产油微藻自养培养条件调控

作为新兴生物燃料的生物柴油近年来发展迅速,以微藻为代表的第二代生物能源是解决能源危机的长远之计,但如何提高其产量仍是研究的热点问题。以提高产油自养微藻生物量和油脂含量为目的,在气升式光反应器中运用均匀设计实验方法进行了条件优化试验。分别得出了氮原子浓度、通气速率、二氧化碳体积浓度和光照强度4个因素对小球藻C2生物量积累和油脂含量影响的显著回归方程和反应器优化培养条件。以生物量为指标的优化培养条件是:氮原子浓度0.178 g/L,通气速率5 L/min,二氧化碳体积浓度3%(V/V),光照强度6000 lx。该优化条件下,生物量为2.11 g/L,即生产速率为0.352 g/(L.d),比测试实验中检测到的最高生物量[1.88 g/L,即生产速率为0.313 g/(L.d)]提高了12.2%;以油脂含量为指标的优化培养条件是:进气速率0.400 L/min,二氧化碳体积浓度1.94%(V/V),得到油脂含量为22.4%,比测试实验中检测到的最高油脂量(20.7%)提高7.7%。     

Fe~(3+)对小球藻的生长及油脂含量的影响

传统化石能源储量日益减少,生物柴油因其环保可再生性成为优质的石化柴油替代品。利用小球藻生产生物柴油速度快、油脂含量高,受到了广泛关注。为进一步提高小球藻生产生物柴油效率,分别探究了Fe3+的浓度及添加时间对自养和异养小球藻生长及产油的影响,获得最优Fe3+培养条件为:自养小球藻延滞期添加10-3 g/L Fe3+,生物量及油脂含量达2.80 g/L及30.90%;异养小球藻指数期添加10-5 g/L Fe3+,生物量及油脂含量达3.30 g/L及29.05%。经脂肪酸分析,以上条件获得的微藻油脂均可作为生物柴油生产原料。     

小球藻与固氮菌Mesorhizobium sp.共培养对小球藻生长和油脂积累的促进效果

微藻油脂不仅可以作为功能油脂,同时也是生产生物柴油的重要原料之一。为解决微藻生长与油脂积累之间的矛盾,利用藻菌共培养技术在缺氮条件下将无菌小球藻与细菌以不同初始比例进行共培养,通过测定藻细胞生物量、油脂含量和脂肪酸比例等来研究藻菌共培养对小球藻生长和油脂积累的影响。结果表明,在小球藻与固氮菌B2. 3 70∶1(V/V)共培养体系中,小球藻的生物量和油脂含量较同样条件下单独培养小球藻有了显著提高。其生物量最高可达1. 68g/L、总脂含量为45. 2%、总脂产率为75. 94 mg/(L·d)、中性脂含量为23. 0%及中性脂产率为38. 65mg/(L·d),其生物量和油脂含量分别较单独小球藻培养时提高了66. 3%和47. 7%。同时细菌的加入显著提高了藻细胞内C18∶1脂肪酸的比例。结论表明,通过藻菌共培养技术能够有效提高微藻生物油脂的质量和产量,具有较好的实际利用价值。     

氮胁迫对埃氏小球藻生长及油脂积累的影响

以生长快、可除污的埃氏小球藻株系SXND-25为试材,研究不同氮浓度培养条件对其生物量和油脂产量的影响,以期建立优化培养体系利用该株小球藻生产优质生物燃油。以硝酸钠为氮源、BG11培养基中的氮浓度为基准(1.5 g/L),设置氮浓度梯度对小球藻进行培养。通过光密度测定、尼罗红染色、转酯化法抽提油脂和GC分析,对小球藻生物量、油脂含量及脂肪酸组分进行分析。结果显示,培养8 d时,在氮浓度为1.5 g/L时,生物量达到最大,干重为3.4 g/L,而油脂含量仅为28.24%,油脂产量为0.96 g/L;在氮浓度为0 g/L时生物量最小,干重为0.49 g/L,而油脂含量最高,为44.57%,油脂产量为0.22 g/L;在氮浓度为0.75 g/L时,干重为3.2 g/L,油脂含量为40.36%,油脂产量最高为1.3 g/L,是标准氮浓度下油脂产量的1.4倍。0.75 g/L氮浓度下连续培养8 d,藻油脂肪酸组成更适于制取优质生物柴油。综合生物量、油脂含量及脂肪酸组成等指标,确定0.75 g/L氮浓度为该埃氏小球藻株系规模化培养以生产优质生物燃油的优化参数。     

氮源对C.vulgaris油脂积累的影响

实验室条件下,考察了在发酵过程中不同氮源对小球藻的生物量和油脂积累的影响,确定了小球藻的最佳氮源;并对比分析了含氮培养与缺氮培养的生物量、油脂含量、氮消耗量、生物量氮消耗比率和油脂氮消耗比率的不同。结果表明:小球藻在1.6 g/L Na NO3时获得最大生物量,为562.2 mg/L,在0.8 g/L Na NO3时获得最大相对油脂含量为12.01%;以油脂含量为考察指标时,培养小球藻的最佳氮源为0.8 g/L Na NO3;缺氮培养时,最大油脂含量为13.49%,比含氮培养高约15%;含氮培养时,最高生物量为626.3 mg/L,比缺氮培养高约1.9倍。氮源对生物量,相对油脂含量,生物量氮消耗比率和油脂氮消耗比率具有明显的影响。藉此,提出了通过改变培养方式,达到调控小球藻细胞内生理代谢组分的可行性。     

蛋白核小球藻发酵产油脂的研究

从5种不同来源的小球藻中筛选到1株油脂产量较高的蛋白核小球藻Chlorella pyrenoidosa No.2。研究了培养基组成及培养条件对其细胞生长和油脂积累的影响。结果表明, 最适培养基组成为(g/L)：葡萄糖 20, 甘氨酸 0.08, MgSO4·7H2O 0.4, K2HPO4 1.0, FeSO4·7H2O 0.004; 适宜的培养温度、初始pH、摇床转速和光照强度分别为28℃、6.0、130 r/min和 650 Lux。在上述优化条件下培养7 d, Chlorella pyrenoidosa No.2的生物量和油脂含量分别由优化前的3.73 g/L 和 40.15%提高到6.56 g/L和59.90%, 油脂产量提高了162%。Chlorella pyrenoidosa No.2能以木糖为碳源产油脂, 可望用于以木质纤维素等可再生生物质资源为原料生产油脂。气相色谱分析表明该油脂的脂肪酸组成与植物油相似, 不饱和脂肪酸含量达71％左右, 可作为生产生物柴油的原料。     

不同培养基及组成对2种小球藻生长和油脂的影响

研究了4种培养基及组成对蛋白核小球藻F-9和普通小球藻HYS-2的生长、油脂积累和脂肪酸组成的影响。结果发现knop、Provasoli、f/2、MAV 4种培养基中,f/2培养基更有利于小球藻的快速生长,而MAV培养基更适合油脂积累。在f/2培养基中F-9和HYS-2相对生长速率分别为0.156和0.171,培养9 d细胞干重为0.188 g/L和0.195 g/L。而在MAV培养基中F-9油脂含量最高可达19.67%,HYS-2油脂含量最高为21.91%,脂肪酸最高分别占干重的5.11%和8.71%。N/P为16∶1时小球藻生长最快,培养9 d后F-9和HYS-2的相对生长速率分别为0.23和0.239,最终细胞干重分别为0.107 g/L和0.143 g/L。而F-9和HYS-2在N/P为1∶1条件下积累油脂和脂肪酸含量最高,总脂含量分别占干重的为20.40%和27.39%,总脂肪酸占藻粉干重的含量为12.52%和16.94%。     

蛋白核小球藻发酵产油脂的研究

从5种不同来源的小球藻中筛选到1株油脂产量较高的蛋白核小球藻Chlorella pyrenoi-dosa No.2.研究了培养基组成及培养条件对其细胞生长和油脂积累的影响.结果表明,最适培养基组成为(g/L):葡萄糖20,甘氨酸0.08,MgSO4·7H2O 0.4,K2HPO4 1.0,FeSO4·7H2O 0.004;适宜的培养温度,初始pH、摇床转速和光照强度分别为28℃、6.0、130 r/min和650 Lux.在上述优化条件下培养7 d,Chlorella pyrenoidosa No.2的生物量和油脂含量分别由优化前的3.73 g/L和40.15%提高到6.56 g/L和59.90%,油脂产量提高了162%.Chlorella pyrenoidosa No.2能以木糖为碳源产油脂,可望用于以木质纤维素等可再生生物质资源为原料生产油脂.气相色谱分析表明该油脂的脂肪酸组成与植物油相似,不饱和脂肪酸含量达71%左右,可作为生产生物柴油的原料.     

氧化铝气体分布器应用小球藻培养的研究

光生物反应器设计中,气体分布器对微藻生长有较大的影响,尤其是在鼓泡式光生物反应器中更为显著。实验考察了采用氧化铝烧制的多孔气体分布器的5L鼓泡式光生物反应器中通气速率、CO₂ 浓度对小球藻LICME002生物量、叶绿素含量、油脂积累的影响。对该气体分布器下的CO₂浓度和通气速率对小球藻的作用机理进行了初步的探讨。结果表明,CO₂浓度为3%时,该株微藻生物量、叶绿素、油脂积累的最佳;CO₂浓度超过6%时各项指标显著下降。通过对0.1vvm,0.4vvm,0.7vvm、1.0vvm的通气速条件下的各项指标的分析,确定最佳通气条件为0.4vvm。结论显示,在最佳通气速率和CO₂浓度下,微藻生物量能达到1.52g/L,油脂含量达到31.5%。     

不同氮源对异养小球藻生物量和油脂积累的影响

小球藻因其快速生长和易培养等特性可用于制备生物能源。与传统的光自养相比,异养小球藻可获得更多的生物量和更高的油脂含量。低成本的马铃薯淀粉水解液可作为小球藻的理想碳源,在氮饥饿条件下可诱导产生更多的油脂。为了探讨不同氮源对异养小球藻生物量和油脂积累的影响,并筛选出异养条件下的最适氮源,实验研究了不同浓度无机氮源NaNO3以及有机氮源丙氨酸和酪氨酸对异养小球藻生物量和油脂积累的影响。以马铃薯淀粉水解液为唯一碳源,在SE培养基中分别添加不同氮源培养小球藻。设定的NaNO3和丙氨酸浓度均为1.5 mmol/L、3.0 mmol/L、6.0 mmol/L,酪氨酸浓度为0.75 mmol/L、1.5 mmol/L和3.0mmol/L。所有小球藻培养实验均为暗培养并持续10 d时间。实验过程测定的指标为:小球藻的细胞数目、比生长速率、叶绿素含量、中性脂含量和总脂含量。实验结果表明:(1)在异养条件下以硝酸盐为无机氮源时,氮源促进叶绿素积累从而促进小球藻的生长,减少硝态氮可以使小球藻快速进入稳定期积累油脂。在NaNO3中氮含量为1.5 mmol/L时,生物量和油脂含量分别为2.65 g/L和51.21%,总油脂含量为1.36 g/L。(2)在不添加其他氮源的异养培养基中,丙氨酸可促进小球藻的生物量增加,在稳定期仍促进单位细胞的叶绿素含量,但总油脂含量普遍偏低。(3)酪氨酸可抑制小球藻生物量增加,使细胞膨大从而促进单位细胞内叶绿素和油脂合成,油脂含量高达38.78%—47.02%。这些结果表明小球藻可通过诱导氨基酸转运系统适应氮源的变化,其中酪氨酸所在的第三个转运系统在葡萄糖诱导条件下可促进油脂的合成。     

Polymorphism of 1-O-alkylglycerols and general lipid polymorphic behaviour

The polymorphic behaviour of a series of l-O-alkylglycerols have been studied; mainly by DTA and X-ray powder diffraction techniques. The behaviour of these ether lipids is quite similar to that of the closely related l-acylglycerols. The background to the polymorphic behaviour of these lipids is discussed. In the α-phase, the alkyl chains are non-crystalline but still partly ordered in a lattice. The main reason for the formation of the sub-α-phase is a ‘chain crystallization’. The polar group region in the sub-α-phase is crystallized in a state with higher energy, relative to the state of the polar group region in the β-phase. This can account for the difference in stability the β - and sub-ga-phases.     

Functions and interaction of plant lipid signalling under abiotic stresses

Lipids are the primary form of energy storage and a major component of plasma membranes, which form the interface between the cell and the extracellular environment. Several lipids — including phosphoinositide, phosphatidic acid, sphingolipids, lysophospholipids, oxylipins, and free fatty acids — also serve as substrates for the generation of signalling molecules. Abiotic stresses, such as drought and temperature stress, are known to affect plant growth. In addition, abiotic stresses can activate certain lipid-dependent signalling pathways that control the expression of stress-responsive genes and contribute to plant stress adaptation. Many studies have focused either on the enzymatic production and metabolism of lipids, or on the mechanisms of abiotic stress response. However, there is little information regarding the roles of plant lipids in plant responses to abiotic stress. In this review, we describe the metabolism of plant lipids and discuss their involvement in plant responses to abiotic stress. As such, this review provides crucial background for further research on the interactions between plant lipids and abiotic stress.     

Caveolin-1-dependent and -independent membrane domains

Lipid rafts defined as cholesterol- and sphingomyelin-rich domains have been isolated from different cell types that vary greatly in their lipid profiles. Here, we investigated the contribution of the structural protein caveolin-1 (Cav1) to the overall lipid composition and domain abundance in mouse embryonic fibroblasts (MEFs) from wild-type (WT) or Cav1-deficient (Cav1^−/−) animals. Our findings show that Cav1 expression had no effect on free (membrane-associated) cholesterol levels. However, Cav1^−/−-deficient cells did have a higher proportion of sphingomyelin, decreased abundance of unsaturated phospholipids, and a trend toward shorter fatty acid chains in phosphatidylcholine. We isolated detergent-resistant membranes (DRMs), nondetergent raft domains (NDR), and cholesterol oxidase (CO)-sensitive domains and assessed the abundance of ordered domains in intact cells using the fluorescent dye Laurdan. Despite differences in phospholipid composition, we found that cholesterol levels in DRMs, NDR, and CO-sensitive domains were similar in both cell types. The data suggest that Cav1 is not required to target cholesterol to lipid rafts and that CO does not specifically oxidize caveolar cholesterol. In contrast, the abundance of ordered domains in adherent cells is reduced in Cav1^−/− compared with WT MEFs, suggesting that cell architecture is critical in maintaining Cav1-induced lipid rafts.     

The influence of proteins and peptides on the phase properties of lipids

This paper reviews model membrane studies on the modulation of the macroscopic structure of lipids by lipid-protein interactions, with particular emphasis on the gramicidin molecule. This hydrophobic peptide has three main effects on lipid polymorphism: (1) in lysophosphatidylcholine it triggers a micellar to bilayer transition, (2) in phosphatidylethanolamine it lowers the bilayer to hexagonal H_II phase transition temperature and (3) in phosphatidylcholine and other bilayer preferring lipids it is able to induce the formation of an H_II phase. From experiments in which the gramicidin molecule was chemically modified it can be concluded that the tryptophan residues play a determining role in the peptide-induced changes in polymorphism. The experimental data lead to the proposal that gramicidin molecules have a tendency to self-associate, possibly mediated by tryptophan-tryptophan interactions and organize into tubular structures such as found in the H_II phase.     

Import and fate of fluorescent analogs of oxidized phospholipids in vascular smooth muscle cells

Lipid oxidation is now thought to be an initiating and sustaining event in atherogenesis. Oxidatively fragmented phospholipids, namely 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC) and 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC), present in minimally modified LDL and atherosclerotic lesions, have been reported to elicit a wide range of pathophysiological responses in the cells of the vascular wall. Nevertheless, the question of their potential sites of action and their primary molecular targets remains open. To address this issue, a series of fluorescently labeled analogs, which differ with regard to structure and binding site of the fluorophore, were synthesized and used as tools for studying the uptake, intracellular stability, and distribution of PGPC and POVPC in vascular smooth muscle cells (VSMCs). We demonstrate that in accordance with their lysophospholipid-like structure, these highly similar molecules transferred rapidly either from aqueous phospholipid dispersions or preloaded native LDL into VSMCs, producing disparate fluorescence patterns irrespective of the attached fluorophore. PGPC derivatives were translocated to the lysosomes. In sharp contrast, POVPC analogs were initially captured in the plasma membrane, most likely in consequence of the formation of covalent adducts with free amino and sulfhydryl groups of proteins and phospholipids. LDL internalization is not required for cellular lipid uptake. Collectively, our data provide evidence that oxidized phospholipids, owing to their high exchangeability between lipoproteins and cell membranes, may act within a short time on different cellular sites in VSMCs and affect various lipid and protein components through physical or chemical interactions, which might then serve as starting points for intracellular signaling.     

Lipid polymorphism and the roles of lipids in membranes

The reasons for lipid diversity in membranes are not understood. Here we review evidence supporting the proposal that factors related to the polymorphic capabilities of lipids provide a rationale for lipid diversity. In particular, the ability of lipids to adopt different polymorphic phases appears to be related to a generalized shape property, where lipids with a cylindrical geometry preferentially adopt the bilayer phase whereas ‘cone’ shaped lipids adopt the hexagonal H_II phase. Lipid diversity may then be considered to satisfy three demands. The first is obviously a need for bilayer forming lipids to provide the basic permeability barrier, whereas the second concerns a need for non-bilayer lipids and associated structures for fusion and related membrane contact phenomena to proceed. A third, and less obvious demand satisfied by non-bilayer lipids concerns the ability of lipids of different shapes to modulate the order in the hydrocarbon region when constrained to a bilayer organization. These possibilities are summarized in a metamorphic mosaic model of membranes.     

Three-dimensional enhanced lipidomics analysis combining UPLC,differential ion mobility spectrometry,and mass spectrometric separation strategies

Phospholipids serve as central structural components in cellular membranes and as potent mediators in numerous signaling pathways. There are six main classes of naturally occurring phospholipids distinguished by their distinct polar head groups that contain many unique molecular species with distinct fatty acid composition. Phospholipid molecular species are often expressed as isobaric species that are denoted by the phospholipid class and the total number of carbon atoms and double bonds contained in the esterified fatty acyl groups (e.g., phosphatidylcholine 34:2). Techniques to separate these molecules exist, and each has positive and negative attributes. Hydrophilic interaction liquid chromatography uses polar bonded silica to separate lipids by polar head group but not by specific molecular species. Reversed phase (RP) chromatography can separate by fatty acyl chain composition but not by polar head group. Herein we describe a new strategy called differential ion mobility spectrometry (DMS), which separates phospholipid classes by their polar head group. Combining DMS with current LC methods enhances phospholipid separation by increasing resolution, specificity, and signal-to-noise ratio. Additional application of specialized information-dependent acquisition methodologies along with RP chromatography allows full isobaric resolution, identification, and compositional characterization of specific phospholipids at the molecular level.     

A non-destructive morphometric technique for estimation of body and mesenteric lipid in Arctic charr: a case study of its application

Weight and eight linear measurements were made on Arctic charr from the domesticated Hammerfest strain from Norway and offspring of wild charr of a pelagic morph from Loch Rannoch, Scotland. Guts and mesenteries were removed from the Hammerfest charr only, and the amount of lipid in both carcass and mesenteries measured by Soxhlet extraction. Lipid was extracted from the whole body of the Loch Rannoch charr. Multiple regression analysis was used to derive morphometric predictors of total lipid for the Hammerfest charr and percentage body lipid for the Loch Rannoch charr, the regressions explaining 83 and 59% of variance respectively. For the Hammerfest charr, multiple regression also provided a reliable predictor of mesenteric fat, accounting for 65% of its variance. Hammerfest charr that exhibited high aggression rates had 53% more whole body lipid and 100% more mesenteric fat than those with low aggression rates, using direct measures of lipid levels. Indirect, morphometrically-derived measures of lipid levels gave almost identical results. It is concluded that morphometric techniques can provide estimates of both whole body and mesenteric lipid in studies requiring repeated measures on the same individuals.