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of Cunninghamella echinulata, Mortierella ramanniana, Mucor sp., Thamnidium elegans, and Zygorhynchus moelleri on glycerol and synthesis of lipids
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Several strains of Zygomycetes cultivated on glycerol produced mycelia rich in lipids containing higher amounts of neutral lipids (NL) than glycolipids plus sphingolipids and phospholipids (P), while biosynthesis of P in Mortierella ramanniana, Mucor sp., and Cunninghamella echinulata occurred though NL accumulation process was in progress. Polyuns...
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Context 1
... data of growth and lipid accumulation are presented in Table 1. These data were originated from cultures grown under fully aerobic conditions as in all cases dissolved oxygen concentration in the growth medium was >70% of the saturation value in the first growth steps and declines thereafter reaching to 42% in some of them. ...Context 2
... strain, being able to efficiently convert glycerol into lipid-rich biomass (biomass produced X=8.2 g/l, lipid in biomass L/X>40%-data shown in Fig. 1), could be considered as a potential candidate for glycerol valorization. Other strains, i.e., T. elegans and C. echinulata, also accumulated Culture conditions: growth in flasks, initial substrate concentration 25 g/l, incubation in a rotary shaker at T=28 °C and 180 rpm; initial pH 6. Experiments were performed in duplicate X (g/l) total biomass, L/X (%) lipids in dry biomass, NL neutral lipids, G+S glycolipids plus sphingolipids, P phospholipids noteworthy quantities of storage lipid but the final yields were mediocre due to the restricted growth on glycerol (Table 1). Similarly, high content of reserve lipids have also been reported for Mortierella isabellina grown on high sugar content media [8] and for T. elegans grown on raw glycerol [17]. ...Context 3
... the other hand, the growth on glycerol of Mucor sp. and Z. moelleri was mediocre. In these strains, as well as in C. echinulata, glycerol was taken up with low rates and this was probably the reason for lipid turnover that was observed in these strains (Table 1). In all growth phases fungal mycelia contained higher amounts of NL, that represented the major lipid fraction, than polar lipids (G+S and P; Table 1), a fact which is in accordance with various literature reports on oleaginous microorganisms. ...Context 4
... these strains, as well as in C. echinulata, glycerol was taken up with low rates and this was probably the reason for lipid turnover that was observed in these strains (Table 1). In all growth phases fungal mycelia contained higher amounts of NL, that represented the major lipid fraction, than polar lipids (G+S and P; Table 1), a fact which is in accordance with various literature reports on oleaginous microorganisms. Indeed, NL, in both terms of proportion (% on total lipid) and absolute value (mg/l), increased with time in the cases of C. echinulata, M. ramanniana, and T. elegans, while, in Mucor sp. and Z. moelleri (in which glycerol was not well assimilated) low or nil quantities of NL were synthesized as growth proceeded. ...Citations
... From the obtained results, it can be seen that TDCW production was almost equivalent, irrespective of the trial perform on glucose or glycerol, since, in both instances, X max concentration was achieved c. 330-360 h after inoculation, presenting similar values (≈13.5 g/L) for both trials (Figure 1a). Equally, the substrate assimilation rate was almost the same for the two substrates employed as carbon sources, in disagreement with most literature reports [3,29,[42][43][44][45]] that indicate that for most higher (e.g., Ascomycetes/Basidiomycetes) or lower (e.g., Zygomycetes) fungi, glucose is considered by far to be a more adequate carbon source compared to glycerol, due to the poor regulation of the enzymes implicated in the uptake of glycerol by these very microorganisms [29,43,44]. ...
... From the obtained results, it can be seen that TDCW production was almost equivalent, irrespective of the trial perform on glucose or glycerol, since, in both instances, Xmax concentration was achieved c. 330-360 h after inoculation, presenting similar values (≈13.5 g/L) for both trials (Figure 1a). Equally, the substrate assimilation rate was almost the same for the two substrates employed as carbon sources, in disagreement with most literature reports [3,29,[42][43][44][45]] that indicate that for most higher (e.g., Ascomycetes / Basidiomycetes) or lower (e.g., Zygomycetes) fungi, glucose is considered by far to be a more adequate carbon source compared to glycerol, due to the poor regulation of the enzymes implicated in the uptake of glycerol by these very microorganisms [29,43,44]. Moreover, it was with interest that, in the present investigation, significantly higher lipid production was observed to occur in the trial performed on glycerol (Lmax = 8.4 g/L, simultaneous lipid in TDCW = 62.7% w/w), compared to the experiment performed on glucose (Lmax = 7.0 g/L, simultaneous lipid in TDCW =51.4% w/w), demonstrating, once Moreover, it was with interest that, in the present investigation, significantly higher lipid production was observed to occur in the trial performed on glycerol (L max = 8.4 g/L, simultaneous lipid in TDCW = 62.7% w/w), compared to the experiment performed on glucose (L max = 7.0 g/L, simultaneous lipid in TDCW =51.4% w/w), demonstrating, once more, the suitability of glycerol as the implicated substrate in the conversions performed by C. elegans. ...
... According to the contemporary literature, glycerol in general is not a very appropriate substate amenable to be used for most fungi (higher or lower) due to its poor uptake regulation. For most higher or lower fungi, growth and lipid production on glucose is significantly better than that on glycerol; for instance, the growth of fungi of the genera Aspergillus, Lentinula, Mucor, Morchella, etc., on glycerol was accompanied by lower dry biomass production compared to growth on glucose [27,29,42,43,49]. This is not the case with the growth of oleaginous yeasts (i.e., Rhodosporidium toruloides, Yarrowia lipolytica, Debaryomyces prosopidis, etc.) that in general present excellent growth on glycerol [26,28,50]. ...
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.
... FA alkyl-esters) has resulted in a significant rise in the production of glycerol, which is the main side-product of the trans-esterification reaction process. As a result, an important decrease in the price of glycerol occurred last year, and the topic of its valorization has become crucial for biotechnological industries [2,5,7,8]. The utilization of oleaginous yeasts in the conversion of crude glycerol closes the loop of biodiesel production since the principal residue of this industrial activity (viz. ...
... The utilization of oleaginous yeasts in the conversion of crude glycerol closes the loop of biodiesel production since the principal residue of this industrial activity (viz. crude glycerol) is transformed into triacylglycerols that subsequently will be converted into "second-generation" biodiesel, a process with obvious economic and ecological interest [2,[5][6][7][8]]. ...
Four wild “red” yeast strains (Rhodosporidium kratochvilovae FMCC Y70, R. toruloides NRRL Y-27013, R. toruloides NRRL Y-17902 and R. toruloides NRRL Y-6985) were cultured in shake flasks on industrial glycerol at an initial substrate (Gly0) concentration ≈ 50 g/L under nitrogen limitation. Strains NRRL Y-27013, NRRL Y-17902 and NRRL Y-6985 presented appreciable dry cell weight (DCW) and lipid synthesis (DCW up to 18–19 g/L containing lipids in quantities ≈ 47%, w/w). Strains NRRL Y-27013 and NRRL Y-6985 were further tested in higher Gly0 concentrations (≈90 g/L and ≈110 g/L) with the same initial nitrogen quantity as in the first (“screening”) experiment. Both strains, despite the high Gly0 concentrations and C/N ratios (up to 120 moles/moles) imposed, presented significant DCW production (up to c. 29.0–29.5 g/L). Yeast biomass contained significant lipid (42–43%, w/w) and endopolysaccharide (up to 42%, w/w) quantities. Both lipids and endopolysaccharide quantities (in % w/w) noticeably increased as a response to the imposed nitrogen limitation. Lipids containing mainly oleic and palmitic acids constituted ideal candidates for biodiesel synthesis. Thereafter, the wastewaters derived from the lipid production process (lipid fermentation wastewaters—LFWs) were used as maceration waters in cultivations of edible and medicinal fungi, where novel (non-conventional) substrates were used in the performed cultures. CW (coffee residue + wheat straw), CB (coffee residue + beech wood shavings), OW (olive crop + wheat straw), OB (olive crop + beech wood shavings), RW (rice husk + wheat straw) and RB (rice husk + beech wood shavings) were soaked/sprayed with LFWs or tap water and utilized in the cultivation of Pleurotus, Ganoderma and Lentinula mushrooms. The impact of LFWs on the mycelial growth rate (mm/d) and biomass production was evaluated. The results show that regardless of the wetting method, the highest growth rates (6.2–6.6 mm/d) were noticed on RW and RB for Pleurotus eryngii and Ganoderma resinaceum, on OW, OB and RW for Ganoderma applanatum and on RW, OW and OB for Lentinula edodes. Nevertheless, high biomass production was obtained on substrates soaked with LFWs for Pleurotus ostreatus (RW: 443 mg/g d.w.), L. edodes (RB: 238 mg/g d.w.) and Ganoderma lucidum (RW: 450 mg/g d.w.). Overall, this study demonstrates the possibility of the industrial conversion of low-value agro-waste to mycelial mass and eventually to important food products.
... Fungal lipids have advantages over algal lipids because fungi are fast growing with short span, light independent and can degrade a wide range of carbon sources (Chen et al., 2012). Lipid production by oleaginous fungi using renewable carbon sources such as: glycerol, sewage water, whey and molasses were reported (Bellou et al., 2012;Peng et al., 2013). Oleaginous microorganisms can convert a wide variety of carbon sources into stored lipids (Ratledge, 2004). ...
This study aims to explore microbes from mangroves in Saudi Arabia for their abilities to produce high level of lipids. Mangroves are seldom investigated for oleaginous microbes. A total of 961 isolates of yeasts and filamentous fungi were isolated from 144 submerged marine samples include: 68 decaying leaves of Avicennia marina, 33 decaying thalli of Zostera marina, 14 decaying pneumatophores of Avicennia marina, 9 crab shells, 8 sediment, 7 decaying thalli of Turbinaria ornata and 5 decaying thalli of Cystoseira myrica. Samples were collected from four mangrove sites: Al-Leith, Jeddah and Yanbu at the Red Sea coast and the Syhat mangroves at the Arabian Gulf coast. Isolated fungi were grouped into 62 morphological types that include: 21 yeasts and 41 filamentous fungi. Fifty-four isolates of thraustochytrids were cultured from the four mangrove sites and were grouped into 22 strains. Two oleaginous yeasts: Hortaea werneckii and Rhodotorula mucilaginosa and four Aurantiochytrium strains produced high dry weight ranged between 32 and 49.3 g/L of which 35.2–62 % lipid and their fatty acid profile were determined using GC/MS. Palmitic acid was the major fatty acid in the lipid of the four thraustochytrid isolates and ranged between 5.71 and 82 % of the total fatty acids, 9-Octadecenamide, (Z)- was the major fatty acid amide in the lipid of the two yeast isolates and two thraustochytrid isolates and ranged between 26.94 and 56.63 %, followed by 13-Docosenamide, (Z)- (20.44–34.99 %) from the same four isolates. Other major lipid compounds were: Hexadecanamide (4.35–7.19 %), Cholestrol (7.24–15.07 %), Butylated Hydroxytoluene (3.46–15.76 %), Octadecanamide (3.95–7.9 %), Phenol, 2-(1,1-dimethylethyl)-5-methyl- (1.78–10.33 %) and Pentadecanoic acid (7.47 %).
... For instance, U. isabellina can accumulate unusually high lipid amounts, i.e., up to 80% w/w in dry biomass, under specific growth conditions [10]. Its lipids contain γ-linolenic acid, a polyunsaturated fatty acid (PUFA) of importance for dietary and pharmaceutical purposes [11][12][13] while its more saturated fatty acids can be channeled to the biodiesel production industry [14]. Last but not least, its ability to degrade and/or reduce the toxicity of environmental pollutants (metals, phenolic compounds, etc.) has been reported [15]. ...
... The aforementioned show that U. isabellina, exhibits a clear preference for glucose, a feature that appears to be common among the Mucorales [5,11,[38][39][40]. Furthermore, it is established that Mucorales show poor assimilation ability for glycerol [20,41,42] with the fungus Mortierella ramanniana MUCL 9235 being the only recorded exception [12]. It should be noted, though, that filamentous fungi which belong to the other classification divisions are not very efficient at glycerol uptake as well. ...
... For instance, U. isabellina can accumulate unusually high lipid amounts, i.e., up to 80% w/w in dry biomass, under specific growth conditions [10]. Its lipids contain γ-linolenic acid, a polyunsaturated fatty acid (PUFA) of importance for dietary and pharmaceutical purposes [11][12][13] while its more saturated fatty acids can be channeled to the biodiesel production industry [14]. Last but not least, its ability to degrade and/or reduce the toxicity of environmental pollutants (metals, phenolic compounds, etc.) has been reported [15]. ...
... The aforementioned show that U. isabellina, exhibits a clear preference for glucose, a feature that appears to be common among the Mucorales [5,11,[38][39][40]. Furthermore, it is established that Mucorales show poor assimilation ability for glycerol [20,41,42] with the fungus Mortierella ramanniana MUCL 9235 being the only recorded exception [12]. It should be noted, though, that filamentous fungi which belong to the other classification divisions are not very efficient at glycerol uptake as well. ...
Over the past few years it is observed an increased interest for oleaginous microorganisms in the perspective to produce microbial oils of great commercial interest through the consumption of low/zero cost substrates. In this paper, the physiology of the fungus Umbelopsis isabellina growing on blends of glycerol and glucose was investigated. In all experiments the fungus completely consumed glucose and produced satisfactory quantities of biomass containing reserve lipids in high percentages. However, glycerol concentration in the growth medium was negatively correlated to glucose assimilation rate, mainly during the balanced-growth phase. Nevertheless, at high initial concentrations, glycerol was partially consumed and seemed to contribute positively to the suppression of lipid degradation. Following the discovery of this complex regulatory mechanism regarding glucose and glycerol co-assimilation, the activity of three key-enzymes namely aldolase, glycerol kinase and glycerol dehydrogenase, which are implicated in glycerol and glucose assimilation, was investigated. The experiments revealed a clear preference of the fungus for glucose over glycerol. On the other hand, storage polysaccharides are degraded instead of storage lipid at the late oleaginous phase for maintenance purpose. These new biochemical features will enable the design of appropriate growth media for the co-fermentation of these two substrates by U. isabellina with the aim to maximize lipid accumulation.
... Yarrowia lipolytica yeast was cultivated on raw glycerol whereby its growth was not followed by reserve lipid accumulation but amounts of citric acid were produced [71]. Besides that, oleaginous Zygomycetes converted glycerol into SCO under non-aseptic conditions on selective nitrogen-limited media [72,73]. Yatim et al. [13], Cruz et al. [74], and Rashid et al. [75] have also investigated the production of the mentioned products with glycerol as the sole carbon source. ...
The growing concerns on the rising population, global economy, food safety and environmental pollution have given impetus to the production of polyhydroxyalkanoates (PHAs) and biosurfactant, biodegradable alternatives to petrochemical plastics and surfactants, respectively. However, the application of these compounds is limited due to processing costs. Carbon sources represent half of the PHA and biosurfactant fermentation cost, therefore it is of interest to recycle agro-industrial waste as renewable carbon sources, such as palm oil and sugar cane production waste. This review aimed to provide insight on the utilisation of molasses and sweetwater from sugar cane and palm oil as carbon substrates for PHA and biosurfactant production, as well as providing examples of the application of such compounds. sugar cane and palm oil by-products are feasible as inexpensive feed for the PHA and biosurfactant production. Moreover, since sugar cane and palm oil plants are cultivated in tropical and subtropical areas, utilising these wastes can instil the circular economy spirit in line with the Sustainable Development Goals proposed by the United Nations.
... The addition of emulsifiers to the fermenting medium, on the other hand, raises the overall production cost and has an impact on the final lipid production because it is frequently dissolved in organic solvents, interfering with the estimation of residual hydrophobic substrates. Aurantiochytrium sp By-product of the palm oil industry DHA (190) T. kinney and S. limacinum Crude glycerol (191,192) T. elegans, U. isabellina, C. echinulata, U. ramanniana, Z. moelleri and Mucor sp Crude glycerol GLA (193)(194)(195)(196)(197) Mortierella spp Crude glycerol ARA, DGLA (198) N. gaditana Phosphate-limited wastewaters obtained an aquaculture station EPA (199) Navicula sp., Chlorella sp., Nannochloropsis sp., Dunaliella sp., and Tetraselmis sp. ...
Microbes have gained a lot of attention for their potential in producing polyunsaturated fatty acids (PUFAs). PUFAs are gaining scientific interest due to their important health-promoting effects on higher organisms including humans. The current sources of PUFAs (animal and plant) have associated limitations that have led to increased interest in microbial PUFAs as most reliable alternative source. The focus is on increasing the product value of existing oleaginous microbes or discovering new microbes by implementing new biotechnological strategies in order to compete with other sources. The multidisciplinary approaches, including metabolic engineering, high-throughput screening, tapping new microbial sources, genome-mining as well as co-culturing and elicitation for the production of PUFAs, have been considered and discussed in this review. The usage of agro-industrial wastes as alternative low-cost substrates in fermentation for high-value single-cell oil production has also been discussed. Multidisciplinary approaches combined with new technologies may help to uncover new microbial PUFA sources that may have nutraceutical and biotechnological importance.
... As for fungi, genera belonging to Mucoromycota (including Mucor, Rhizopus, Umbelopsis, Lichtheimia, Cunninghamella, and Mortierella) are well known for their ability to synthesize PUFAs, especially gamma-linolenic acid (GLA), which is of great pharmaceutical interest due to its anticancer properties, while it has been used to improve premenstrual tension and various skin diseases [13,17]. Especially, Cunninghamella echinulata is an important GLA producer [13,19,23], while Umbelopsis isabellina is regarded as a promising SCO producer, being able to accumulate lipids in high percentages, though less rich in GLA [16,[24][25][26]. ...
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.
... Chlorophyceae (green algae) is known as one of the most popular producers of long-chain PUFAs, such as alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). [127][128][129][130][131] Cryptocodinium cohnii, Prorocentrum triestinum, and Amphidinium sp. are the efficient DHA-producers, 129,132 while N. salina synthesize EPA at more than 25% of total lipids. 127 Around 1 kg biomass of Chlorella sp. ...
... [127][128][129][130][131] Cryptocodinium cohnii, Prorocentrum triestinum, and Amphidinium sp. are the efficient DHA-producers, 129,132 while N. salina synthesize EPA at more than 25% of total lipids. 127 Around 1 kg biomass of Chlorella sp. synthesizes 12%-32% of the EPA from total PUFAs (18%-42%) present in algal lipids (11%-40% on dry basis/kg of biomass) (Fig. 3). ...
Microalgae have garnered significant interest among researchers and industries as a potential source of valuable products that can find applications in the food, feed, biofuels, cosmetics, pharmaceuticals, and nutraceuticals industries. Microalgae can remediate various waste streams. This review discusses the current trends in the valorization of algal biomass and proposes integrated biorefinery approaches to produce high‐value products by sustainable routes. In such processes, the waste streams can be used as a source of nutrients for the production of microalgal biomass, rich in high‐quality lipids, digestible proteins and bioactive compounds. The integrated approaches presented in this study offer cost‐effective and environment‐friendly processing techniques while expanding the bioproducts portfolio in the food and therapeutics sector. The simultaneous or sequential extraction of valuable compounds from microalgae makes the biorefinery proposition appealing for waste minimization, revenue diversification, and complete feedstock utilization. The systematic exploration of microalgae on a large scale is required to evaluate its future as a favorable resource for biorefinery products. This would also necessitate detailed techno‐economic, environmental, and lifecycle assessment for the biorefinery processes. © 2021 Society of Chemical Industry and John Wiley & Sons, Ltd
... For all these reasons, glycerol overproduction and disposal may cause important environmental problems in the near future. The last years therefore, one of the most important topics that have been developed in both the Chemical and Microbial Technology, refers to the utilization of glycerol as starting or supplementary material that would be converted into various final added-value compounds [3,[8][9][10][11]. ...
... In agreement with the literature [2,12,70] the principal FAs found in variable quantities were mainly the oleate ( Δ9 C18:1), the palmitate (C16:0) and the linoleate ( Δ9,12 C18:2) ( Table 9). Poly-unsaturated FAs presenting double bonds ≥3 were not detected in high concentrations, since these compounds are the principal storage lipophilic compounds in oleaginous fungi and algae [2,8,33], and, in general, they can be produced in significant quantities inside the yeast cells only after appropriate genetic modifications [33]. Interestingly, R. kratochvilovae FMCC Y71 produced cellular lipids that contained high concentrations of Δ9,12 C18:2, in contrast to recent literature reports [99,100]. ...
Four non-conventional yeast strains belonging to the genera Yarrowia and Rhodosporidium were cultivated under nitrogen-limited conditions in shake flasks with biodiesel-derived glycerol employed as sole substrate. The strains R. toruloides DSM 4444 and Y. lipolytica ACA-DC 5033 displayed better performances and were further studied. At initial glycerol (Gly0) ≈50 g/L, the strain ACA-DC 5033 produced dry cell weight (DCW) ≈7.0 g/L, and also secondary extra-cellular metabolites as citric acid (CA) at ≈16 g/L and polyols (Pol) (mannitol, erythritol and arabitol) at ≈21 g/L. In double-media (besides nitrogen also Mg was limiting) for Gly0≈50 g/L, a shift towards CA production occurred (CAmax≈33 g/L). The strain DSM 4444 at Gly0≈50 g/L produced DCW=18.1 g/L containing lipids =30.3% in DCW. In single nitrogen-limited media with Gly0≈90 g/L, the strain ACA-DC 5033 produced Pol≈48 g/L and CA≈20 g/L, while the strain DSM 4444 produced DCW=27.3 g/L containing lipid =54.5% w/w. At the late growth phases, metabolites were re-consumed. Balanced growth phase (trophophase) and phase of secondary metabolite synthesis (idiophase) were successfully simulated with the aid of a modified Velhlust-Aggelis model. Lipid extraction process was studied for these two strains by using two different extraction methods. Yeast lipids contained mostly oleic acid, constituting suitable precursors for the synthesis of 2nd generation biodiesel.
