“…3). Similar results have already been reported for Y. lipolytica [44] and for the oleaginous yeast Lipomyces starkeyi [45]. As we found here, in both prior studies, a C/N of 35 led to higher accumulation of intracellular lipids.Fig.…”
Section: Resultssupporting
confidence: 93%
“…However, the bioreactor yielded higher lipid content than the BioLector did. This could be because the flowerplate of the BioLector is designed to obtain a high oxygen transfer rate [50], and it has been demonstrated that higher amounts of dissolved oxygen increase citric acid production at the expensive of lipid synthesis [43, 44]. Overall, the lipid content determined during the comparison of the two fermentation instruments allowed the same strains to be discriminated for lipid accumulation.…”
BackgroundBecause the model yeast Yarrowia lipolytica can synthesize and store lipids in quantities up to 20 % of its dry weight, it is a promising microorganism for oil production at an industrial scale. Typically, optimization of the lipid production process is performed in the laboratory and later scaled up for industrial production. However, the scale-up process can be complicated by genetic modifications that are optimized for one set of growing conditions can confer a less-than-optimal phenotype in a different environment. To address this issue, small cultivation systems have been developed that mimic the conditions in benchtop bioreactors. In this work, we used one such microbioreactor system, the BioLector, to develop high-throughput fermentation procedures that optimize growth and lipid accumulation in Y. lipolytica. Using this system, we were able to monitor lipid and biomass production in real time throughout the culture duration.ResultsThe BioLector can monitor the growth of Y. lipolytica in real time by evaluating scattered light; this produced accurate measurements until cultures reached an equivalent of OD600nm = 115 and a cell dry weight of 100 g L−1. In addition, a lipid-specific fluorescent probe was applied which reliably monitored lipid production up to a concentration of 12 g L−1. Through screening various growing conditions, we determined that a carbon/nitrogen ratio of 35 was the most efficient for lipid production. Further screening showed that ammonium chloride and glycerol were the most valuable nitrogen and carbon sources, respectively, for growth and lipid production. Moreover, a carbon concentration above 1 M appeared to impair growth and lipid accumulation. Finally, we used these optimized conditions to screen engineered strains of Y. lipolytica with high lipid-accumulation capability. The growth and lipid content of the strains cultivated in the BioLector were compared to those grown in benchtop bioreactors.ConclusionTo our knowledge, this is the first time that the BioLector has been used to track lipid production in real time and to monitor the growth of Y. lipolytica. The present study also showed the efficacy of the BioLector in screening growing conditions and engineered strains prior to scale-up. The method described here could be applied to other oleaginous microorganisms.
“…3). Similar results have already been reported for Y. lipolytica [44] and for the oleaginous yeast Lipomyces starkeyi [45]. As we found here, in both prior studies, a C/N of 35 led to higher accumulation of intracellular lipids.Fig.…”
Section: Resultssupporting
confidence: 93%
“…However, the bioreactor yielded higher lipid content than the BioLector did. This could be because the flowerplate of the BioLector is designed to obtain a high oxygen transfer rate [50], and it has been demonstrated that higher amounts of dissolved oxygen increase citric acid production at the expensive of lipid synthesis [43, 44]. Overall, the lipid content determined during the comparison of the two fermentation instruments allowed the same strains to be discriminated for lipid accumulation.…”
BackgroundBecause the model yeast Yarrowia lipolytica can synthesize and store lipids in quantities up to 20 % of its dry weight, it is a promising microorganism for oil production at an industrial scale. Typically, optimization of the lipid production process is performed in the laboratory and later scaled up for industrial production. However, the scale-up process can be complicated by genetic modifications that are optimized for one set of growing conditions can confer a less-than-optimal phenotype in a different environment. To address this issue, small cultivation systems have been developed that mimic the conditions in benchtop bioreactors. In this work, we used one such microbioreactor system, the BioLector, to develop high-throughput fermentation procedures that optimize growth and lipid accumulation in Y. lipolytica. Using this system, we were able to monitor lipid and biomass production in real time throughout the culture duration.ResultsThe BioLector can monitor the growth of Y. lipolytica in real time by evaluating scattered light; this produced accurate measurements until cultures reached an equivalent of OD600nm = 115 and a cell dry weight of 100 g L−1. In addition, a lipid-specific fluorescent probe was applied which reliably monitored lipid production up to a concentration of 12 g L−1. Through screening various growing conditions, we determined that a carbon/nitrogen ratio of 35 was the most efficient for lipid production. Further screening showed that ammonium chloride and glycerol were the most valuable nitrogen and carbon sources, respectively, for growth and lipid production. Moreover, a carbon concentration above 1 M appeared to impair growth and lipid accumulation. Finally, we used these optimized conditions to screen engineered strains of Y. lipolytica with high lipid-accumulation capability. The growth and lipid content of the strains cultivated in the BioLector were compared to those grown in benchtop bioreactors.ConclusionTo our knowledge, this is the first time that the BioLector has been used to track lipid production in real time and to monitor the growth of Y. lipolytica. The present study also showed the efficacy of the BioLector in screening growing conditions and engineered strains prior to scale-up. The method described here could be applied to other oleaginous microorganisms.
“…It is possible that Ilex paraguariensis has a significant presence of this lipid in its composition, which may explain the increase of this fatty acid in thigh meat of broilers fed with mate. Kolouchová et al (2015) reported that mono-unsaturated fatty acids reduce the alleged regulatory pool of intracellular free cholesterol, increasing LDL receptor activity and subsequently decreasing the blood circulating cholesterol concentrations. Palmitoleic and oleic acids rich diets have been reportedas able to improve blood circulating lipid profile, resulting in reduced total and LDL cholesterol (Griel et al 2008).…”
Yerba mate (Ilex paraguariensis)has in its composition organic compounds wich can modify chemical composition of broiler meat. This study aimed to evaluate the influence of yerba mate in chemical composition and lipid profile of broiler breast and thigh meat. The trial was conducted using 500 broiler chicks distributed in a completely randomized design with four treatments (0.1; 0.2; 0.4, and 0.6%) and five replicates with 25 chicks each. The chicken were raised form 1 to 42 days old and slaughtered for analysis of breast and thigh meat. The addition of mate did not affect the chemical composition and the lipid profile of the chicken breast, however, the composition of the meat of thigh meat was affected. It was observed effect of the addition of the yerba mate to saturated, unsaturated, monounsaturated and polyunsaturated fatty acids in the chicken meat. Stands out the reduction in saturated fatty acid concentrations and the increase in the concentrations of polyunsaturated fatty acids, especially Linolenic and Eicosapentaenoic, thus demonstrating the potential of mate grass in altering the lipid profile of poultry meat.
Mass spectrometry-based shotgun lipidomics was applied to the analysis of sphingolipids of 11 yeast strains belonging to four genera, that is Cryptococcus, Saccharomyces, Schizosaccharomyces, and Wickerhamomyces. The analysis yielded comprehensive results on both qualitative and quantitative representation of complex sphingolipids of three classes-phosphoinositol ceramide (PtdInsCer), mannosyl inositol phosphoceramide (MInsPCer), and mannosyl diinositol phosphoceramide (M(InsP) Cer). In total, nearly 150 molecular species of complex sphingolipids were identified. Individual strains were cultured at five different temperatures, that is 5, 10, 20, 30, and 40 °C (Wickerhamomyces genus only up to 30 °C), and the change in the culture temperature was found to affect the representation of both the sphingolipid classes and the length of the long-chain bases (LCB). Individual classes of sphingolipids differing in polar heads differed in the temperature response. The relative content of PtdInsCer increased with increasing temperature, whereas that of M(InsP) Cer decreased. Molecular species having C18-LCB were associated with low cultivation temperature, and a higher proportion of C20-LCB molecular species was produced at higher temperatures regardless of the type of polar head. On the other hand, the influence of temperature on the representation of very long-chain fatty acids (VLCFA) was less noticeable, the effect of the taxonomic affiliation of the strains being more pronounced than the cultivation temperature. For example, lignoceric and 2-hydrocylo-lignoceric acids were characteristic of the genera Cryptococcus and Schizosaccharomyces, and of Saccharomyces genus cultivated at high temperatures.
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