Metabolism of Storage Lipids and the Role of Lipid Droplets in the Yeast Schizosaccharomyces pombe

Storage lipids, triacylglycerols (TAG), and steryl esters (SE), are predominant constituents of lipid droplets (LD) in fungi. In several yeast species, metabolism of TAG and SE is linked to various cellular processes, including cell division, sporulation, apoptosis, response to stress, and lipotoxicity. In addition, TAG are an important source for the generation of value-added lipids for industrial and biomedical applications. The fission yeast Schizosaccharomyces pombe is a widely used unicellular eukaryotic model organism. It is a powerful tractable system used to study various aspects of eukaryotic cellular and molecular biology. However, the knowledge of S. pombe neutral lipids metabolism is quite limited. In this review, we summarize and discuss the current knowledge of the homeostasis of storage lipids and of the role of LD in the fission yeast S. pombe with the aim to stimulate research of lipid metabolism and its connection with other essential cellular processes. We also discuss the advantages and disadvantages of fission yeast in lipid biotechnology and recent achievements in the use of S. pombe in the biotechnological production of valuable lipid compounds.     

Solvent-Aided Crystallization for Biodiesel Purification

The application of solvent-aided crystallization (SAC) is based on the addition of a solvent, here 1-butanol, to crude biodiesel to catalyze the purification process by separating biodiesel from contaminants via crystallization process. Response surface methodology was applied to optimize the process parameters of SAC, represented by biodiesel purity. The purified biodiesel was analyzed by means of gas chromatography-mass spectrometry for the composition of the present fatty acid methyl ester (FAME). Under the predicted optimum process conditions within the experimental ranges for the highest biodiesel purity, the predicted biodiesel purity was 99.375 %.     

“Arbequina” Olive Oil Composition Is Affected by the Application of Regulated Deficit Irrigation during Pit Hardening Stage

Three new regulated deficit irrigation (RDI) treatments were applied to “Arbequina” olive orchards during pit hardening. Oil quality was determined by measuring analytical parameters for olive oil grading, antioxidant activity, total phenol content, fatty acid profile, volatile compounds profile, and sensory analysis. Oils from RDI were classified as “extra virgin olive oil” and their quality was improved due to their higher antioxidant potential (ABTS⁺ [increased ~75%] and DPPH^˙ [increased ~25%] assays) and phenols (increased ~53%) than control. Concentration of total volatile compounds decreased (~27%) but RDI olive oils showed a more balanced profile (alcohols, aldehydes, and esters). Monounsaturated fatty acid content increased (~5%) and atherogenic and thrombogenic indexes decreased (~8.5%) in RDI olive oil. Regarding sensory analysis, RDI provided more balanced oils with higher fruit aroma than control. Other benefits of RDI olive oil, when compared with oil from full irrigated orchards are reduced use of water and improved functional and sensory quality.     

多铜氧化酶在大肠杆菌中的分泌表达

生物胺是存在于发酵食品中的一类有机物,过量摄入会危害人体健康。多铜氧化酶中的某些酶具有降解多种生物胺的活性,在减控发酵食品中的氨(胺)类危害物方面具有良好的应用前景。研究多铜氧化酶的分泌表达,对酶的特性改造和工业化生产与应用具有重要意义。本研究通过在解淀粉芽孢杆菌来源的多铜氧化酶N端融合信号肽PhoA实现了多铜氧化酶在大肠杆菌中的分泌表达,胞外酶活为69.8 U/L。通过优化诱导条件和酶的分泌确定了多铜氧化酶最优发酵条件为诱导温度25℃、IPTG浓度0.05 mmol/L、诱导时菌体OD600=1.0、诱导6 h后添加150 mmol/L甘氨酸;发酵40 h时胞外多铜氧化酶酶活达到238.1 U/L,是优化前的3.4倍。     

Influence of Serum and Hypoxia on Incorporation of [14C]-d-Glucose or [14C]-l-Glutamine into Lipids and Lactate in Murine Glioblastoma Cells

Glucose and glutamine are essential energy metabolites for brain tumor growth and survival under both normoxic and hypoxic conditions. Both metabolites can contribute their carbons to lipid biosynthesis. We used uniformly labeled [¹⁴C]‐U‐d ‐glucose and [¹⁴C]‐U‐l ‐glutamine to examine the profile of de novo lipid biosynthesis in the VM‐M3 murine glioblastoma cells. The major lipids synthesized included phosphatidylcholine (PtdCho), phosphatidylethanolamine (EtnGpl), phosphatidylinositol (PtdIns), phosphatidylserine (PtdSer), sphingomyelin (CerP Cho), bis(monoacylglycero)phosphate (BMP)/phosphatidic acid (PtdOH), cholesterol (C), cardiolipin (Ptd₂Gro), and gangliosides. Endogenous lipid synthesis, using either glucose or glutamine, was greater in media without fetal bovine serum (FBS) than in media containing 10 % FBS under normoxia. De novo lipid synthesis was greater using glucose carbons than glutamine carbons under normoxia. The reverse was observed for most lipids under hypoxia suggesting an attenuation of glucose entering the TCA cycle. Lactate was produced largely from glucose carbons with minimal lactate derived from glutamine under either normoxia or hypoxia. Accumulation of triacylglycerols (TAG), containing mostly saturated and mono‐unsaturated fatty acids, was observed under hypoxia using carbons from either glucose or glutamine. The data show that the incorporation of labeled glucose and glutamine into most synthesized lipids was dependent on the type of growth environment, and that the VM‐M3 glioblastoma cells could acquire lipids, especially cholesterol, from the external environment for growth and proliferation.     

Structurally Diverse Polyamines: Solid‐Phase Synthesis and Interaction with DNA

A versatile solid‐phase approach based on peptide chemistry was used to construct four classes of structurally diverse polyamines with modified backbones: linear, partially constrained, branched, and cyclic. Their effects on DNA duplex stability and structure were examined. The polyamines showed distinct activities, thus highlighting the importance of polyamine backbone structure. Interestingly, the rank order of polyamine ability for DNA compaction was different to that for their effects on circular dichroism and melting temperature, thus indicating that these polyamines have distinct effects on secondary and higher‐order structures of DNA.     

High-Fat Diet Alters Serum Fatty Acid Profiles in Obesity Prone Rats: Implications for In Vitro Studies

High‐fat diets (HFD) are commonly used in rodents to induce obesity, increase serum fatty acids and induce lipotoxicity in various organs. Invitro studies commonly utilize individual free fatty acids (FFA) to study lipid exposure in an effort to model what is occurring in vivo; however, these approaches are not physiological as tissues are exposed to multiple fatty acids in vivo. Here we characterize circulating lipids in obesity‐prone rats fed an HFD in both fasted and fed states with the goal of developing physiologically relevant fatty acid mixtures for subsequent in vitro studies. Rats were fed an HFD (60 % kcal fat) or a control diet (10 % kcal fat) for 3 weeks; liver tissue and both portal and systemic blood were collected. Fatty acid profiles and absolute concentrations of triglycerides (TAG) and FFA in the serum and TAG, diacylglycerol (DAG) and phospholipids in the liver were measured. Surprisingly, both systemic and portal serum TAG were ~40 % lower in HFD‐fed compared to controls. Overall, compared to the control diet, HFD feeding consistently induced an increase in the proportion of circulating polyunsaturated fatty acids (PUFA) with a concomitant decline in monounsaturated fatty acids (MUFA) and saturated fatty acids (SFA) in both serum TAG and FFA. The elevations of PUFA were mostly attributed to increases in n‐6 PUFA, linoleic acid and arachidonic acid. In conclusion, fatty acid mixtures enriched with linoleic and arachidonic acid in addition to SFA and MUFA should be utilized for in vitro studies attempting to model lipid exposures that occur during in vivo HFD conditions.     

FAME Production and Fatty Acid Profiles from Moist Chlorella sp. and Nannochloropsis oculata Biomass

In the present study, we investigated the production of fatty acid methyl esters (FAME) from moist Chlorella sp. and Nannochloropsis oculata biomass using a hydrolysis–esterification process. Additionally, we evaluated for the first time the fatty acid profile before and after this process. Hydrolysis of the lipid fraction was performed on a moist biomass in the presence of differing amounts of an acid catalyst in both 50 and 100 % w/w water relative to the biomass. The esterification of the crude extracts of the free fatty acids (FFA) was then investigated. The experiments show that in the presence of 50 % w/w water relative to the biomass, the hydrolysis–esterification process results in higher FFA and FAME yields. The analysis of the fatty ester profiles did not reveal any degradation of the FFA from the microalgae biomass under the hydrolysis–esterification conditions. The results were compared with both extraction–transesterification and direct transesterification processes using dry biomass. The extraction–transesterification and hydrolysis–esterification processes resulted in similar FAME yields and similar profiles of the fatty esters from dry and moist biomass materials, respectively.     

Controlling Argan Seed Quality by NIR

The suitability of using visible/near infrared spectroscopy (Vis/NIR), as a rapid and non‐destructive technique for monitoring the quality of argan seeds (Argania spinosa Skeels) was studied. The analyzed parameters were the fatty acid composition of argan seed oil, seed moisture content, seed oil content and oil stability index (OSI). The ratio between major unsaturated and saturated fatty acids (U/S) during the oxidation assay at constant temperature was studied. Values from infrared drying were used as a laboratory reference for the moisture. Argan seed oil content was determined by Soxhlet extraction. A fatty acid analysis was carried out by gas chromatography and the OSI was determined by the Rancimat test. Predictive models of argan seed moisture, ratio U/S and OSI showed good accuracy. Therefore, Vis/NIR measurements can be used for controlling several argan seed quality parameters. This procedure might be of interest to the argan oil industry, which is currently in the process of modernization and expansion.     

Chemical Composition of Seed Oils Recovered from Different Pear (Pyrus communis L.) Cultivars

Lipophilic bioactive compounds in oils recovered from the seeds of eight pear (Pyrus communis L.) cultivars were studied. Oil yield in pear seeds ranged between 16.3 and 31.5 % (w/w) dw. The main fatty acids were palmitic acid (6.13–8.52 %), oleic acid (27.39–38.17 %) and linoleic acid (50.73–63.78 %), all three representing 96–99 % of the total detected fatty acids. The range of total tocochromanols was between 120.5 and 216.1 mg/100 g of oil. Independent of the cultivar, the γ‐tocopherol was the main tocochromanol and constituted approximately 88 %. The contents of the carotenoids and squalene were between 0.69–2.99 and 25.5–40.8 mg/100 g of oil, respectively. The β‐sitosterol constituted 83.4–87.6 % of total sterols contents, which ranged between 276.4 and 600.1 mg/100 g of oil. Three significant correlations were found between oil yield and total contents of sterols (r = ?0.893), tocochromanols (r = ?0.955) and carotenoids (r = ?0.685) in pear seed oils.