紫外诱变对微绿球藻生长和营养成分的影响

微绿球藻是一种海洋单细胞微藻,含有丰富的EPA、DHA等活性代谢产物,为了获得生长快、高产EPA的优良藻株,利用紫外诱变技术对微绿球藻进行诱变,筛选出生长较快的突变株2株（MN-1和MN-2）,并与出发株就生物量、粗蛋白、多糖、总脂及脂肪酸进行了比较。结果表明：突变株MN-1与出发株相比,生长速率增加21．08％,生物量增加14．55％,粗蛋白含量增加2．54％,总脂含量增加9．81％,EPA增加1．81％,SFA增加2．70％;突变株MN-2的生长率增加6．25％,生物量增加5．62％,多糖增加13．26％,总脂增加7．93％,SFA增加28．9％。     

高产油小球藻的筛选及其油脂分析

小球藻广泛分布于各种生境,特别是淡水环境中,适应性强。其同化产物主要是淀粉,但在环境胁迫条件下可显著积累中性脂,其脂肪酸类型主要为C16和C18,适合作为生物柴油的原料。我们从中国部分地区水体中分离纯化到若干株小球藻,通过薄层层析比较分析了21株产油小球藻的油脂含量,筛选到一株三酰基甘油含量较高的藻株Chlorella sp.NMX37N。其适宜生长温区为15—35℃,在25℃时生长速率最快,比生长速率为0.53/d,生长的最适光强为250μmol photons/(m2.s)。批量培养实验显示,藻细胞的三酰基甘油含量随培养时间延长而增加,并在培养的稳定期达到最大值,此时培养液中氮基本被耗尽。在批量培养条件下培养Chlorella sp.NMX37N约40d,藻细胞中总脂含量可达到33%左右,与此相比通过两步培养方式,将培养至对数后期(约20d)的藻细胞缺氮处理48h后,得到的总脂产率相当。通过两步培养方式可以大大缩短培养时间,使得该藻细胞快速有效积累油脂。另外,气相色谱分析显示,该藻的总脂和三酰基甘油的脂肪酸均以C16∶0和C18∶2为主,占总脂肪酸的70%以上,且不含C20以上的长链脂肪酸,可以作为优质的生物柴油原料。     

微拟球藻属对盐度的耐受及其产油特性分析

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不同氮源及CO_2浓度下湖泊微拟球藻的生长及产油特性分析

<正>工业生产排放的各种废气与污水污染人类赖以生存的环境。化石燃料燃烧除排放出大量CO2外,还释放出含有粉尘、SO[x和NOx等直接危害人类健康的有毒成分1]。其中NO]x与水结合后最终会转化成硝酸盐或亚硝酸盐等[2,导致污水中的含氮化合物氨氮、亚硝酸盐氮、硝酸盐氮和有机氮的含量通常偏高。如何有效去除污水中的氮是防治水体污染最关键的步骤之一,而利用微藻培养去除水体中的氮源是目前研究的热点。尽管生物燃料生产的成本远     

眼点拟微绿球藻生长的生态因子分析

眼点拟微绿球藻(Ｎａｎｎｏｃｈｌｏｒｏｐｓｉｓｏｃｕｌａｔａ)是重要的海水经济藻类,本文研究了温度,光强,盐度,碳因子对藻体生长的影响,并采用正交实验的方法研究了Ｎ,Ｐ,Ｆｅ三因子对其生长的影响。试验表明:眼点拟微绿球藻的生长适温范围０－３０℃,最适温度为２０℃;适光范围较广,喜强光,最适光强５ ０００－７ ０００ ｌｘ;在盐度为１３．７－３３．４范围生长率差别不明显,最适盐度为２６．９;碳源偏喜重碳酸盐。Ｎ浓度为１０μｍｏｌ·Ｌ－１,Ｐ浓度为２μｍｏｌ．Ｌ－１,Ｆｅ浓度为０．２μｍｏｌ·Ｌ－１时对该藻生长最有利。     

半导体激光辐照对拟微绿球藻的生物学效应

目的:研究半导体激光对拟微绿球藻的生物学效应,进而利用激光诱变筛选高产优质藻株.方法:采用半导体激光辐照拟微绿球藻,辐照时间为l 0min、20min、30min.结果:诱变后藻的代谢物产量均有提高.结论:l0min、20min、30min辐照条件下,对拟微绿球藻细胞生长及代谢产物均有不同程度的促进作用,其中10min辐照组有利于拟微绿球藻的生长及色素的积累,分别比对照组提高了45.06％、86.59％、56.06％;但在20min剂量下更有利于胞外多糖、总多糖、蛋白质及油脂含量的积累,分别比对照组提高了67.7％、51.89％、19.16％、69.78％.     

CO_2激光辐照拟微绿球藻的当代生物学效应研究

采用CO_2激光(波长10.6μm,功率10 w,光束长74 cm)辐照拟微绿球藻(Nannochloropsis sp.YW0980),辐照时间为30 s、60 s、90 s,通过测定藻色素、多糖、蛋白质及油脂含量,研究CO_2激光对藻的生物学效应。结果表明:30 s、60 s、90 s辐照条件下,对拟微绿球藻细胞生长及代谢产物均有一定的促进作用,其中CO_2激光60 s处理组有利于拟微绿球藻的生长及色素的积累,但30 s剂量下更有利于多糖、蛋白质及油脂含量的积累,分别比对照组提高了51.05%(胞外多糖),289.45%(总多糖)、37.05%、172.16%。     

CO2激光辐照拟微绿球藻的当代生物学效应研究

采用CO₂激光（波长10.6μm,功率10 w,光束长74 cm）辐照拟微绿球藻（Nannochloropsis sp.YW0980）,辐照时间为30 s、60 s、90 s,通过测定藻色素、多糖、蛋白质及油脂含量,研究CO₂激光对藻的生物学效应。结果表明:30 s、60 s、90 s辐照条件下,对拟微绿球藻细胞生长及代谢产物均有一定的促进作用,其中CO₂激光60 s处理组有利于拟微绿球藻的生长及色素的积累,但30 s剂量下更有利于多糖、蛋白质及油脂含量的积累,分别比对照组提高了51.05%（胞外多糖）,289.45%（总多糖）、37.05%、172.16%。     

眼点拟微绿球藻对抗生素的敏感性及其无菌藻株的培养

研究了5种抗生素在不同浓度下对眼点拟微绿球藻(Nannochloropsis ocutala)生长的影响。结果表明:浓度为50μg·mL^-1时,氯霉素对眼点拟微绿球藻生长的抑制作用最明显,抑制率高达85.30%,2d和4d的Ic₅₀分别为293.64μg·mL^-1和67.69μg·mL^-1。其次为青霉素、硫酸庆大霉素和硫酸卡那霉素,4d的抑制率分别为8.95%、7.62%和3.84%,其中硫酸庆大霉素2d的Ic₅₀为316.40μg·mL^-1。链霉素对藻细胞生长的抑制作用最轻,抑制率仅为3%。按照抗生素对眼点拟微绿球藻生长抑制作用,由强到弱依次加入50μg·mL^-1青霉素,50μg·mL^-1硫酸庆大霉素,100μg·mL^-1硫酸卡那霉素和50μg·mL^-1链霉素,获得了无菌藻株。     

湖泊微拟球藻高含油突变株的构建及筛选

研究旨在建立湖泊微拟球藻(Nannochloropsis)的遗传转化体系, 然后根据外源基因随机插入的特性, 挑取抗性转化子构建突变体库, 并从突变体库中筛选高含油突变株。以湖泊微拟球藻自身β-tublin基因启动子和三角褐指藻fcpA终止子驱动和终止来源于细菌的sh ble抗性选择基因, 构建了一个转化载体pPha-T1-TUB。将线性化后的质粒以电穿孔的方法转化湖泊微拟球藻, 通过1 μg/mL zeocin的抗性平板筛选, 并经液体培养基连续传代后, 得到了可以稳定遗传的转化子。我们从这些转化子中筛选得到了数株油含量高于野生型, 且生长也优于野生型的突变株。其中, 2个突变株K26和G5的总脂中多不饱和脂肪酸含量更低, 其脂肪酸组成更符合作为生物柴油原料的标准。研究通过随机插入构建突变体库的方法为快速获取优良目的性状的高产油突变株提供了一个有效手段。     

RADIOLABELING STUDIES OF LIPIDS AND FATTY ACIDS IN NANNOCHLOROPSIS (EUSTIGMATOPHYCEAE), AN OLEAGINOUS MARINE ALGA1

The synthesis of fatty acids and lipids in Nannochloropsis sp. was investigated by labeling cells in vivo with [¹⁴C]-bicarbonate or [¹⁴C]-acetate. [¹⁴C]-bicarbonate was incorporated to the greatest extent into 16:0, 16:1, and 14:0 fatty acids, which are the predominant fatty acids of triacylglycerols. However, more than half of the [¹⁴C]-acetate was incorporated into longer and more desaturated fatty acids, which are constituents of membrane lipids. [¹⁴C]-acetate was incorporated most strongly into phosphatidylcholine, which rapidly lost label during a 5-h chase period. The label associated with phosphatidylethanolamine also decreased during the chase period, whereas label in other membrane lipids and triacylglycerol increased. The dynamics of labeling, along with information regarding the acyl compositions of various lipids, suggests that 1) the primary products of chloroplast fatty acid synthesis are 14:0, 16:0, and 16:1; 2) C₂₀ fatty acids are formed by an elongation reaction that can utilize externally supplied acetate; 3) phosphatidylcholine is a site for desaturation of C₁₈ fatty acids; and 4) phosphatidylethanolamine may be a site for desaturation of C₂₀ fatty acids.     

FATTY ACIDS AND HYDROXY FATTY ACIDS IN THREE SPECIES OF FRESHWATER EUSTIGMATOPHYTES

The monocarboxylic fatty acids and hydroxy fatty acids of three species of freshwater microalgae—Vischeria punctata Vischer, Vischeria helvetica (Vischer et Pascher) Taylor, and Eustigmatos vischeri (Hulbert) Taylor, all from the class Eustigmatophyceae— were examined. Each species displayed a very similar distribution of fatty acids, the most abundant of which were 20:5n-3, 16:0, and 16:1n-7; C₁₈ polyunsaturated fatty acids were minor components. These fatty acid distributions closely resemble those found in marine eustigmatophytes but are quite distinct from those found in most other algal classes. These microalgae also contain long-chain saturated and unsaturated monohydroxy fatty acids. Two distinct types of hydroxy fatty acids were found: a series of saturated α-hydroxy acids ranging from C₂₄ to C₃₀ with a shorter series of monounsaturated α-hydroxy acids ranging from C₂₆ to C₃₀ together with a series of saturated β-hydroxy acids ranging from C₂₆ to C₃₀. The latter have not previously been reported in either marine or freshwater microalgae, although C₃₀ to C₃₄ midchain (ω-18)-hydroxy fatty acids have been identified in hydrolyzed extracts from marine eustigmatophytes of the genus Nannochloropsis, and C₂₂ to C₂₆ saturated and monounsaturated α-hydroxy fatty acids have been found in three marine chlorophytes. These findings have provided a more complete picture of the lipid distributions within this little studied group of microalgae as well as a range of unusual compounds that might prove useful chemotaxonomic markers. The functions of the hydroxy fatty acids are not known, but a link to the formation of the lipid precursors of highly aliphatic biopolymers is suggested.     

樟科植物中脂肪酸成分的分布

本文报道了樟科11个属49种种子油和‘种果肉油的脂肪酸成分。其中樟族和木姜子族的各属种(滇新樟除外)种子油脂肪酸成分以 C_(10)、C_(12)和 C_(14)饱和或烯酸为主。而厚壳桂属、润楠和楠属种子油脂肪酸成分主要为棕榈酸、油酸和亚油酸。山苍子、天目木姜子、大果山胡椒、山橙、红脉钓樟、鸭公树和锈叶新木姜子种子油中的 C_(10)、C_(12)和 C_(14)烯酸经分离鉴定均是顺式Δ~4烯酸。     

五种库蠓脂肪酸的研究

本文用气相色语法分析了不显库蠓(Culicoides obsoletus)、兴安库蠓(C, sinanoensis)、怒江库蠓(C.Nujiangensis)、灰黑库蠓(C.Pulicaris)和刺整库蠓(C.Punctatus)等5种库蠓雌虫脂 肪醛的组成和含量。 结果表明,5种库蠓均含有12-14种主要脂肪酸,并以棕榈油酸(C_16：1)、棕搁酸 (C_16：0)、油酸(C_18：1)、硬脂酸(C_18：0)含量最高,共占脂肪酸总量的70%以上。对主要脂肪酸的含量进行聚类分析,发现不显库蠓种团内任意两种库蠓间的欧氏距离小于不显库蠓种团种类与灰黑库蠓种团种类间的欧氏距离,说明脂肪酸的定性、定量分析结果对库蠓分类有一定的参考价值。     

EFFECTS OF LOWERING TEMPERATURE DURING CULTURE ON THE PRODUCTION OF POLYUNSATURATED FATTY ACIDS IN THE MARINE DIATOM PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

The composition of fatty acids and contents of eicosapentaenoic acid (EPA) and polyunsaturated fatty acids (PUFAs) of the economically important marine diatom, Phaeodactylum tricornutum (Bohlin), were investigated to see whether reducing the culture temperature enhances the production of EPA and PUFAs. The contents of EPA and PUFAs of P. tricornutum were found to be higher at lower temperature when cultured at 10, 15, 20, or 25°C. When the cells grown at 25°C were shifted to 20, 15, or 10°C, the contents per dry mass of PUFAs and EPA increased to the maximal values in 48, 24, and 12 h, respectively. The highest yields of PUFAs and EPA per unit dry mass (per unit volume of culture) were 4.9% and 2.6% (12.4 and 6.6 mg·L^?1), respectively, when temperature was shifted from 25 to 10°C for 12 h, both being raised by 120% compared with the control. The representative fatty acids in the total fatty acids, when temperature was lowered from 25 to 10°C, decreased proportionally by about 30% in C_16:0 and 20% in C_16:1(n?7) but increased about 85% in EPA. It was concluded that lowering culture temperature of P. tricornutum could significantly raise the yields of EPA and PUFAs.     

海、淡水驯化对5种微藻脂肪酸组成的影响

对5种微藻进行了脂肪酸分析及海淡水驯化影响其脂肪酸组成的研究,结果表明：谈水小球藻和海水小球藻的特征脂肪酸均为16：0、16：2、18：0、18：2和18：3;淡水斜生栅藻的特征脂肪酸为16：0、18：1和18：3;海水三角褐指藻的特征脂肪酸为14：0、16：0、16：1和EPA。淡水藻海水驯化和海水藻淡水驯化后,特征脂肪酸的种类不发生变化,但各种脂肪酸的含量有明显变化,驯化后,几种特征脂肪酸及总脂肪酸占细胞干重的比例在蛋白核小球藻、小球藻－1和三角褐指藻SS02品系中均有不同程度的提高,而在斜生栅藻、小球藻－2和三角褐指藻ZS08、XSO3品系中均有不同程度的下降。     

小果微花藤种仁油主要脂肪酸和甘油三酯组成的研究

小果微花藤(茶茱萸科)[Iodes vitiginea(Hance)Hemsl.](Icacinaccae)种仁含油39-50％,经气相色谱分析,油的主要脂肪酸为十六碳烯酸和棕捅酸。利用臭氧化反应、色谱-质谱和红外光谱方法,鉴定十六碳烯酸为顺-Δ′-十六碳烯酸。用高效液相色谱从油中分离出13个甘油三酯(峰),其中8种甘油三酯进行了色谱鉴定。总甘油三酯中,主要甘油三酯为 HHH(18.4％),HHP(32.3％),PHP(16.8％)。     

REGULATION OF FATTY ACID COMPOSITION BY IRRADIANCE LEVEL IN THE EUSTIGMATOPHYTE NANNOCHLOROPSIS SP.1

Chemical composition and quantitative cytological measurements were determined for the eustigmatophyte Nannochloropsis sp. Cultures were grown in turbidostats at three irradiance levels: growth-limiting light, growth-saturating light and photoinhibiting light. Cellular chlorophyll a content decreased as irradiance level increased, concomitant with a disproportionate reduction in carotenoid content. Nannochloropsis sp. grown in saturating light was characterized by a high content of lipid, fatty acids and carbohydrate compared with cells grown in light-limiting conditions. The increase in cellular lipid content coincided with a reduction in the percentage of eicosapentaenoic acid (C20:5) and arachidonic acid (C20:4), fatty acids that are mainly associated with galactolipids, and with an increase in the relative abundance of palmitic acid (C16:0) and palmitoleic acid (C16:1). At growth limiting light conditions, Nannochloropsis sp. preferentially synthesized galactolipids; however, as growth became light saturated, relatively more neutral lipids, mainly triacylglycerols, were synthesized. Changes in lipid content and composition were qualitatively related to changes in cell morphology. Cells grown under low light conditions were characterized by a large relative volume of chloroplast, high surface density of thylakoid membranes and low relative volume of lipid storage bodies. The physiological implications of the changes in cellular lipid composition and ultrastructure are discussed in relation to light/shade adaptation.