乙酰水杨酸对三角褐指藻岩藻黄质含量的影响及其分子机理研究

为了研究不同浓度乙酰水杨酸(ASA)对三角褐指藻(Phaeodactylum tricornutum Bohlin)细胞生长、岩藻黄质积累及岩藻黄质生物合成相关基因的表达水平的影响,通过分光光度计、高效液相色谱(HPLC)以及荧光定量PCR测定不同浓度ASA处理后的三角褐指藻的各项指标。结果表明,三角褐指藻的生长和岩藻黄质积累与ASA浓度密切相关。一定浓度的ASA抑制了三角褐指藻的细胞数量;岩藻黄质含量也随着ASA浓度的升高呈现先上升后下降的趋势,当ASA浓度为10 mg·L^-1时,岩藻黄质含量最高,达1.78 g·kg^-1 DW,为对照组的1.9倍。在不同浓度ASA诱导下,与岩藻黄质生物合成途径有关的PSY、PDS、ZDS、CRTISO、LCYB和ZEP基因表达均上调,当ASA浓度为10 mg·L^-1时,相关基因的表达量极显著高于对照组(P<0.01)。主成分分析(PCA)结果表明,PSY、PDS、ZDS、CRTISO、LCYB和ZEP基因与岩藻黄质积累具有相关性,其中,ZEP基因对岩藻黄质生物合成贡献率最高。综...     

三角褐指藻作为饲料蛋白质的开发前景研究

三角褐指藻是一种单细胞蛋白,蛋白质含量丰富,质量好,动物易消化,并且三角褐指藻容易培养,生长速度快。本文旨在通过测定藻液中的主要营养指标蛋白质的变化,探讨三角褐指藻藻液的采收,研究三角褐指藻作为蛋白质饲料添加到饲料中的可能性,为饲料蛋白质提供新的来源。     

Heterotrophic Production of Omega-3 Long-Chain Polyunsaturated Fatty Acids by Trophically Converted Marine Diatom Phaeodactylum tricornutum

We have created via metabolic engineering a heterotrophic strain of Phaeodactylum tricornutum that accumulates enhanced levels of the high value omega-3 long chain polyunsaturated fatty acid (LC-PUFAs) docosahexaenoic acid (DHA). This was achieved by generation of transgenic strains in which the Δ5-elongase from Ostreococcus tauri was co-expressed with a glucose transporter from the moss Physcomitrella patens. This double transformant has the capacity to grow in the dark in liquid medium supplemented with glucose and accumulate substantial levels of omega-3 LC-PUFAs. The effects of glucose concentrations on growth and LC-PUFA production of wild type and transformed strains cultivated in the light and dark were studied. The highest omega-3 LC-PUFAs accumulation was observed in cultures grown under mixotrophic conditions in the presence of 1% glucose (up to 32.2% of total fatty acids, TFA). Both DHA and EPA are detected at high levels in the neutral lipids of transgenic cells grown under phototrophic conditions, averaging 36.5% and 23.6% of TFA, respectively. This study demonstrates the potential for P. tricornutum to be developed as a viable commercial strain for both EPA and DHA production under mixo- and heterotrophic conditions.     

Marine Benthic Diatoms Contain Compounds Able to Induce Leukemia Cell Death and Modulate Blood Platelet Activity

In spite of the high abundance and species diversity of diatoms, only a few bioactive compounds from them have been described. The present study reveals a high number of mammalian cell death inducing substances in biofilm-associated diatoms sampled from the intertidal zone. Extracts from the genera Melosira, Amphora, Phaeodactylum and Nitzschia were all found to induce leukemia cell death, with either classical apoptotic or autophagic features. Several extracts also contained inhibitors of thrombin-induced blood platelet activation. Some of this activity was caused by a high content of adenosine in the diatoms, ranging from 0.07 to 0.31 μg/mg dry weight. However, most of the bioactivity was adenosine deaminase-resistant. An adenosine deaminase-resistant active fraction from one of the extracts was partially purified and shown to induce apoptosis with a distinct phenotype. The results show that benthic diatoms typically found in the intertidal zone may represent a richer source of interesting bioactive compounds than hitherto recognized.     

Specific Metabolites in a Phaeodactylum tricornutum Strain Isolated from Western Norwegian Fjord Water

We have searched for special characteristics in growth, protein expression, fatty acids and volatile organic compounds (VOCs) in a local Phaeodactylum tricornutum Bohlin strain (Bergen Marine Biobank), by comparing it with a common accession strain (CCAP). Differences in growth and expressed proteins were detected between the BMB strain and the CCAP strain, and the BMB strain reached the highest cell densities under the given growth conditions. Fatty acid (FA) analyses showed highest relative eicosapentaenoic acid (EPA) levels in the exponential phase (25.73% and 28.31%), and highest levels of palmitoleic acid (16:1 n-7) in the stationary phase (46.36% and 43.66%) in the BMB and CCAP strain, respectively. The most striking finding of the VOCs analyses was the relatively high levels of ectocarpene, 6-((1E)-butenyl)-1,4-cycloheptadiene, hormosirene, and desmarestene and structurally related compounds, which were exclusively detected in the BMB strain. Many of the VOCs detected in the CCAP and, in particular, in the BMB strain have been reported as antimicrobial agents. We suggest that the array of pheromones and antimicrobial substances could be part of an allelopathic strategy of the BMB strain, dominated by oval cells, thus reflecting the benthic life stage of this morphological form. These findings show the potential for bioactive metabolites in the BMB strain.     

三角褐指藻细胞破碎方法的研究

三角褐指藻是一种含有较高多不饱和脂肪酸的海洋单胞藻。为了从细胞中提取不饱和脂肪酸,采用反复冻融法、超声波破碎法和匀浆法分别对三角褐指藻细胞进行破碎,并测定油脂提取率。结果表明,超声波破碎法、反复冻融法和匀浆法的最大细胞破碎率分别为91.5%,56.7%和81.2%;油脂提取率分别为18.1%,13%和13.4%。从操作的简捷性和经济效益来看,超声波破碎法优于反复冻融法和匀浆法。     

Rapid Sorting of Fucoxanthin-Producing Phaeodactylum tricornutum Mutants by Flow Cytometry

Fucoxanthin, which is widely found in seaweeds and diatoms, has many benefits to human health, such as anti-diabetes, anti-obesity, and anti-inflammatory physiological activities. However, the low content of fucoxanthin in brown algae and diatoms limits the commercialization of this product. In this study, we introduced an excitation light at 488 nm to analyze the emitted fluorescence of Phaeodactylum tricornutum, a diatom model organism rich in fucoxanthin. We observed a unique spectrum peak at 710 nm and found a linear correlation between fucoxanthin content and the mean fluorescence intensity. We subsequently used flow cytometry to screen high-fucoxanthin-content mutants created by heavy ion irradiation. After 20 days of cultivation, the fucoxanthin content of sorted cells was 25.5% higher than in the wild type. This method provides an efficient, rapid, and high-throughput approach to screen fucoxanthin-overproducing mutants.     

3种不同来源的三角褐指藻的生长与油脂积累差异

[目的]研究3种不同来源的三角褐指藻的生长与油脂积累差异,为藻种的选择和培养工艺的确定提供参考。[方法]比较不同来源的三角褐指藻QEAC501、QEAC503和QEAC504在实验室条件下的生长与油脂积累能力,以及主要的脂肪酸的碳链组成,[结果]3个三角褐指藻品种在正常的摇瓶批次培养条件下生长速度无明显差异,同时由于营养供应的充足,都不能大量的积累脂类物质;而经过营养缺乏的诱导,3个品种均明显出现大量的脂质尤其是甘油三脂的积累,其中品种间甘油三脂的积累能力有明显差异,脂肪酸链长组成总体上没有明显差异,均以C-16为主。[结论]利用薇藻生产油脂时应注意选择高油脂积累的品种,并在生产过程中设置营养缺乏诱导以提高薇藻的油脂积累。