Effect of pH on the Content of Glycolipids in Aureobasidium pullulans S–l

When Aureobasidium pullulans was cultivated with initial pH 6.0 and pH 2.5 in shaking flasks, only a slight difference was observed in the content of lipids in cells. On the other hand, when cultured with initial pH 2.5 in a jar fermenter, the content of lipids in mycelia was larger than that with initial pH 6.0. The content of sugars in the lipids showed a similar tendency. Glucolipids were easily hydrolyzed with dilute acid into glucose, glucooligo- saccharide and lipids.     

Glucose as a growth medium factor regulating lipid composition of Yersinia pseudotuberculosis

Effects of glucose and growth temperature on Yersinia pseudotuberculosis O:1b serovar lipid composition have been studied. These growth parameters were shown to have drastic effects on biosynthetic processes in the pseudotuberculosis bacteria. The temperature effect is the most universal, extending to cell growth and to free lipid and lipopolysaccharide content and composition; it is most conspicuous in the bacteria cultivated on glucose-containing nutrient broth. The effect of glucose is selective, affecting only free lipids and depending on temperature (glucose favors phospholipid (PL) synthesis in the cold and inhibits it at 37°C); the effect of glucose is more evident in the cold. Determination of the contents of individual PL in percent dry bacterial weight indicates that the most obvious effect of glucose and/or growth temperature is on phosphatidylethanolamine (PE) content: on both media and at both temperatures an overall decrease in PL content stems from the inhibition of PE synthesis and is attended by decreasing ratio of neutral to acidic lipids.     

Effect of culture conditions on growth and production of docosahexaenoic acid (DHA) usingThraustochytrium aureum ATCC 34304

Environmental and medium factors were investigated as basic data for optimizing DHA production when usingThraustochytrium aureum. To study the effect of environmental conditions, the rotation speed and culture temperature were, changed. Plus the trend of the growth characteristics, lipid content in the biomass, and DHA content in lipids were evaluated according to various initial glucose concentrations. The biomass, lipid, and DHA analyses showed that the physiological characteristics ofT. aureum were closely related with the environmental and medium conditions, as in the case of other marine microorganisms. For example, a low rotation speed of 50 rpm lowered the cell growth rate as well as the DHA content in the lipids. A low temperature had a negative effect on the cell growth, yet a positive effect on the lipid content in the biomass. Different initial glucose concentrations had no effect on the lipid content in the biomass or DHA content in the lipids, yet did affect the cell growth. Accordingly, these results show that environmental and medium factors must be synthetically considered in order to optimize DHA production when usingT. aureum.     

Comparative lipid content ofCandida lipolytica grown on glucose and onn-hexadecane

Candida lipolytica, grown onn-hexadecane as the sole source of carbon and energy, contained 17.1% lipids in the logarithmic phase of growth, and 7.3% lipids in the stationary phase of growth. When the yeast was grown on glucose, it contained 6.2% lipids in the logarithmic phase of growth, and 3.6% lipids in the stationary phase of growth. Fatty acids, that could be extracted by petroleum ether after saponification, constituted the major part of the fatty acids ofC. lipolytica in its logarithmic phase of growth on glucose. They constituted only a minor amount of the fatty acids in the stationary phase of growth on glucose. The reverse was true when the yeast was grown onn-hexadecane. The broth contained more free, petroleum ether-soluble fatty acids when the cellular lipid content was high than when it was low. Overnight starvation ofC. lipolytica grown onn-hexadecane in a carbon-free nutrient medium, removed the residual cell-bound hydrocarbon, increased the cell population by one half and decreased the cellular lipid content (as % of dry yeast) by one third. Various methods for the determination of lipids, described as appropriate for yeasts were compared. The highest yields were obtained by extraction of the freeze-dried paste, at room temperature, with a 1:1 chloroform-methanol mixture.     

Changes of γ-linolenic acid content inRhizopus arrhizus duringl(+)-lactic acid fermentation

Effects of nitrogen source, temperature and pH onl(+)-lactic acid production and γ-linolenic acid (GLA) accumulation byRhizopus arrhizus were examined. The nitrogen source had a minor effect on lactate synthesis but influenced the total lipid content and the fatty acid composition in fungus. Higher temperature favorably influenced the rate of both lactic acid production and lipid formation in the biomass and caused a decrease in the yields of oligounsaturated fatty acids. At higher temperature and after glucose exhaustion, degradation of lactate increased. A low pH value negatively affected the formation of lipids and lactate synthesis. The highest value of GLA in the lipid (25.5%,W/W) was reached at the end of lactate synthesis, but maximum yields of total lipids were achieved when the cultivation continued in the presence of lactate until polyols were exhausted.     

A comparative study of carbon energy reserve metabolism ofC. tropicalis growing on glucose and on hydrocarbons

Summary Glycogen was markedly accumulated inCandida tropicalis growing on glucose with increasing limitation of external substrate supply. The same effect was caused by a N-free medium. The lipid content did not show any significant change in either case. On the hydrocarbon substrate, lipid increased as substrate availability decreased, whereas glycogen accumulation was only slight. However, the increase of lipid content on hydrocarbons did not reach the same level of accumulation as glycogen on glucose. In N-free medium both glycogen and lipids were accumulated, indicating that glycogen is not substituted by lipids as the carbon energy reserve on a hydrocarbon substrate.In addition, a refined shift technique is described. The disturbing influence of excess substrate at the beginning of a shift from glucose to hydrocarbons is avoided by a portioned substrate feeding according to the cell activity.     

葡萄糖对小球藻(Chloralla sp. HN08)光合作用和生长的影响

【目的】探讨葡萄糖作为外加碳源对热带海洋小球藻(Chloralla sp.HN08)生物质生产和脂、光合色素、碳水化合物及可溶性蛋白等细胞主要成份含量的影响。【方法】分析比较小球藻HN08在光合自养和兼养(添加10 g/L葡萄糖)2种营养方式下的生长速率、细胞密度、光合放氧速率、油脂相对含量,以及可溶性总糖、淀粉和可溶性蛋白的含量。【结果】结果表明,在光照条件下葡萄糖(10 g/L)能促进小球藻(Chloralla sp.HN08)生长,提高细胞终密度,而异养条件下藻细胞逐渐衰亡。兼养条件下,细胞相对生长速率及细胞终密度分别是自养条件下的6.8倍和1.3倍。兼养藻细胞中可溶性糖、淀粉、油脂含量显著高于(P0.05)光合自养细胞,然而可溶性蛋白质和光合色素含量显著低于(P0.05)光合自养细胞。添加葡萄糖的小球藻液的光饱和点和呼吸速率均高于光自养条件下的细胞,但2种培养条件下藻液的净光合速率无显著差异(P0.05)。【结论】光照条件下,添加葡萄糖可显著提高小球藻HN08相对生长速率和细胞终密度,促进油脂与淀粉的积累。     

Influence of selected sugars and temperature on fatty acids composition in Candida lipolytica

Summary An investigation was made of the effects of temperature and of mono- and disaccharides on lipids, biomass, odd-chain and unsaturated fatty acids production of Candida lipolytica. With different sugars as carbon source at 30°C, the order for biomass production was: fructose > glucose > sucrose > lactose > galactose, while lipids production/g biomass decreased as follows: lactose, sucrose, galactose, fructose and glucose. On the other hand, the odd-chain fatty acids contents decreased in the following order: fructose, lactose, glucose, sucrose and galactose. Lowering the temperature of cultivation to 15°C, biomass, lipids and unsaturated fatty acids decreased. However, a notable decrease in the content of odd-chain fatty acids was detected.     

Lipid composition of heteroploid cells in culture: effects of prednisolone

Prednisolone-induced alterations in the content and composition of the total lipids, cholesterol, phospholipids and fatty acids were studied in HeLa S3 cells. After relatively long exposure of the cell cultures to the hormone analog (24 to 72 hours), total cell lipid content was decreased. Partial inhibition of cholesterol and phospholipid metabolism resulted in a shift in the molar ratio of these two lipid constituents. Total fatty acid content was unaffected by prednisolone but there were minor changes in the relative distribution of the individual fatty acids. The observed decrease in cholesterol turnover after addition of prednisolone was primarily due to the reduced uptake of intact cholesterol from the culture medium. This was compensated, in part, by an increased de novo synthesis of cholesterol from acetate and glucose.     

Organic nitrogen of tomato waste hydrolysate enhances glucose uptake and lipid accumulation in Cunninghamella echinulata

Aims: To investigate the effect of organic nitrogen on lipogenesis during growth of Cunninghamella echinulata on tomato waste hydrolysate (TWH) media. Methods and Results: Cunninghamella echinulata grown on a TWH medium rapidly took up glucose and produced large amounts of lipids. However, when some quantities of the organic nitrogen were removed from TWH (by acid followed by alkaline precipitation of proteins) the uptake of glucose was dramatically reduced and large quantities of fungal biomass having low lipid content were produced. Nevertheless, when glycerol was used as carbon source instead of glucose, the uptake rate as well as the biomass production and the lipid accumulation processes were unaffected by the TWH organic nitrogen removal. Finally, when the fungus was grown on a glucose supplemented TWH medium that contained no assimilable organic nitrogen (after further precipitation of proteins with methanol), the produced biomass contained non-negligible quantities of lipids, although glucose uptake remained low. Lipid analysis showed that the produced lipids comprised mainly of neutral lipids, which were preferentially consumed during lipid turnover. Lipid production on the original TWH medium having glucose as carbon source was 0·48 g of lipid per gram of dry biomass, corresponding to 8·7 g of lipid per litre of growth medium. The produced lipids contained 11·7%γ-linolenic acid (GLA), hence the GLA yield was more than 1 g l⁻¹. Conclusions: Organic nitrogen compounds found in TWH favour glucose (but not glycerol) uptake and lipid accumulation in C. echinulata. Significance and Impact of the Study: Agro-industrial wastes containing organic nitrogen, such as tomato waste, are produced in vast amounts causing severe environmental problems. These wastes could be used as fermentation feedstock to produce microbial lipids.     

Evaluation ofin vivo insulin action and glucose metabolism in milk-fed rats

Milk diet has long been recommended in the management of gastrointestinal pathologies. Since milk feeding represents a high fat-low carbohydrate diet and it is acknowledged that insulin resistance is one of the consequences of high fat feeding, it is important to know whether or not chronic milk feeding leads to an impairment of the insulin-mediated glucose metabolism. To examine this question, adult female rats were given raw cow's milk (50% of total calories as lipids) for 18 days. They were compared to rats raised in parallel and fed the standard laboratory diet (15% of total calories as lipids). At the end of the 18 day period, body weight, daily caloric intake, basal plasma glucose and insulin levels in the milk-fed rats were similar to those in the control rats.In vivo insulin action was assessed with the euglycemichyperinsulinemic clamp technique in anesthetized animals. These studies were coupled with the 2-deoxyglucose technique allowing a measurement of glucose utilization by individual tissues. In the milk fed rats: 1) the basal rate of endogenous glucose production was significantly (p<0.01) reduced (by 20%); 2) their hepatic glucose production was however normally suppressed by hyperinsulinemia; 3) their basal glucose utilization rate was significantly (p<0.01) reduced (by 20%); 4) their glucose utilization rate by the whole-body mass or by individual tissues was normally increased by hyperinsulinemia. These results indicate that insulin action in adult rats is not grossly altered after chronic milk-feeding, at least under the present experimental conditions.     

Lipid Composition of Tomato Leaves as Related to Plant Cold Tolerance

We studied the effects of a low nonlethal temperature (6°C) on the content and composition of polar lipids and their fatty acids in tomato (Lycopersicon esculentum Mill., cv. Sibirskie skorospelye) leaves. We demonstrated that chilling resulted in a decrease in the content of total polar lipids per 1 mg protein. The content of lipids in chloroplast membranes (monogalactosyldiacylglycerols, digalactosyldiacylglycerols, sulfoquinovosyldiacylglycerols, and phosphatidylglycerols) changed less substantially than the content of phospholipids in other cell organelles and in the cytoplasm. Neutral lipids comprised only 1% of total lipids, and their content also decreased after chilling. The relative amounts of unsaturated and saturated fatty acids in polar lipids were practically unchanged. The conclusion was drawn that the maintenance of a high level of chloroplast membrane lipids under low temperatures could play an important role in the survival of cold-tolerant plants.     

Annual cycles of growth rate, feeding rate, food conversion, plasma glucose and plasma lipids in a population of European sea bass (Dicentrarchus labrax L.) farmed in floating marine cages

Specific growth rate, average daily food intake, gross conversion efficiency, plasma total lipids, triglycerides, total cholesterol and glucose were evaluated annually in a population of European sea bass (Dicentrarchus labrax L.) intensively reared in floating marine cages. These data are aimed at improving productivity and allows more precise limits to be placed on the hematological parameters which are a necessary precondition for the assessment of fish condition and health. Specific growth rate (%), average daily food intake (%) and gross conversion efficiency showed relatively higher values in the warm spring and summer months. The specific growth rate correlated better with the seawater temperature and the average daily food intake than with the photoperiod. The concentrations of plasma total lipids, triglycerides and glucose were higher in summer. Plasma total cholesterol concentrations were higher during the pregametogenesis and gametogenesis periods. Plasma triglycerides correlated better with photoperiod and plasma glucose than with the seawater temperature.     

Intracellular accumulation of mannosylerythritol lipids as storage materials by Candida antarctica

Summary Candida antarctica strain T-34, which was isolated as a biosurfactant producer, was found to produce organic acids and polyols extracellularly but not to produce biosurfactants, when grown on glucose or other carbohydrates as the sole carbon source. It was also observed microscopically that the strain contained oil globules within the cells. The intracellular lipids of the strain mainly consisted of triglycerides and mannosylerythritol lipids (MEL). The MEL content of the cells during the culture exceeded 10% of the dry cell weight, and the pattern of variation of the MEL content was very similar to that of triglycerides. All three stock strains of C. antarctica tested also accumulated a relatively large amount of MEL from glucose. These results suggested that these strains accumulated the MEL intracellularly as one of the storage materials together with triglycerides.Offprint requests to: D. Kitamoto     

Zur mixotrophen Kultivation von Chlorella vulgaris in homokontinuerlicher Chemostatkultur

Cells of Chlorella vulgaris, BEIJ. Greifswald 9, were grown on autotrophic and mixotrophic conditions using glucose and acetate as organic substrates. It was shown that these C-sources applicated in a suitable range of concentrations increase the growth rate and the productivity of the algal cultures. The cells grown on mixotrophic conditions have a higher total pigment content and exhibit variations in the ratio chlorophyll a/chlorophyll b. In addition the contents of proteins, lipids, carbohydrates, and nucleic acids of the biomass were shown to be dependent on the kind of the organic substrate used.     

Effect of glucose on the biochemical properties of the bacterial cytoplasmic membrane

The cytoplasmic membrane isolated from cells ofCitrobacter freundii growing in a cultivation medium with glucose was found to exhibit a decreased ability to oxidize formate and succinate and a decreased activity of formate, succinate and lactate dehydrogenase as compared with the cytoplasmic membrane of cells growing on galactose. The activation energy for dehydrogenation and oxidation of succinate simultaneously increases. The quantitative content of cytochromes and quinones in both types of membranes does not differ considerably. The two types of membranes also do not mutually differ in the qualitative and quantitative representation of fatty acids in lipids and in the sensitivity to the effect of low concentrations of detergents. The mass ratio of proteins and lipids is lower in the membranes of cells grown on glucose.     

Production of docosahexaenoic acid by Thraustochytrium aureum

Summary Thraustochytrium aureum ATCC 34 304 contained approximately 50% of total fatty acids as docosahexaenoic acid (DHA). Lipid content of the biomass was dramatically influenced by medium composition and ranged from 0.3 to 16% of the biomass weight. Increasing the culture medium glucose concentration from 5 to 20 g/l caused the lipid content of the biomass to increase from 2.7 to 16.5% and the DHA yield to increase from 26 to 270 mg/l of whole culture broth. The proportion of DHA in lipids was independent of glucose concentration. The fatty acid profile observed in the high-yielding DHA culture was: 16:0, 19.2%; 18:1, 9.8%; 18:2, 2.4%; 20:4, 4.9%; 20:5, 3.6%; 22:6, 48.5% and others, 7.1%. For DHA production, fungal cultures were incubated on an orbital shaker under light at 25° C for 6 days. Offprint requests to: O. P. Ward     

Novel sophorose lipids from microbial conversion of 2-alkanols

Summary Using glucose/rac. 2-alkanols as carbon sources for the cultivation of Candida bombicola novel types of sophorose lipids were isolated as major products. Among all lipophilic constituents 2-alkanol was found as the major hydrophobic moiety (> 75%) involved in three new sophorose lipids differing only in the degree of acetylation of the sophorose. Additional monooxygenation of the alcohol leads to the incorporation of 2,(-1)-alkandiol; glycolipids with up to four glucose units were obtained.     

Dynamic flux balance analysis of biomass and lipid production by Antarctic thraustochytrid Oblongichytrium sp. RT2316-13

Production of biomass and lipids in batch cultures of the Antarctic thraustochytrid Oblongichytrium sp. RT2316-13, is reported. The microorganism proved capable of producing nearly 67% docosahexaenoic acid (DHA) and 15% eicosapentaenoic acid (EPA) in its total lipid fraction. Biomass with a maximum total lipid content of 33.5% (wt/wt) could be produced at 15°C in batch culture using a medium containing glucose (20 g/L), yeast extract (10.5 g/L), and other minor components. A lower culture temperature (5°C) reduced biomass and lipid productivities compared to culture at 15°C, but enhanced the DHA and EPA content of the lipids by 6.4- and 3.3-fold, respectively. Both a simple minimally structured mathematical model and a more complex genome-scale metabolic model (GEM) allowed the fermentation profiles in batch cultures to be satisfactorily simulated, but the GEM provided much greater insight in the biochemical and physiological phenomena underlying the observed behavior. Unlike the simpler model, the GEM could be interrogated for the possible effects of various external factors such as oxygen supply, on the expected outcomes. In silico predictions of oxygen effects were consistent with literature observations for DHA producing thraustochytrids.     

Production of arachidonic acid byMortierella alpina ATCC 32222

Summary WhenMortierella alpina ATCC 32222 was incubated in a glucose salts medium at 25°C the biomass (17.5 g/l) contained 9.62% arachidonic acid which amounted to 54% (w/w) of total biomass lipids. When the glucose concentration in the medium was varied from 0 to 150 g/l, the percentage of arachidonic acid in biomass and in lipids was highest at a glucose concentration of 30 g/l, but highest yield of arachidonic acid per litre of culture broth was observed at a glucose concentration of 100 g/l. While production of biomass reached a plateau of 17 g/l after a 3-day incubation at 25°C, the percentage of arachidonic acid in lipids and biomass increased dramatically from 3 to 6 days with a concurrent arachidonic acid yield increase from 0.89 to 1.63 g/l. Optimum initial culture pH for arachidonic acid production was in the range 6.0–6.7. By increasing the concentration of the glucose salts medium three-fold, yields of biomass and arachidonic acid were increased to 35.8 g/l and 3.73 g/l, respectively.