCB hydrolysates co-utilization, which provides a cost-effective technical route for micro-biodiesel production.

Valorization of durian peel as a carbon feedstock for a sustainable production of heterogeneous base catalyst, single cell oil and yeast-based biodiesel

The present study reports a successful attempt to produce single cell oil (SCO), heterogeneous base catalyst and yeast-based biodiesel from durian peel as a promising carbon feedstock by means of the waste-to-energy concept. For this purpose, first, durian peel (DP) was hydrolyzed by dilute sulfuric acid to obtain xylose-rich DP hydrolysate (XDPH) and post-hydrolysis DP solid residue (DPS). Candida viswanathii PSY8, a newly isolated oleaginous yeast, showed high SCO accumulation (5.1 ± 0.1 g/L) and SCO content (35.3 ± 0.13 %) on undetoxified XDPH medium. A novel heterogeneous base catalyst (DPS-K) prepared from DPS by wet impregnation technique with KOH, exhibited considerable catalytic activity to convert SCO-rich wet yeast of C. viswanathii PSY8 into yeast-based biodiesel (FAME) via direct transesterification with a maximum FAME yield of 94.3 % under optimal conditions (6 wt% catalyst, 10:1 methanol to wet yeast ratio, 75 °C, and 2 h). Moreover, most of the yeast-based biodiesel properties obtained from the FAME profiles were correlated well with the biodiesel standards limit of Thai, ASTMD6751 and EN 14214. Additionally, the energy output of FAME produced about 37.5 MJ/kg was estimated. Thus, this present finding demonstrated the favorable strategy for sustainable and eco-friendly production of new generation biodiesel.

Rhodotorula kratochvilovae outperforms Cutaneotrichosporon oleaginosum in the valorisation of lignocellulosic wastewater to microbial oil

Rhodotorula kratochvilovae has shown to be a promising species for microbial oil production from lignin-derived compounds. Yet, information on R. kratochvilovae’s detoxification and microbial oil production is scarce. This study investigated the growth and microbial oil production on the phenolic-containing effluent from poplar steam explosion and its detoxification with five R. kratochvilovae strains (EXF11626, EXF9590, EXF7516, EXF3697, EXF3471) and compared them with Cutaneotrichosporon oleaginosum. The R. kratochvilovae strains reached a maximum growth rate up to four times higher than C. oleaginosum. Furthermore, all R. kratochvilovae strains generally degraded phenolics more rapidly and to a larger extent than C. oleaginosum. However, the diluted substrate limited the lipid production by all strains as the maximum lipid content and titre were 10.5% CDW and 0.40 g/L, respectively.

Sustainable production of lipids from cocoa fatty acid distillate fermentation driven by adaptive evolution in Yarrowia lipolytica

Single cell oil production using oleaginous yeasts is a promising alternative to animal and plant-derived lipids. But substrate costs for microbial fermentation are a major bottleneck. Using side streams as alternative to substrates like glucose, for growing yeast, is a potential cost-effective solution. By combining a previously reported process of growing yeasts on a solid cocoa fatty acid distillate side stream with adaptive evolution techniques, the growth of oleaginous yeast Yarrowia lipolytica was improved by 2-fold. The lipid titre was also boosted by more than 3-fold. Using transcriptomics, key genes were identified that are possibly involved in tailoring of lipid composition, side stream utilisation and enhancement of lipid titres. Candidate genes were also identified that might enable efficient growth and utilization of fatty acids and triacylglycerides found in cocoa fatty acid distillate. In summary, this research has improved the understanding of side stream utilisation for lipid production in oleaginous yeast.

Fruit residues as substrates for single-cell oil production by Rhodococcus species: physiology and genomics of carbohydrate catabolism.

Strains belonging to R. opacus, R. jostii, R. fascians, R. erythropolis and R. equi exhibited differential ability to grow and produce lipids from fruit residues (grape marc and apple pomace), as well as single carbohydrates, such as glucose, gluconate, fructose and sucrose. The oleaginous species, R. opacus (strains PD630 and MR22) and R. jostii RHA1, produced higher yields of biomass (5.1-5.6gL-1) and lipids (38-44% of CDW) from apple juice wastes, in comparison to R. erythropolis DSM43060, R. fascians F7 and R. equi ATCC6939 (4.1-4.3gL-1 and less than 10% CDW of lipids). The production of cellular biomass and lipids were also higher in R. opacus and R. jostii (6.8-7.2gL-1 and 33.9-36.5% of CDW of lipids) compared to R. erythropolis, R. fascians, and R. equi (3.0-3.6gL-1 and less than 10% CDW of lipids), during cultivation of cells on wine grape waste. A genome-wide bioinformatic analysis of rhodococci indicated that oleaginous species possess a complete set of genes/proteins necessary for the efficient utilization of carbohydrates, whereas genomes from non-oleaginous rhodococcal strains lack relevant genes coding for transporters and/or enzymes for the uptake, catabolism and assimilation of carbohydrates, such as gntP, glcP, edd, eda, among others. Results of this study highlight the potential use of the oleaginous rhodococcal species to convert sugar-rich agro-industrial wastes, such as apple pomace and grape marc, into single-cell oils.

Dietary intake of different ratios of ARA/DHA in early stages and its impact on infant development.

Long-chain polyunsaturated fatty acids (LC-PUFAs), arachidonic acid (ARA, 20:4n-6) and docosahexaenoic acid (DHA, 22:6n-3) are essential in the development of infants. ARA and DHA from breast milk or infant formula are the main sources of access for infants to meet their physiological and metabolic needs. The ratio of ARA to DHA in breast milk varies among regions and different lactation stages. Different ratios of ARA and DHA mainly from algal oil, animal fat, fish oil, and microbial oil, are added to infant formula in different regions and infant age ranges. Supplementing with appropriate ratios of ARA and DHA during infancy promotes brain, neural, visual, and other development aspects. In this review, we first introduced the current intake status of ARA and DHA in different locations, lactation stages, and age ranges in breast milk and infant formula. Finally, we discussed the effect of different ratios of ARA and DHA on infant development. This review provided a comprehensive research basis for the nutritional research of infants who consume different ratios of ARA and DHA.

Shotgun-Based Mass Spectrometry Analysis of Phospholipid and Triacylglycerol Molecular Species and Eicosanoids in Salmon Muscle Tissue on Feeding Microbial Oil.

The continuous growth of aquaculture places a growing demand on alternative sources of fish oil (FO). Certain microorganisms provide a sustainable replacement for FO due to their content of EPA and DHA, which are essential for fish health. Appreciable evidence shows that changes in feeding sources may alter the nutritional components of salmon; however, the influence of diets on lipid species remains unclear. In this study, the identification and semi-quantification of lipid molecular species in salmon muscle during feeding with a microbial oil (MO) were carried out by focusing on triacylglycerol (TAG) and diacyl-phospholipid using shotgun-based mass spectrometry analysis. DHA in the MO diet was efficiently incorporated into phospholipid structures on feeding, followed by accumulation in salmon muscle. The MO diet elevated the level of certain EPA-containing TAGs, such as TAG C52:5 (16:0_16:0_20:5) and TAG C54:6 (16:0_18:1_20:5), indicating that the MO diet may be an excellent source for enhancement of the abundance of ω3 lipids. Further, prostaglandins (PGs) PGE2 and PGF3α were identified and quantified for the first time in salmonid tissue.

Mushrooms for enhanced agriculture sustainability – the MUSA concept

The project MUSA – MUshrooms for Sustainable Agriculture is an effort to use mushroom-based processes to enhance agriculture sustainability in Nordic and Baltic countries. The project covers both the production of fruitbodies of edible fungi and the upgrading of the exhausted substrate from mushroom cultivation. The suitability of residues generated locally for producing edible mushrooms is investigated. Residues from Nordic agriculture and sub-utilized streams from forestry management, as well as wood processing by-products, are evaluated as the substrate base for producing shiitake (Lentinula edodes) and oyster (Pleurotus spp.) mushrooms. The project explores the potential of spent mushroom substrate (SMS) to support food production. SMS prospective as source of bioactive compounds and sugars is evaluated. MUSA investigates the suitability of SMS hydrolysates as carbon sources for cultivating oleaginous yeast to produce microbial oil suitable for human consumption. Using SMS for substituting mineral fertilizers and providing wastewater bioremediation solutions is also assessed.

Analysis of technological alternatives for microbial oil production

At present, lignocellulosic biomass and especially agro-industrial by-products have ceased to be considered solid residues, to become potential raw materials for various processes. In this sense, the present work aims to analyze technological alternatives for  obtaining microbial oil in a pilot facility, through simulation in SuperPro Designer simulator, in order to implement a process that is technically, economically and environmentally efficient.  Starting from a previous experimental work, where the best results for the cell growth stage were obtained at a C/N ratio of 100 and a fermentation time of 72 hours. The technological design of a base case of a plant for the production of microbial oil was based on the process designed by Vieira (2016) 1 and simulated as technological alternatives to obtain microbial oil by submerged fermentation (alternative 1) and solid-state fermentation (alternative 2). Technical, economic and environmental analyzes of each technological alternative were performed, obtaining the best results for alternative 2, in which a production of 614.9 liters of oil per year is obtained, a productivity of 2.66 g/Lh, and a unit cost of 224.25 €/L. In addition the filtered residues can be recycled, or can be used as a feedstock for animal feed production.