ARTP Mutagenesis of Schizochytrium sp. PKU#Mn4 and Clethodim-Based Mutant Screening for Enhanced Docosahexaenoic Acid Accumulation

Schizochytrium species are one of the best oleaginous thraustochytrids for high-yield production of docosahexaenoic acid (DHA, 22:6). However, the DHA yields from most wild-type (WT) strains of Schizochytrium are unsatisfactory for large-scale production. In this study, we applied the atmospheric and room-temperature plasma (ARTP) tool to obtain the mutant library of a previously isolated strain of Schizochytrium (i.e., PKU#Mn4). Two rounds of ARTP mutagenesis coupled with the acetyl-CoA carboxylase (ACCase) inhibitor (clethodim)-based screening yielded the mutant A78 that not only displayed better growth, glucose uptake and ACCase activity, but also increased (54.1%) DHA content than that of the WT strain. Subsequent optimization of medium components and supplementation improved the DHA content by 75.5 and 37.2%, respectively, compared with that of mutant A78 cultivated in the unoptimized medium. Interestingly, the ACCase activity of mutant A78 in a medium supplemented with biotin, citric acid or sodium citrate was significantly greater than that in a medium without supplementation. This study provides an effective bioengineering approach for improving the DHA accumulation in oleaginous microbes.     

Docosahexaenoic Acid Is Naturally Concentrated at the sn-2 Position in Triacylglycerols of the Australian Thraustochytrid Aurantiochytrium sp. Strain TC 20

The Labyrinthulomycetes or Labyrinthulea are a class of protists that produce a network of filaments that enable the cells to glide along and absorb nutrients. One of the main two Labyrinthulea groups is the thraustochytrids, which are becoming an increasingly recognised and commercially used alternate source of long-chain (LC, ≥C₂₀) omega-3 containing oils. This study demonstrates, to our knowledge for the first time, the regiospecificity of the triacylglycerol (TAG) fraction derived from Australian thraustochytrid Aurantiochytrium sp. strain TC 20 obtained using ¹³C nuclear magnetic resonance spectroscopy (¹³C NMR) analysis. The DHA present in the TC 20 TAG fraction was determined to be concentrated in the sn-2 position, with TAG (16:0/22:6/16:0) identified as the main species present. The sn-2 preference is similar to that found in salmon and tuna oil, and differs to seal oil containing largely sn-1,3 LC-PUFA. A higher concentration of sn-2 DHA occurred in the thraustochytrid TC 20 oil compared to that of tuna oil.     

Biosynthesis of Polyunsaturated Fatty Acids in Marine Invertebrates: Recent Advances in Molecular Mechanisms

Virtually all polyunsaturated fatty acids (PUFA) originate from primary producers but can be modified by bioconversions as they pass up the food chain in a process termed trophic upgrading. Therefore, although the main primary producers of PUFA in the marine environment are microalgae, higher trophic levels have metabolic pathways that can produce novel and unique PUFA. However, little is known about the pathways of PUFA biosynthesis and metabolism in the levels between primary producers and fish that are largely filled by invertebrates. It has become increasingly apparent that, in addition to trophic upgrading, de novo synthesis of PUFA is possible in some lower animals. The unequivocal identification of PUFA biosynthetic pathways in many invertebrates is complicated by the presence of other organisms within them. These organisms include bacteria and algae with PUFA biosynthesis pathways, and range from intestinal flora to symbiotic relationships that can involve PUFA translocation to host organisms. This emphasizes the importance of studying biosynthetic pathways at a molecular level, and the continual expansion of genomic resources and advances in molecular analysis is facilitating this. The present paper highlights recent research into the molecular and biochemical mechanisms of PUFA biosynthesis in marine invertebrates, particularly focusing on cephalopod molluscs.     

Alternative Sources of n-3 Long-Chain Polyunsaturated Fatty Acids in Marine Microalgae

The main source of n-3 long-chain polyunsaturated fatty acids (LC-PUFA) in human nutrition is currently seafood, especially oily fish. Nonetheless, due to cultural or individual preferences, convenience, geographic location, or awareness of risks associated to fatty fish consumption, the intake of fatty fish is far from supplying the recommended dietary levels. The end result observed in most western countries is not only a low supply of n-3 LC-PUFA, but also an unbalance towards the intake of n-6 fatty acids, resulting mostly from the consumption of vegetable oils. Awareness of the benefits of LC-PUFA in human health has led to the use of fish oils as food supplements. However, there is a need to explore alternatives sources of LC-PUFA, especially those of microbial origin. Microalgae species with potential to accumulate lipids in high amounts and to present elevated levels of n-3 LC-PUFA are known in marine phytoplankton. This review focuses on sources of n-3 LC-PUFA, namely eicosapentaenoic and docosahexaenoic acids, in marine microalgae, as alternatives to fish oils. Based on current literature, examples of marketed products and potentially new species for commercial exploitation are presented.     

Anxiolytic-Like Effect of a Salmon Phospholipopeptidic Complex Composed of Polyunsaturated Fatty Acids and Bioactive Peptides

A phospholipopeptidic complex obtained by the enzymatic hydrolysis of salmon heads in green conditions; exert anxiolytic-like effects in a time and dose-dependent manner, with no affection of locomotor activity. This study focused on the physico-chemical properties of the lipidic and peptidic fractions from this natural product. The characterization of mineral composition, amino acid and fatty acids was carried out. Stability of nanoemulsions allowed us to realize a behavioral study conducted with four different tests on 80 mice. This work highlighted the dose dependent effects of the natural complex and its various fractions over a period of 14 days compared to a conventional anxiolytic. The intracellular redox status of neural cells was evaluated in order to determine the free radicals scavenging potential of these products in the central nervous system (CNS), after mice sacrifice. The complex peptidic fraction showed a strong scavenging property and similar results were found for the complex as well as its lipidic fraction. For the first time, the results of this study showed the anxiolytic-like and neuroprotective properties of a phospholipopeptidic complex extracted from salmon head. The applications on anxiety disorders might be relevant, depending on the doses, the fraction used and the chronicity of the supplementation.     

Culturable Diversity of Thraustochytrids from Coastal Waters of Qingdao and Their Fatty Acids

Thraustochytrids have gained significant attention in recent years because of their considerable ecological and biotechnological importance. Yet, the influence of seasons and habitats on their culturable diversity and lipid profile remains poorly described. In this study, a total of 58 thraustochytrid strains were isolated from the coastal waters of Qingdao, China. These strains were phylogenetically close to five thraustochytrid genera, namely Botryochytrium, Oblongichytrium, Schizochytrium, Thraustochytrium, and Sicyoidochytrium. Most of the isolated strains were classified into the genera Thraustochytrium and Oblongichytrium. Further diversity analysis revealed that samples collected from nutrient-rich habitats and during summer/fall yielded significantly higher culturable diversity of thraustochytrids than those from low-nutrient habitats and winter/spring. Moreover, sampling habitats and seasons significantly impacted the fatty acid profiles of the strains. Particularly, the Oblongichytrium sp. OC931 strain produced a significant amount (153.99 mg/L) of eicosapentaenoic acid (EPA), accounting for 9.12% of the total fatty acids, which was significantly higher than that of the previously reported Aurantiochytrium strains. Overall, the results of this study fill the gap in our current understanding of the culturable diversity of thraustochytrids in the coastal waters and the impact of the sampling habitats and seasons on their capacity for lipid accumulation.     

花生籽仁发育过程中脂肪酸积累规律研究

以高O／L花生品种8130为材料对其籽仁发育过程中脂肪酸的累积过程进行分析。结果显示,花生籽仁中,含有8种主要的脂肪酸,按含量高低依次为油酸、亚油酸、棕榈酸、硬脂酸、山嵛酸、花生酸、二十四烷酸、花生烯酸;花生籽仁发育过程中,油酸含量逐渐增多,其它脂肪酸呈逐渐降低的变化规律。饱满籽仁中亚油酸和油酸含量占全部脂肪酸的80％以上。脂肪酸总量在籽仁发育前期增加迅速,然后缓慢增长,到籽仁成熟期又迅速增加,在收获期达到峰值,油酸亚油酸比值（O／L）,随籽仁发育呈逐渐增加的变化规律,饱满籽仁达到最大。     

Investigation of Deep-Sea Ecosystems Using Marker Fatty Acids: Sources of Essential Polyunsaturated Fatty Acids in Abyssal Megafauna

Abyssal seafloor ecosystems cover more than 50% of the Earth’s surface. Being formed by mainly heterotrophic organisms, they depend on the flux of particulate organic matter (POM) photosynthetically produced in the surface layer of the ocean. As dead phytoplankton sinks from the euphotic to the abyssal zone, the trophic value of POM and the concentration of essential polyunsaturated fatty acids (PUFA) decrease. This results in pronounced food periodicity and limitations for bottom dwellers. Deep-sea invertebrate seston eaters and surface deposit feeders consume the sinking POM. Other invertebrates utilize different food items that have undergone a trophic upgrade, with PUFA synthesized from saturated and monounsaturated FA. Foraminifera and nematodes can synthesize arachidonic acid (AA), eicosapentaenoic acid (EPA), while some barophylic bacteria produce EPA and/or docosahexaenoic acid. FA analysis of deep-sea invertebrates has shown high levels of PUFA including, in particular, arachidonic acid, bacterial FA, and a vast number of new and uncommon fatty acids such as 21:4(n-7), 22:4(n-8), 23:4(n-9), and 22:5(n-5) characteristic of foraminifera. We suppose that bacteria growing on detritus having a low trophic value provide the first trophic upgrading of organic matter for foraminifera and nematodes. In turn, these metazoans perform the second-stage upgrading for megafauna invertebrates. Deep-sea megafauna, including major members of Echinodermata, Mollusca, and Polychaeta display FA markers characteristic of bacteria, foraminifera, and nematodes and reveal new markers in the food chain.     

Interaction between Marine-Derived n-3 Long Chain Polyunsaturated Fatty Acids and Uric Acid on Glucose Metabolism and Risk of Type 2 Diabetes Mellitus: A Case-Control Study

The present case-control study explored the interaction between marine-derived n-3 long chain polyunsaturated fatty acids (n-3 LC PUFAs) and uric acid (UA) on glucose metabolism and risk of type 2 diabetes mellitus (T2DM). Two hundred and eleven healthy subjects in control group and 268 T2DM subjects in case group were included. Plasma phospholipid (PL) fatty acids and biochemical parameters were detected by standard methods. Plasma PL C22:6n-3 was significantly lower in case group than in control group, and was negatively correlated with fasting glucose (r = −0.177, p < 0.001). Higher plasma PL C22:6n-3 was associated with lower risk of T2DM, and the OR was 0.32 (95% confidence interval (CI), 0.12 to 0.80; p = 0.016) for per unit increase of C22:6n-3. UA was significantly lower in case group than in control group. UA was positively correlated with fasting glucose in healthy subjects, but this correlation became negative in T2DM subjects. A significant interaction was observed between C22:6n-3 and UA on fasting glucose (p for interaction = 0.005): the lowering effect of C22:6n-3 was only significant in subjects with a lower level of UA. In conclusion, C22:6n-3 interacts with UA to modulate glucose metabolism.     

水稻植株及其内生微生物磷脂脂肪酸生物标记异质性

采用改良磷脂脂肪酸法分析水稻植物自身磷脂脂肪酸和其内生微生物磷脂脂肪酸生物标记异质性。结果表明,水稻植株和其内生微生物共有磷脂脂肪酸有直链磷脂脂肪酸20∶0、12∶0、16∶0、18∶0和支链磷脂脂肪酸如18∶3ω6с(6,9,12)、19∶0 CYCLOω8с、15∶1ω6с等,且在水稻中含量均低于30μg/g,在内生微生物中的含量高于30μg/g。水稻内生微生物独有磷脂脂肪酸主要为支链磷脂脂肪酸,共有22种如i20∶0、20∶1ω9с、12∶0 3OH等,含量差异很大,为0.91～11 481.25μg/g。对水稻内生微生物独有磷脂脂肪酸对应分析CA得到,遗传距离近的水稻品种其对应距离也近,CA载荷图显示内生微生物磷脂脂肪酸对应距离小于姨2的为多元分布,即该磷脂脂肪酸分布在多个水稻品种中;大于姨2的为单一分布,即仅在特定的水稻品种中分布,表明水稻内生微生物磷脂脂肪酸分布与水稻品种遗传有关。     

Bioprospecting of Less-Polar Constituents from Endemic Brown Macroalga Fucus virsoides J. Agardh from the Adriatic Sea and Targeted Antioxidant Effects In Vitro and In Vivo (Zebrafish Model)

The endemic brown macroalga Fucus virsoides J. Agardh from the Adriatic Sea was in the focus of the present research. The volatiles of fresh (FrFv) and air-dried (DrFv) samples of F. virsoides obtained by headspace solid-phase microextraction (HS-SPME) and hydrodistillation (HD) were analyzed by gas chromatography equipped with flame ionization detector and mass spectrometry (GC-FID/MS). The major HS-FrFv compound was pentadecane (61.90–71.55%) followed by pentadec-1-ene (11.00–7.98%). In HS-DrFv, pentadec-1-ene was not present, and few lower aliphatic compounds appeared, as well as benzaldehyde and benzyl alcohol. In HD-FrFv, particularly abundant were alkenes (such as pentadec-1-ene (19.32%), or (E)-pentadec-7-ene (8.35%)). In HD-DrFv, more oxidation products were present (e.g., carbonyl compounds such as tridecanal (18.51%)). The fatty acids profile of freeze-dried sample (FdFv) after conversion to methyl esters was determined by GC-FID, and oleic acid was dominant (42.28%), followed by arachidonic acid (15.00%). High-performance liquid chromatography-high-resolution mass spectrometry with electrospray ionization (HPLC-ESI-HRMS) was used for the screening of less polar fractions (F3 and F4) of F. virsoides. Mono- and diglycerides of stearic, palmitic, oleic, and arachidonic acids were found. Terpenoids and steroids comprised the compounds C₂₀H₃₀₍₃₂₎O₂ and C₂₉H₄₈O₍₂₎. Among carotenoids, fucoxanthin was identified. Chlorophyll derivatives were also found (C₅₅H₇₄₍₇₂₎N₄O_(5-7)), dominated by pheophytin a. The antioxidant activity of the fractions was investigated by in vitro assays (oxygen radical absorbance capacity (ORAC), reduction of radical cation (ABTS•⁺), 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, and ferric reducing antioxidant power (FRAP)) and by in vivo zebrafish model (along with fish embryotoxicity). In vitro experiments proved good radical scavenging abilities of F3 and F4 fractions, which were additionally supported by the protective effect against hydrogen peroxide-induced oxidative stress in zebrafish embryos.     

GC-MS测定绿茶及速溶绿茶的游离脂肪酸

以绿茶及速溶绿茶为材料 ,采用硫酸_甲醇甲酯化 ,经GC_MS能够较好地分离和分析茶叶中的游离脂肪酸 ,三氟硼酸甲酯化不适合茶叶脂肪酸的分析。速溶绿茶中游离脂肪酸的含量十分少 ,只有绿茶原料的0 5 %。     

茶籽中脂肪酸的组成及其生物活性脂肪酸对高糖胁迫下RF/6A细胞生长的影响

采用气相色谱(GC)对茶籽的脂肪酸组成进行分析,并研究了活性脂肪酸对高浓度葡萄糖胁迫下RF/6A细胞生长的影响及其与葡萄糖对RF/6A细胞脂肪酸代谢的调节。结果表明,油酸与亚油酸为茶籽中含量最高的脂肪酸;而茶籽中脂肪酸的组成与其品种和产地有密切关系。亚油酸与亚麻酸对高浓度葡萄糖诱导的RF/6A细胞过度增殖有抑制作用。亚油酸、亚麻酸及棕榈酸对RF/6A细胞的脂肪酸代谢均具有调节作用。     

利用磷脂脂肪酸(PLFAs)生物标记法分析水稻根际土壤微生物多样性

采用磷脂脂肪酸(PLFAs)生物标记法,分析了4个以Ⅱ-32A为母本和3个以制5为父本的系列杂交水稻组合在同等栽培条件下根际土壤的微生物群落结构,并探讨其与水稻特性的内在联系,以期为水稻育种和品种改良提供科学依据。结果表明,水稻根际土壤磷脂脂肪酸生物标记丰富,共检测到38种PLFAs,其中完全分布的生物标记有23种,不完全分布的有15种;水稻根际土壤中PLFAs含量细菌真菌放线菌。不同水稻材料根际土壤微生物磷脂脂肪酸生物标记组成结构存在差异。对根际土壤特征磷脂脂肪酸生物标记16:0(细菌)、18:1ω9c(真菌)、10Me17:0(放线菌)、10Me16:0(硫酸盐还原细菌)和16:1ω5c(甲烷氧化菌)比较可知,制5系列组合含量高,最高的为金生优制5,Ⅱ-32A系列组合较低,含量最低的为Ⅱ优797。对PLFAs总量和水稻特性进行Pearson相关分析,结果表明,PLFAs含量与水稻有效穗数和产量呈显著正相关,但与株高呈显著负相关。对水稻根际土壤磷脂脂肪酸生物量总量进行聚类分析发现,Ⅱ-32A系列组合聚为一类,制5系列组合聚为一类,表现出水稻遗传特性在其根际PLFAs上的差异性。研究结果表明,水稻根际土壤微生物群落PLFAs结构丰富,根际土壤微生物群落结构受水稻遗传亲缘关系的影响。     

基于GC-MS法的离心浓缩椰浆风味成分及脂肪酸组成分析

选取气相色谱-质谱仪对离心浓缩椰浆样品的挥发性香气物质和脂肪酸的类型、含量进行研究。结果表明,共检测出20种香气成分,分为7大类化合物,包括酯类7种、有机酸3种、酮类3种、醇类4种、醛类1种、醚类1种、芳香族化合物1种。在检出的香气物质中,酯类所占比例最大,达60.50%,其次为酸类,所占比例为15.52%,酮类所占比例为12.78%。共鉴别出脂肪酸组分8种,其中饱和脂肪酸5种,约占总含量的72.31%,分别是葵酸(8.40%)、月桂酸(54.60%)、肉豆蔻酸(18.99%)、棕榈酸(8.68%)和硬脂     

Inventory of Fatty Acid Desaturases in the Pennate Diatom Phaeodactylum tricornutum

The diatom Phaeodactylum is rich in very long chain polyunsaturated fatty acids (PUFAs). Fatty acid (FA) synthesis, elongation, and desaturation have been studied in depth in plants including Arabidopsis, but for secondary endosymbionts the full picture remains unclear. FAs are synthesized up to a chain length of 18 carbons inside chloroplasts, where they can be incorporated into glycerolipids. They are also exported to the ER for phospho- and betaine lipid syntheses. Elongation of FAs up to 22 carbons occurs in the ER. PUFAs can be reimported into plastids to serve as precursors for glycerolipids. In both organelles, FA desaturases are present, introducing double bonds between carbon atoms and giving rise to a variety of molecular species. In addition to the four desaturases characterized in Phaeodactylum (FAD2, FAD6, PtD5, PtD6), we identified eight putative desaturase genes. Combining subcellular localization predictions and comparisons with desaturases from other organisms like Arabidopsis, we propose a scheme at the whole cell level, including features that are likely specific to secondary endosymbionts.