Poly-Unsaturated Fatty Acids (PUFAs) from Cunninghamella elegans Grown on Glycerol Induce Cell Death and Increase Intracellular Reactive Oxygen Species.

Cunninghamella elegans NRRL-1393 is an oleaginous fungus able to synthesize and accumulate unsaturated fatty acids, amongst which the bioactive gamma-linolenic acid (GLA) has potential anti-cancer activities. C. elegans was cultured in shake-flask nitrogen-limited media with either glycerol or glucose (both at ≈60 g/L) employed as the sole substrate. The assimilation rate of both substrates was similar, as the total biomass production reached 13.0-13.5 g/L, c. 350 h after inoculation (for both instances, c. 27-29 g/L of substrate were consumed). Lipid production was slightly higher on glycerol-based media, compared to the growth on glucose (≈8.4 g/L vs. ≈7.0 g/L). Lipids from C. elegans grown on glycerol, containing c. 9.5% w/w of GLA, were transformed into fatty acid lithium salts (FALS), and their effects were assessed on both human normal and cancerous cell lines. The FALS exhibited cytotoxic effects within a 48 h interval with an IC50 of about 60 µg/mL. Additionally, a suppression of migration was shown, as a significant elevation of oxidative stress levels, and the induction of cell death. Elementary differences between normal and cancer cells were not shown, indicating a generic mode of action; however, oxidative stress level augmentation may increase susceptibility to anticancer drugs, improving chemotherapy effectiveness.

Laboratory- and Pilot-Scale Cultivation of Tetraselmis striata to Produce Valuable Metabolic Compounds.

Marine microalgae are considered an important feedstock of multiple valuable metabolic compounds of high biotechnological potential. In this work, the marine microalga Tetraselmis striata was cultivated in different scaled photobioreactors (PBRs). Initially, experiments were performed using two different growth substrates (a modified F/2 and the commercial fertilizer Nutri-Leaf (30% TN-10% P-10% K)) to identify the most efficient and low-cost growth medium. These experiments took place in 4 L glass aquariums at the laboratory scale and in a 9 L vertical tubular pilot column. Enhanced biomass productivities (up to 83.2 mg L-1 d-1) and improved biomass composition (up to 41.8% d.w. proteins, 18.7% d.w. carbohydrates, 25.7% d.w. lipids and 4.2% d.w. total chlorophylls) were found when the fertilizer was used. Pilot-scale experiments were then performed using Nutri-Leaf as a growth medium in different PBRs: (a) a paddle wheel, open, raceway pond of 40 L, and (b) a disposable polyethylene (plastic) bag of 280 L working volume. Biomass growth and composition were also monitored at the pilot scale, showing that high-quality biomass can be produced, with important lipids (up to 27.6% d.w.), protein (up to 45.3% d.w.), carbohydrate (up to 15.5% d.w.) and pigment contents (up to 4.2% d.w. total chlorophylls), and high percentages of eicosapentaenoic acid (EPA). The research revealed that the strain successfully escalated in larger volumes and the biochemical composition of its biomass presents high commercial interest and could potentially be used as a feed ingredient.

A semi-continuous algal-bacterial wastewater treatment process coupled with bioethanol production.

Harnessing the biomass energy potential through biofuel production offers new outlets for a circular economy. In this study an integrated system which combine brewery wastewater treatment using algal-bacterial aggregates instead of activated sludge was developed. The use of algal-bacterial aggregates can eliminate the aeration requirements and significantly reduce the high biomass harvesting costs associated with algal monocultures. A sequencing batch reactor (SBR) setup operating with and without biomass recirculation was used to investigate pollutant removal rates, aggregation capacity and microbial community characteristics under a range of hydraulic retention times (HRTs) and solid retention times (SRTs). It was observed that biomass recirculation strategy significantly enhanced aggregation and pollutant removal (i.e., 78.7%, 94.2% and 75.2% for d-COD, TKN, and PO43--P, respectively). The microbial community established was highly diverse consisting of 161 Bacterial Operational Taxonomic Units (B-OTUs) and 16 unicellular Eukaryotic OTUs (E-OTUs). Escalation the optimal conditions (i.e., HRT = 4 d, SRT = 10 d) at pilot-scale resulted in nutrient starvation leading to 38-44% w/w carbohydrate accumulation. The harvested biomass was converted to bioethanol after acid hydrolysis followed by fermentation with Saccharomyces cerevisiae achieving a bioethanol production yield of 0.076 g bioethanol/g biomass. These data suggest that bioethanol production coupled with high-performance wastewater treatment using algal-bacterial aggregates is feasible, albeit less productive concerning bioethanol yields than systems exclusively designed for third and fourth-generation biofuel production.

Adaptive laboratory evolution principles and applications in industrial biotechnology.

Adaptive laboratory evolution (ALE) is an innovative approach for the generation of evolved microbial strains with desired characteristics, by implementing the rules of natural selection as presented in the Darwinian Theory, on the laboratory bench. New as it might be, it has already been used by several researchers for the amelioration of a variety of characteristics of widely used microorganisms in biotechnology. ALE is used as a tool for the deeper understanding of the genetic and/or metabolic pathways of evolution. Another important field targeted by ALE is the manufacturing of products of (high) added value, such as ethanol, butanol and lipids. In the current review, we discuss the basic principles and techniques of ALE, and then we focus on studies where it has been applied to bacteria, fungi and microalgae, aiming to improve their performance to biotechnological procedures and/or inspect the genetic background of evolution. We conclude that ALE is a promising and efficacious method that has already led to the acquisition of useful new microbiological strains in biotechnology and could possibly offer even more interesting results in the future.

Single Cell Oil (SCO)-Based Bioactive Compounds: I-Enzymatic Synthesis of Fatty Acid Amides Using SCOs as Acyl Group Donors and Their Biological Activities.

Fatty acid amides (FAAs) are of great interest due to their broad industrial applications. They can be synthesized enzymatically with many advantages over chemical synthesis. In this study, the fatty acid moieties of lipids of Cunninghamella echinulata ATHUM 4411, Umbelopsis isabellina ATHUM 2935, Nannochloropsis gaditana CCAP 849/5, olive oil, and an eicosapentaenoic acid (EPA) concentrate were converted into their fatty acid methyl esters and used in the FAA (i.e., ethylene diamine amides) enzymatic synthesis, using lipases as biocatalysts. The FAA synthesis, monitored using in situ NMR, FT-IR, and thin-layer chromatography, was catalyzed efficiently by the immobilized Candida rugosa lipase. The synthesized FAAs exhibited a significant antimicrobial activity, especially those containing oleic acid in high proportions (i.e., derived from olive oil and U. isabellina oil), against several human pathogenic microorganisms, insecticidal activity against yellow fever mosquito, especially those of C. echinulata containing gamma-linolenic acid, and anticancer properties against SKOV-3 ovarian cancer cell line, especially those containing EPA in their structures (i.e., EPA concentrate and N. gaditana oil). We conclude that FAAs can be efficiently synthesized using microbial oils of different fatty acid composition and used in specific biological applications.

Patterns of Lignocellulosic Sugar Assimilation and Lipid Production by Newly Isolated Yeast Strains From Chilean Valdivian Forest.

Three yeast strains were isolated from decaying wood of Chilean Valdivian forest and identified as Meyerozyma guilliermondii, Scheffersomyces coipomensis, and Sugiyamaella paludigena. These strains were able to efficiently grow on the major monomers contained in Pinus spp. and Eucalyptus spp. wood that includes glucose (Glc), xylose (Xyl), and mannose (Man), showing at 28 °C higher uptake rates for Man, and in some cases for Glc, than for Xyl, used as single carbon sources. Nevertheless, in cultures performed on sugar mixtures, the strains displayed a notable preference for Glc. Additionally, in sugar mixtures, the absence of regulatory mechanisms in sugar assimilation (e.g., catabolic repression) was observed and documented when the activities of several enzymes involved in sugar assimilation (i.e., phosphoglucose isomerase, phosphomannose isomerase, and xylulokinase) were determined. The activity of the key enzymes involved in the onset of lipid accumulation (i.e., NAD+-ICDH) and in fatty acid (FA) biosynthesis (i.e., ATP:CL) indicated a significant accumulation of storage lipids (i.e., up to 24%, w/w) containing oleic and palmitic acids as the major components. The present paper is the first report on the potential of M. guilliermondii, S. coipomensis, and S. paludigena as oleaginous yeasts. We conclude that the new isolates, being able to simultaneously assimilate the major lignocellulosic sugars and efficiently convert them into oily biomass, present a biotechnological potential which deserve further investigation.

High-added value products from microalgae and prospects of aquaculture wastewaters as microalgae growth media.

Aquaculture plays an important role in human nutrition and economic development but is often expanded to the detriment of the natural environment. Several research projects, aimed at cultivating microalgae in aquaculture wastewaters (AWWs) to reduce organic loads and minerals, along with the production of microalgal cell mass and metabolic products, are underway. Microalgal cell mass is of high nutritional value and is regarded as a candidate to replace, partially at least, the fish meal in the fish feed. Also, microalgal cell mass is considered as a feedstock in the bio-fuel manufacture, as well as a source of high-added value metabolic products. The production of these valuable products can be combined with the reuse of AWWs in the light of environmental concerns related with the aquaculture sector. Many research papers published in the last decade demonstrate that plenty of microalgae species are able to efficiently grow in AWWs, mainly derived from fish and shrimp farms, and produce valuable metabolites reducing the AWW pollutant load. We conclude that bio-remediation of AWWs combining with the production of microalgae cell mass and specific metabolites is probably the most convenient and economical solution for AWWs management and can contribute to the sustainable growth of the aquaculture.

Microbial sources of polyunsaturated fatty acids (PUFAs) and the prospect of organic residues and wastes as growth media for PUFA-producing microorganisms.

The discovery of non-fish sources of polyunsaturated fatty acids (PUFAs) is of great biotechnological importance. Although various oleaginous microalgae and fungi are able of accumulating storage lipids (single cell oils - SCOs) containing PUFAs, the industrial applications utilizing these organisms are rather limited due to the high-fermentation cost. However, combining SCO production with other biotechnological applications, including waste and by-product valorization, can overcome this difficulty. In the current review, we present the major sources of fungi (i.e. members of Mucoromycota, fungoid-like Thraustochytrids and genetically modified strains of Yarrowia lipolytica) and microalgae (e.g. Isochrysis, NannochloropsisandTetraselmis) that have come recently to the forefront due to their ability to produce PUFAs. Approaches adopted in order to increase PUFA productivity and the potential of using various residues, such as agro-industrial, food and aquaculture wastes as fermentation substrates for SCO production have been considered and discussed. We concluded that several organic residues can be utilized as feedstock in the SCO production increasing the competitiveness of oleaginous organisms against conventional PUFA producers.

Laboratory evolution strategies for improving lipid accumulation in Yarrowia lipolytica.

Oleaginous microorganisms are of high biotechnological interest being considered as alternative sources of oil (single cell oil-SCO). Current research for increasing productivity of oleaginous microorganisms is focused on the overexpression of genes implicated in lipid synthesis, the inactivation of genes implicated in storage lipid turnover, and on the suppression of competitive to lipid biosynthesis pathways. An alternative strategy, described here, relies on evolution of Yarrowia lipolytica under alternating environments that promote growth, encourage storage lipid synthesis, and reward high energy-containing cells. Derived populations were characterized biochemically, especially on their ability to accumulate lipids, and compared with the starting strain. Interestingly, lipid-accumulating ability early in the evolution was decreased compared with the starting strain. Subsequently, oleaginous lineages dominated, leading to populations able to accumulate lipids in high amounts. A population obtained after 77 generations was able to accumulate 44% w/w of lipid, which was 30% higher than that of the starting strain. We conclude that evolution-based strategies can be utilized as a robust tool for improving lipid accumulation capacity in oleaginous microorganisms.

Sources of microbial oils with emphasis to Mortierella (Umbelopsis) isabellina fungus.

The last years a constantly rising number of publications have appeared in the literature in relation to the production of oils and fats deriving from microbial sources (the "single cell oils"-SCOs). SCOs can be used as precursors for the synthesis of lipid-based biofuels or employed as substitutes of expensive oils rarely found in the plant or animal kingdom. In the present review-article, aspects concerning SCOs (economics, biochemistry, substrates, technology, scale-up), with emphasis on the potential of Mortierella isabellina were presented. Fats and hydrophilic substrates have been used as carbon sources for cultivating Zygomycetes. Among them, wild-type M. isabellina strains have been reported as excellent SCO-producers, with conversion yields on sugar consumed and lipid in DCW values reported comparable to the maximum ones achieved for genetically engineered SCO-producing strains. Lipids produced on glucose contain γ-linolenic acid (GLA), a polyunsaturated fatty acid (PUFA) of high dietary and pharmaceutical importance, though in low concentrations. Nevertheless, due to their abundance in oleic acid, these lipids are perfect precursors for the synthesis of 2nd generation biodiesel, while GLA can be recovered and directed to other usages. Genetic engineering focusing on over-expression of Δ6 and Δ12 desaturases and of C16 elongase may improve the fatty acid composition (viz. increasing the concentration of GLA or other nutritionally important PUFAs) of these lipids.

Data on cellular lipids of Yarrowia lipolytica grown on fatty substrates.

Yarrowia lipolytica, which is model oleaginous yeast with high industrial interest, was cultivated on fatty substrates. Data concerning fatty acid composition of both substrate and yeast lipids and comparisons of the experimental data with model predictions presented in "Biomodification of fats and oils and scenarios of adding value on renewable fatty materials through microbial fermentations: Modelling and trials with Yarrowia lipolytica" (Vasiliadou et al., 2018) were provided. Furthermore, the total yeast lipids were fractionated into their main fractions, that is, phospholipids, glucolipids plus sphingolipids and neutral lipids, and the fatty acid composition of each lipid fraction was reported.

A Leptolyngbya-based microbial consortium for agro-industrial wastewaters treatment and biodiesel production.

A mixed cyanobacterial-mixotrophic algal population, dominated by the filamentous cyanobacterium Leptolyngbya sp. and the microalga Ochromonas (which contributed to the total photosynthetic population with rates of less than 5%), was studied under non-aseptic conditions for its efficiency to remove organic and inorganic compounds from different types of wastes/wastewaters while simultaneously producing lipids. Second cheese whey, poplar sawdust, and grass hydrolysates were used in lab-scale experiments, in photobioreactors that operated under aerobic conditions with different initial nutrient (C, N and P) concentrations. Nutrient removal rates, biomass productivity, and the maximum oil production rates were determined. The highest lipid production was achieved using the biologically treated dairy effluent (up to 14.8% oil in dry biomass corresponding to 124 mg L-1) which also led to high nutrient removal rates (up to 94%). Lipids synthesized by the microbial consortium contained high percentages of saturated and mono-unsaturated fatty acids (up to 75% in total lipids) for all the substrates tested, which implies that the produced biomass may be harnessed as a source of biodiesel.

Critical steps in carbon metabolism affecting lipid accumulation and their regulation in oleaginous microorganisms.

Oleaginous microorganisms are able to convert numerous agro-industrial and municipal wastes into storage lipids (single cell oil (SCO)) and are therefore considered as potential biofuel producers. While from an environmental and technological point of view the idea to convert waste materials into fuels is very attractive, the production cost of SCO is not currently competitive to that of conventional oils due to the low productivity of oleaginous microorganisms in combination with the high fermentation cost. Current strategies used to optimize the lipid-accumulating capacity of oleaginous microorganisms include the overexpression of genes encoding for key enzymes implicated in fatty acid and triacylglycerol synthesis, such as ATP-dependent citrate lyase, acetyl-CoA carboxylase, malic enzyme, proteins of the fatty acid synthase complex, glycerol 3-phosphate dehydrogenase and various acyltransferases, and/or the inactivation of genes encoding for enzymes implicated in storage lipid catabolism, such as lipases and acyl-CoA oxidases. Furthermore, blocking, even partially, pathways competitive to lipid biosynthesis (e.g., those involved in the accumulation of storage polysaccharide or organic acid and polyol excretion) can also increase lipid-accumulating ability in oleaginous microorganisms. Methodologies, such as adaptive laboratory evolution, can be included in existing workflows for the generation of strains with improved lipid accumulation capacity. In our opinion, efforts should be focused in the construction of strains with high carbon uptake rates and a reprogrammed coordination of the individual parts of the oleaginous machinery that maximizes carbon flux towards lipogenesis.

Fish farm effluents are suitable growth media for Nannochloropsis gaditana, a polyunsaturated fatty acid producing microalga.

Fish farm effluents may be used as culture media for marine microalgae, the cell mass of which constitute an excellent fish feed rich in bioactive compounds. In the current investigation different fish farm effluents were tested as culture media for Nannochloropsis strains. Among them, Nannochloropsis gaditana grew well on the effluent released from the sedimentation tank (EST), which is the final step of the wastewater treatment. Mono-algal but non-aseptic cultures were conducted in two types of photo-bioreactors, namely stirred tank reactor (STR) and open pond simulating reactor (OPSR) working under various photoperiods. N. gaditana grew well under full illumination mode on phosphate rich EST in the STR, producing 847.0 mg/L of dry cell mass containing 7.8%, w/w lipids, while when cultivated on phosphate limited EST, cell mass production was slightly lower but lipid biosynthesis was favored, with the lipid content reaching 24.7%, w/w in dry cell mass. In all trials, Nannochloropsis cell mass contained significant quantities of proteins and polysaccharides. Neutral lipids were predominant over polar lipids. Both glycolipid and phospholipid fractions were rich in polyunsaturated fatty acids, especially in eicosapentaenoic acid. We conclude that fish farm wastewaters can be re-used as microalgae growth media, which is of financial and environmental importance.

Storage lipid and polysaccharide metabolism in Yarrowia lipolytica and Umbelopsis isabellina.

Complex biochemical mechanisms are being involved in oleaginous microorganisms during storage lipid and polysaccharide metabolism. Detailed biochemical analyses and monitoring of key enzymes involved in carbon metabolism were performed in Yarrowia lipolytica and Umbelopsis isabellina, which are often used as model oleaginous microorganisms. It was found that during the early oleaginous phase, the carbon source (glucose) was channeled to lipid accumulation, but also to polysaccharide biosynthesis. However, during transition from the early to the late oleaginous phase, glucose was exclusively converted to lipids, while in U. isabellina, but not in Y. lipolytica, an additional conversion of cellular polysaccharides into lipids was observed. After glucose depletion in the growth medium, cellular storage material was degraded either for generating maintenance energy or for supporting further microbial growth, depending on the availability of essential nutrients in the growth medium. We demonstrated that in both microorganisms, reserve lipids were exclusively used as an intra-cellular carbon source in order to generate energy for maintenance purpose. When cellular storage material degradation was related to new cell mass production, a bioconversion of lipids into new lipid-free material, consisting of polysaccharides and proteins, was observed in Y. lipolytica, while new lipid-free material in U. isabellina was richer in proteins. Lipid and polysaccharide remodeling may occur in some cases in both microorganisms. This study revealed some new biochemical features of oleaginous microorganisms that may be crucial for the design of new biotechnological processes, such as the production of bio-molecules of industrial, technological, and medical interest.

Newly isolated yeasts from Tunisian microhabitats: Lipid accumulation and fatty acid composition.

Newly isolated yeasts from different Tunisian microhabitats, such as soil, milk, olive brine, vinegar, and from olive mill wastewater-contaminated biotopes were extensively studied for their biochemical arsenal and morphological features, i.e. cell, ascospore, and lipid body morphology. All strains were classified into the Ascomycota phylum. However, they showed great functional diversity, including different morphological and biochemical features, lipid production ability, and fatty acid profiles. Accordingly, the strains were placed in three different groups: Group I, which includes Candida species; Group II (Pichia and related); and Group III (Kluyveromyces marxianus strain CC1). Group I and II were characterized by a high percentage of oleic acid (41.6-65.3% of total lipids) while in Group III, linoleic acid was the major fatty acid (37.2%). Members of Group I and II were further grouped into subgroups according to their fatty acid composition. Among the newly isolated strains, Pichia etchellsii BM1 was able to accumulate around 25% wt/wt lipid per dry cell mass and thus characterized as oleaginous. Some other strains, such as Candida metapsilosis strain EL2, C. parapsilosis strain LV2, C. pararugosa strain BM24, and K. marxianus strain CC1, which are able to produce extracellular lipases, may be of interest for specific environmental applications and/or for the production of novel lipases.

Rhodosporidium toruloides cultivated in NaCl-enriched glucose-based media: Adaptation dynamics and lipid production.

In the present report and for the first time in the international literature, the impact of the addition of NaCl upon growth and lipid production on the oleaginous yeast Rhodosporidium toruloides was studied. Moreover, equally for first time, lipid production by R. toruloides was performed under nonaseptic conditions. Therefore, the potentiality of R. toruloides DSM 4444 to produce lipid in media containing several initial concentrations of NaCl with glucose employed as carbon source was studied. Preliminary batch-flask trials with increasing amounts of NaCl revealed the tolerance of the strain against NaCl content up to 6.0% w/v. However, 4.0% w/v of NaCl stimulated lipid accumulation for this strain, by enhancing lipid production up to 71.3% w/w per dry cell weight. The same amount of NaCl was employed in pasteurized batch-flask cultures in order to investigate the role of the salt as bacterial inhibiting agent. The combination of NaCl and high glucose concentrations was found to satisfactorily suppress bacterial contamination of R. toruloides cultures under these conditions. Batch-bioreactor trials of the yeast in the same media with high glucose content (up to 150 g/L) resulted in satisfactory substrate assimilation, with almost linear kinetic profile for lipid production, regardless of the initial glucose concentration imposed. Finally, fed-batch bioreactor cultures led to the production of 37.2 g/L of biomass, accompanied by 64.5% w/w of lipid yield. Lipid yield per unit of glucose consumed received the very satisfactory value of 0.21 g/g, a value among the highest ones in the literature. The yeast lipid produced contained mainly oleic acid and to lesser extent palmitic and stearic acids, thus constituting a perfect starting material for "second generation" biodiesel.

Conversion of biodiesel-derived glycerol into biotechnological products of industrial significance by yeast and fungal strains.

Oleochemical activities (e.g. biodiesel production, fat saponification) generate annually very high amounts of concentrated glycerol-containing waters (called crude glycerol) as the principal residues of these processes. Crude glycerol is an industrial residue the valorization of which attracts remarkable and constantly increasing interest. In the current investigation, biodiesel-derived glycerol was employed as substrate for yeast and fungal strains cultivated under nitrogen-limited conditions in shake flasks. Glucose was employed as reference substrate. Several yeasts (Candida diddensiae, Candida tropicalis, Pichia ciferrii, Williopsis saturnus, Candida boidinii, and Candida oleophila) rapidly assimilated glucose and converted it into ethanol, despite aerobic conditions imposed, and were Crabtree-positive. None of these yeasts produced ethanol during growth on glycerol or accumulated significant quantities of lipid during growth on glucose or glycerol. Only Rhodosporidium toruloides produced notable lipid quantities from glucose and to lesser extent from glycerol. Yarrowia lipolytica LFMB 20 produced citrate ≈58 g/L growing on high-glucose media, while on high-glycerol media ≈42 g/L citrate and ≈18 g/L mannitol. During growth on glucose/glycerol blends, glycerol was assimilated first and thereafter glucose was consumed. Fungi produced higher lipid quantities compared with yeasts. High lipid quantities were produced by Mortierella ramanniana, Mucor sp., and mainly Mortierella isabellina, with glycerol being more adequate for M. ramanniana and glucose for Mucor sp. and M. isabellina. M. isabellina ATHUM 2935 produced lipids of 8.5 g/L, 83.3% w/w in dry cell weight (DCW) and conversion yield per unit of glucose consumed ≈0.25 g/g. The respective values on glycerol were 5.4 g/L, 66.6% w/w in DCW and ≈0.22 g/g. Lipids of all microorganisms were analyzed with regards to their fatty acid composition, and M. isabellina presented the closest similitude with rapeseed oil. Crude lipids produced by this fungus and extracted with chloroform/methanol blend, were composed mostly of triacylglycerols, thus indicating that these solvents are adequate for triacylglycerol extraction.

Production of added-value metabolites by Yarrowia lipolytica growing in olive mill wastewater-based media under aseptic and non-aseptic conditions.

Yarrowia lipolytica ACA-YC 5033 was grown on glucose-based media in which high amounts of olive mill wastewaters (OMWs) had been added. Besides shake-flask aseptic cultures, trials were also performed in previously pasteurized media while batch bioreactor experiments were also done. Significant decolorization (∼58%) and remarkable removal of phenolic compounds (∼51% w/w) occurred, with the latter being amongst the highest ones reported in the international literature, as far as yeasts were concerned during their growth on phenol-containing media. In nitrogen-limited flask fermentations the microorganism produced maximum citric acid quantity ≈19.0 g/L [simultaneous yield of citric acid produced per unit of glucose consumed (YCit/Glc)≈0.74 g/g]. Dry cell weight (DCW) values decreased at high phenol-containing media, but, on the other hand, the addition of OMWs induced reserve lipid accumulation. Maximum citric acid concentration achieved (≈52.0 g/L; YCit/Glc≈0.64 g/g) occurred in OMW-based high sugar content media (initial glucose added at ≈80.0 g/L). The bioprocess was successfully simulated by a modified logistic growth equation. A satisfactory fitting on the experimental data occurred while the optimized parameter values were found to be similar to those experimentally measured. Finally, a non-aseptic (previously pasteurized) trial was performed and its comparison with the equivalent aseptic experiment revealed no significant differences. Yarrowia lipolytica hence can be considered as a satisfactory candidate for simultaneous OMWs bioremediation and the production of added-value compounds useful for the food industry.