有机碳源和氮源对三角褐指藻生长的影响

研究了8种有机碳源和4种氮源对三角褐指藻(Phaeodactylum tricornutum)生长的影响.结果表明,三角褐指藻具有兼养生长的能力,碳浓度为5mmol/L和50mmol/L时,葡萄糖、果糖、乙酸钠和甘油对其生长有明显的促进作用,乳酸钠和乙醇抑制藻细胞生长,半乳糖和柠檬酸钠对其生长随有机碳浓度而异.甘油、葡萄糖和乙酸钠的适宜浓度范围分别为5 -800mmol/L、100 -400mmol/L和50 -300mmol/L.培养液中分别加入50mmol/L甘油、400mmol/L葡萄糖和150mmol/L乙酸钠,培养第14天,三角褐指藻的最大生物量分别为对照的1.45倍、1.25倍和1.11倍.甘油兼养生长的最优氮源是尿素,适宜浓度范围为0.88-8.8mmol/L.当尿素浓度为4.4mmol/L时,最高生物量可达1.31g/L.     

葡萄糖和乙酸钠对湛江等鞭金藻兼养生长的影响

论文探讨了添加外源性有机碳葡萄糖和乙酸钠对湛江等鞭金藻(Isochrysis zhanjiangensis )生长及胞内总脂百分含量的影响.结果表明:随着葡萄糖和乙酸钠浓度的增加,湛江等鞕金藻的生长和产物积累均表现出先促进后抑制的现象.湛江等鞭金藻兼养利用葡萄糖和乙酸钠促进生长的浓度上限分别为50和30g·L^-1.葡萄糖浓度仅15g·L^-1时,对生物量具有显著促进作用,而乙酸钠浓度2.5～15g·L^-1时,对生物量均具有显著促进作用.葡萄糖对藻细胞内总脂百分含量的促进作用浓度范围是1.0～50g·L^-1,而乙酸钠仅在1.0～15g·L^-1时,表现出显著促进作用.葡萄糖浓度为15g·L^-1时,其生物量、总脂百分含量分别比对照增加了30%、30%.乙酸钠浓度为7.5g·L^-1时,其生物量、总脂百分含量分别比对照增加了30%、30%.添加适量的葡萄糖和乙酸钠能够显著促进湛江等鞭金藻生物量及胞内总脂百分含量,葡萄糖和乙酸钠的最适浓度分别是15g·L^-1和7.5g·L^-1.湛江等鞭金藻利用葡萄糖和乙酸钠进行兼养生长的能力是有限的.     

自养和兼养条件下蛋白核小球藻昼夜节律的响应

【目的】研究自养和兼养两种培养方式对蛋白核小球藻(Chlorella pyrenoidosa)生长、细胞分裂和生化组分积累的影响,探讨人工培养蛋白核小球藻的昼夜节律响应机制和优化技术。【方法】小球藻自养培养采用BG11培养基,兼养培养基在BG11培养基中添加4种不同浓度(1、5、10、20 g/L)的葡萄糖,培养周期为10 d。血球板计数法测定藻细胞浓度,干重法测定藻细胞生物量。显微观察藻细胞大小和分裂情况。脂染色法测定小球藻总脂的含量,藻细胞的叶绿素、蛋白和淀粉分别采用甲醇、氢氧化钠、硝酸钙浸提后通过紫外分光光度法定量测定。【结果】葡萄糖兼养培养对蛋白核小球藻具有显著的促生长效应,最适浓度为10 g/L。10 d收获时,兼养组(10 g/L葡萄糖)藻细胞浓度和干重分别是自养组的2.57倍和6.73倍。分析一昼夜中的藻细胞增殖规律可知,第2天和第5天时自养组中增殖的新生子细胞约有76.00%在黑暗期分裂产生,而兼养组中第2天和第5天光照期的新细胞增殖量占比分别达到40.90%和67.50%。一昼夜内藻细胞大小的迁移动态监测表明,第2天自养组藻细胞的体积变化静息期为8 h,兼养组只有4 h;第5天两组藻细胞大小迁移动态的昼夜节律明显,但兼养组黑暗结束后较大细胞(D6μm)占比显著高于自养组。第8天时,兼养组藻细胞已处于稳定期,总脂和蛋白含量均显著高于自养组,藻细胞总脂和色素含量在一昼夜中相对稳定,但蛋白和淀粉含量分别在光照8 h和12 h左右达到峰值。从第2天开始,对兼养组细胞每天进行2 h光延长,收获时藻细胞浓度和干重分别比对照组提高13%和11%。【结论】葡萄糖兼养培养能大幅提高蛋白核小球藻的生物量。蛋白核小球藻生长增殖与生化组分积累均受昼夜节律调控,自养条件下藻细胞以光照期生长黑暗期增殖为主。兼养培养提高藻细胞生物量的机制在于缩短藻细胞生长静息期,在昼夜节律中加速藻细胞生长并显著提高通过细胞周期检查点的细胞比例,光照期效应尤其明显。藻细胞蛋白和淀粉含量昼夜节律明显,最佳收获时间分别在光照8 h和12 h后。     

有机碳化合物对湛江等鞭金藻生长的影响

为了探讨有机碳化合物对湛江等鞭金藻的营养效应,实验设置了在f/2培养基中添加葡萄糖、乙酸钠、半乳糖、甘油、乙醇、柠檬酸钠和甘氨酸等7种有机碳化合物的处理,测定了湛江等鞭金藻(Isochrysis zhanjiangensis)的生长情况。结果表明,参试的7种有机碳化合物中,甘氨酸对湛江等鞭金藻细胞生长的促进作用最明显,而乙醇对藻细胞生长的促进效果不明显,其他5种均有不同程度的促进作用。7种有机碳对湛江等鞭金藻胞内蛋白质含量和总脂的积累量具有一定差异性影响。0.5～10g·L^-1的葡萄糖、乙酸钠均可提高胞内蛋白质和总脂的含量。半乳糖对总脂积累量的影响不明显。     

三角褐指藻甘油酯对氮胁迫的响应

【背景】三角褐指藻作为生物燃料潜在的生产者,在胁迫条件下能通过改变其甘油酯组成来适应外部环境的变化,同时伴随着生物燃料原料甘油三酯(TAG)的积累,研究三角褐指藻甘油酯对氮胁迫的响应机制有利于深入认识TAG的积累过程。【目的】通过分析三角褐指藻在正常和氮胁迫条件下各类脂质含量及其脂肪酸成分的变化,揭示氮胁迫诱导积累的TAG酰基主要来源,以及在胁迫前生成的各极性甘油酯脂肪酸的去向,从而为进一步认识三角褐指藻对氮胁迫的响应机制提供新信息。【方法】利用高效薄层色谱结合气相色谱法分析三角褐指藻在正常和氮胁迫条件下的脂肪酸及甘油酯组分的变化。【结果】三角褐指藻在氮胁迫条件下TAG含量增加至57.8 mg/g时,总甘油酯含量几乎不变,但各甘油酯含量变化差异很大,表现为各极性脂含量显著降低。在此期间,各类甘油酯脂肪酸组成含量的变化表明,三角褐指藻TAG主要积累饱和及单不饱和脂肪酸,即16:0和16:1n7,分别以从头合成及原有极性脂转化为主,极性脂的部分二十碳五烯酸(EPA)作为酰基供体也向TAG发生了转化;此外组成极性脂的多不饱和脂肪酸16:2n4、16:3n4及EPA分解导致其含量显著下降。【结论】当氮胁迫诱导的三角褐指藻TAG含量为57.8 mg/g时,积累的TAG酰基中有48%来自从头合成,52%来自极性脂转化;而氮胁迫诱导所减少的极性脂酰基中有54%转化成TAG,46%发生了分解。     

光发酵强化三角褐指藻生产岩藻黄素

本研究旨在建立光发酵培养三角褐指藻高效生产岩藻黄素的技术体系。在5 L光发酵罐中,系统研究了兼养条件下初始光强、氮源种类和浓度以及光质对于三角褐指藻生物量浓度和岩藻黄素积累的效果。结果表明,在初始光强为100μmol/(m²·s)红蓝(R:B=6:1)混合光、含氮量为0.02 mol/L的胰蛋白胨和尿素混合氮源(1:1, N mol/N mol)优化条件下,三角褐指藻生物量浓度、岩藻黄素含量和产率分别达到了最大值3.80 g/L、13.44 mg/g和4.70 mg/(L·d),比优化前分别提高了1.41、1.33和2.05倍。本研究开发了强化三角褐指藻光发酵生产岩藻黄素的关键技术,促进了海洋天然产物开发。     

不同饵料对卤虫生长、总脂含量及脂肪酸组成的影响

为了提高养殖卤虫的饵料营养价值,了解其不同生长阶段营养成分变化情况,采用单因子试验研究了8种饵料（三角褐指藻、小球藻、微绿球藻、酵母液、三角褐指藻＋小球藻＋微绿球藻、三角褐指藻＋酵母液、小球藻＋酵母液和微绿球藻＋酵母液）对卤虫生长、总脂含量及脂肪酸组成的影响,结果表明：不同饵料种类对卤虫生长、总脂含量及脂肪酸组成的影响显著（P＜0.05）,增长率,以三角褐指藻＋酵母液最优;总脂含量、以三角褐指藻最优（19.67%）,除酵母液外,与其它饵料相差不显著（P＞0.05）;脂肪酸组成效果,以微绿球藻组最优（EPA：18.01%,DNA：0.55%,（n-3）HUFA：19.08%）,与三角褐指藻组相差不大（P＞0.05）,显著高于其它各组（P＞0.05）.同时以三角褐指藻为饵料,研究了卤虫不同生长阶段（体长2、4、6、8、10 mm）总脂含量、脂肪酸组成变化,结果表明：卤虫体长2～10 mm总脂含量为14.27%～20.93%,随体长的增长降低;EPA、DHA及（n-3）HUFA的含量,均随体长的增长降低,EPA含量为：10.47%～20.77%,DNA含量为：0～0.70%,（n-3）HUFA含量为：10.85%～22.01%.结论认为,卤虫以三角褐指藻或三角褐指藻＋酵母液为饵料培养营养价值最佳,其体长小于6 mm营养价值较佳.     

富营养废水中碳氮磷浓度对布朗葡萄藻总脂和总烃的影响

以经过二次过滤的富营养化鱼塘养殖污水为培养液,添加外源的碳、氮、磷元索,研究了污水中不同的外源无机碳、总氮和总磷浓度对布朗葡萄藻(Botryococcus braunii)生物量、总脂和总烃含量的影响.结果表明:(1)以NaHCO3作为碳源,布朗葡萄藻的生物量和总脂含量在外源无机碳浓度为5～10 mg/L时最高,总烃含量在外源无机碳浓度为15mg/L时最高.(2)以KNO3作为氮源,布朗葡萄藻的生物量在总氮浓度为15mg/L时最高,总脂含量在总氮浓度为2mg/L时最高,总烃含量在总氮浓度为20mg/L时最高.(3)以KH2 PO4作为磷源,布朗葡萄藻生物量在总磷浓度为2mg/L时最高,总脂含量和总烃含量在总磷浓度为1.5 mng/L时最高.     

葡萄糖对小球藻(Chloralla sp. HN08)光合作用和生长的影响

【目的】探讨葡萄糖作为外加碳源对热带海洋小球藻(Chloralla sp.HN08)生物质生产和脂、光合色素、碳水化合物及可溶性蛋白等细胞主要成份含量的影响。【方法】分析比较小球藻HN08在光合自养和兼养(添加10 g/L葡萄糖)2种营养方式下的生长速率、细胞密度、光合放氧速率、油脂相对含量,以及可溶性总糖、淀粉和可溶性蛋白的含量。【结果】结果表明,在光照条件下葡萄糖(10 g/L)能促进小球藻(Chloralla sp.HN08)生长,提高细胞终密度,而异养条件下藻细胞逐渐衰亡。兼养条件下,细胞相对生长速率及细胞终密度分别是自养条件下的6.8倍和1.3倍。兼养藻细胞中可溶性糖、淀粉、油脂含量显著高于(P0.05)光合自养细胞,然而可溶性蛋白质和光合色素含量显著低于(P0.05)光合自养细胞。添加葡萄糖的小球藻液的光饱和点和呼吸速率均高于光自养条件下的细胞,但2种培养条件下藻液的净光合速率无显著差异(P0.05)。【结论】光照条件下,添加葡萄糖可显著提高小球藻HN08相对生长速率和细胞终密度,促进油脂与淀粉的积累。     

培养方式对纤细裸藻脂肪酸与氨基酸含量的影响

以纤细裸藻(Euglena gracilis)为实验对象, 研究了培养方式对纤细裸藻生长、脂肪酸、氨基酸的影响, 并探讨了可能的作用机理。结果表明, 与其他培养方式相比, 光诱导可提高纤细裸藻总脂肪酸、单不饱和脂肪酸(MUFA)和多不饱和脂肪酸(PUFA)含量, 分别为2.69、0.52和1.475 g/100 g; 饱和脂肪酸(SFA)含量由高到低依次为异养组、光诱导组、自养组和兼养组, 其中异养组含量达1.008 g/100 g; 游离氨基酸含量由高到低依次为光诱导组、兼养组、异养组和自养组, 分别为381.57、358.1、330.17和231.1 mg/g; 兼养组必需氨基酸含量最高, 为134.37 mg/g。实验结果说明光诱导培养可显著提高纤细裸藻总脂肪酸、MUFA和PUFA含量(P<0.05); 异养培养可显著提高纤细裸藻饱和脂肪酸含量(P<0.05); 兼养培养可显著提高纤细裸藻必需氨基酸含量(P<0.05)。研究结果为阐明纤细裸藻对不同培养方式的响应提供了科学依据, 同时为其开发应用提供数据支持。     

碳源对粉核油球藻生长和脂肪酸组成特性的影响

研究了不同碳源类型（CO2、NaHCO3和葡萄糖）及其浓度对粉核油球藻（Pinguiococcus pyrenoidosus CCMP 2078）生长及脂肪酸组成的影响。结果表明：（1）培养液中适量添加碳源促进了粉核油球藻的生长,三种碳源的适宜添加浓度分别是0.5% CO2,5mmol/L NaHCO3和20g/L葡萄糖,对数生长末期的细胞密度分别是对照的3.10倍、1.47倍和2.78倍;（2）除了低浓度葡萄糖外,其他碳源类型和浓度均降低了TPUFA和EPA占总脂肪酸的比例,提高了TSFA的比例,胞内EPA和TSFA含量均下降;（3）低浓度碳源提高了TSFA和EPA产量。通入0.5% CO2培养的EPA和TSFA产量分别是对照的2.30倍和2.69倍,5mmol/L NaHCO3培养的TSFA产量是对照的1.85倍,5g/L和10g/L葡萄糖培养的EPA和TSFA产量最高分别可达对照的2.11倍和1.58倍。因此,通入低浓度CO2最有利于粉核油球藻的生长以及EPA和饱和脂肪酸的生产,EPA和饱和脂肪酸含量的提高主要是通过生物量的增大来实现的。     

Enhanced biomass and gamma-linolenic acid production of mutant strain Arthrospira platensis

A mutant of Arthrospira platensis PCC 9108, strain M9108, obtained by mutagenesis with UV treatment, was able to mixotrophically grow in an SOT medium containing 40 g of glucose/l. The biomass and specific growth rate of strain M9108 (4.10 g/l and 0.70/d) were 1.9-fold and 1.4-fold higher, respectively, than those of the wild type (2.21 g/l and 0.58/d) under mixotrophic culture condition. In addition, when compared with the wild type, the content of gamma- linolenic acid (GLA) in the mutant was increased when glucose concentration was increased. Compared with the wild type, the GLA content of the mutant was 2-fold higher in autotrophic culture and about 3-fold higher in mixotrophic culture. Thus, the mutant appears to possess more efficient facility to assimilate and metabolize glucose and to produce more GLA than its wild-type strain.     

Pilot-plant-scale outdoor mixotrophic cultures of Phaeodactylum tricornutum using glycerol in vertical bubble column and airlift photobioreactors: studies in fed-batch mode

Pilot-scale (0.19 m column diameter, 2 m tall, and 60 L working volume) outdoor vertical bubble column (BC) and airlift photobioreactors (a split-cylinder (SC) and a draft-tube airlift device (DT)) were compared for fed-batch mixotrophic culture of the microalga Phaeodactylum tricornutum UTEX 640. The cultures were started photoautotrophically until the onset of a quasi-steady-state biomass concentration of 3.4 g L(-)(1). After this, the cultures were supplemented with organic nutrient (glycerol 0.1 M) and a reduced nitrogen source, resulting in an immediate growth rate boost, which was repeated with successive additions of nutrients in all three photobioreactors. During this period the biomass productivity was enhanced compared to photoautotrophic cultures in the three reactors, although differences were found among them. These could be attributed to the different hydrodynamic behavior influencing the transport phenomena inside the cultures. A 25.4 g L(-)(1) maximum biomass concentration was attained in the SC. Further additions of nutrients did not promote any more growth. The consumption of glycerol was quantitative in the first additions but slowed at high biomass concentration, suggesting that a minimum amount of light is needed to sustain growth. No significant effect of the supplied organic nutrient on carotenoids and chlorophylls content was observed, although it had a profound effect on the fatty acid composition. Eicosapentaenoic acid (EPA) content was increased up to 3% (DW) in mixotrophic growth, giving a productivity of 56 mg L(-)(1) d(-)(1), a significant increase compared to the photoautotrophic control, which yielded a maximum EPA content of 1.9% (DW) and a productivity of 18 mg L(-)(1) d(-)(1). The maximum biomass and EPA volumetric yields obtained in this work are comparable with those reported for commercial photoautotrophic monoculture of large quantities of P. tricornutum in closed continuous-run tubular loop bioreactors with tubes that are typically less than 0.08 m in diameter. When the comparison is established in terms of productivities based on the land area occupied, the vertical airlift and bubble-column bioreactors are extraordinarily more productive.     

A two-stage cultivation process for the growth enhancement of Chlorella vulgaris

This study proposes a two-stage cultivation process with an autotrophic growth followed by a mixotrophic process. The results indicated that a two-stage cultivation process using a daily dose of 3 g/L of glucose could achieve 7.4 g/L of biomass, which was about a 64 % increase over simple autotrophic cultivation. In the second stage of mixotrophic cultivation, glucose was regarded as a better carbon source for cell growth, than was glycerol. Linoleic acid (C18:2) would be the primary component in the two-stage cultivation as in the autotrophic cultivation. Even carbon source was provided in the second stage of mixotrophic cultivation; lower light intensity limited the mixotrophic growth, which indicated that photosynthesis still plays an important role in the second stage of mixotrophical cultivation. The final biomass was higher after this two-stage cultivation process, which made it suitable for application in the production scale-up of algal biomass.     

鱼腥藻1017株的混合营养型生长

鱼腥藻１０１７株混合营养型生长有其特点,外源葡萄糖对生长的刺激不仅在低光强（８００ｌｘ）,而且在高光强（７０００ｌｘ）也表现出来。其混合营养型生长速率在８００－７０００ｌｘ范围内随光照强度的增加而增加,在葡萄糖浓度５－２０ｍｍｏｌ／Ｌ范围内随外源葡萄糖浓度增加而增加。鱼腥藻１０１７株混合营养型生长与光能自养生长相比,生长速率明显提高,对数生长期延长,收获物浓度显著增高,生物量显著提高。     

Mixotrophic growth of Phaeodactylum tricornutum on glycerol: growth rate and fatty acid profile

Mixotrophic growth of the eicosapentaenoic acid (EPA)producing diatom Phaeodactylum tricornutum UTEX640 was carried out in 1-L batch cultures under anexternal irradiance of 165 mol photons m^-2s^-1 by supplementing the inorganic culture mediumwith glycerol. The effect on the growth and the fattyacid profile was studied for different initialglycerol concentrations (0–0.1 M). The optimalglycerol concentration was 0.1 M.A lag phase was observed at high glycerolconcentrations. The present study also shows thatsuccessive additions of glycerol at 0.1M concentrationand using ammonium chloride as a nitrogen sourceremarkably increased the maximum biomass concentration(16.2 g L^-1) and maximum biomass productivity(61.5 mg L^-1 h^-1). These values wererespectively 9 and 8-fold higher than in thephotoautotrophically grown control. The level ofsaponifiable lipids in mixotrophically cultured cellswas significantly higher than in photoautotrophicallycultured cells and increased with the glycerolconcentration in the medium. The concentration ofstorage lipids, saturated and monounsaturated fattyacids, were enhanced but the EPA content did notchange significantly. The EPA content was around 2.2%of biomass dry weight. The maximum EPA yield was33.5 mg L^-1 d^-1 and was obtained in aculture containing 0.1 M glycerol, supplementedperiodically by ammonium chloride. This productivitywas 10-fold higher than the EPA productivity obtainedunder mixotrophic conditions.     

Growth characteristics of the cyanobacterium Nostoc flagelliforme in photoautotrophic, mixotrophic and heterotrophic cultivation

Nostoc flagelliforme is a terrestrial cyanobacterium with high economic value. Dissociated cells separated from a natural colony of N. flagelliforme were cultivated for 7 days under either phototrophic, mixotrophic or heterotrophic culture conditions. The highest biomass, 1.67 g L⁻¹ cell concentration, was obtained under mixotrophic culture, representing 4.98 and 2.28 times the biomass obtained in phototrophic and heterotrophic cultures, respectively. The biomass in mixotrophic culture was not the sum as that in photoautotrophic and heterotrophic cultures. During the first 4 days of culture, the cell concentration in mixotrophic culture was lower than the sum of those in photoautotrophic and heterotrophic cultures. However, from the 5th day, the cell concentration in mixotrophic culture surpassed the sum of those obtained from the other two trophic modes. Although the inhibitor of photosynthetic electron transport DCMU [3-(3,4-dichlorophenyl)-1,1-dimethylurea] efficiently inhibited autotrophic growth of N. flagelliforme cells, under mixotrophic culture they could grow by using glucose. The addition of glucose changed the response of N.flagelliforme cells to light. The maximal photosynthetic rate, dark respiration rate and light compensation point in mixotrophic culture were higher than those in photoautotrophic cultures. These results suggest that photoautotrophic (photosynthesis) and heterotrophic (oxidative metabolism of glucose) growth interact in mixotrophic growth of N. flagelliforme cells.     

Effect of organic carbon sources and environmental factors on cell growth and lipid content of Pavlova lutheri

The present study aimed to investigate the effects of organic carbon sources, cultivation methods, and environmental factors on growth and lipid content of Pavlova lutheri for biodiesel production. In the 250-mL flask bioreactors, P. lutheri was cultivated in the modified artificial seawater (ASW) medium containing glucose, glycerol, sodium acetate, or sucrose as an organic carbon substrate. The effects of different growth conditions (phototrophic, mixotrophic, and heterotrophic) and environmental factors such as photoperiod, light intensity, and salinity were evaluated. Growth of P. lutheri was inhibited under heterotrophy but was enhanced in mixotrophy as compared to phototrophy. Biomass and lipid content of P. lutheri were significantly (p < 0.05) affected by changing photoperiod, light intensity, and salinity. Higher biomass concentration and lipid content were observed at a light intensity of 100 ± 2 μmol photons m−2 s−1, 18 h photoperiod, and 30% salinity, in a modified ASW medium supplemented with 10 mmol sucrose. An increase in biomass concentration from 320 ± 25.53 to 1106 ± 18.52 mg L−1 and high lipid content of 31.11 ± 1.65% (w/w) were observed with the optimized culture conditions, demonstrating a significant (p < 0.05) enhancement in biomass and lipid content due to the improved culture conditions. The present study emphasizes the possible use of sucrose for biomass and lipid production with P. lutheri under the optimized culture conditions. Using low-cost and relatively easy accessible feedstock such as sucrose would be a valuable alternative for growing microalgae with enhanced lipid content.     

响应面法优化普通小球藻混合营养培养基组成生产生物质

利用响应面法优化了混合营养培养普通小球藻生产生物质的培养基组成.首先采用Plackett-Burman设计对11个相关营养因素的效应进行了评价,并筛选出影响小球藻细胞生长的3个主要因素为KNO3、葡萄糖和NaC1;然后结合Box-Behnken设计建立了以小球藻浓度为响应值的二次回归方程模型,获得优化的培养基组成为KNO31.64g/L、葡萄糖45g/L、NaC1 1.57g/L;模型预测的最大浓度为5.28g/L,验证值为5.68g/L;验证结果表明,所建立模型预测精度较好,可用于优化小球藻的混养培养基组成.优化条件下混养小球藻细胞的蛋白质和色素含量较优化前降低,而可溶性糖和油脂含量提高,脂肪酸以棕榈酸和油酸为主;细胞组分分析结果显示,混养培养所得小球藻生物质具有作为生产微藻生物能源原料的潜力.