富油能源微藻的筛选及产油性能评价

为了筛选具有产油潜力的能源微藻,以实验室保藏的20株淡水和海洋微藻(绿藻门18株,真眼点藻纲1株,硅藻纲1株)为研究对象,利用光径为3 cm柱状光生物反应器通气分批培养,通过测定微藻培养物的生物量和总脂含量等指标,从中筛选生长速度快、生物量和总脂含量高的微藻。结果表明:20株微藻的生物量和总脂含量分别在1.81~7.88g/L和16.0%~55.9% dw(% Dry weight)之间,筛选得到具有产油潜力的微藻9株,分别是栅藻(Scenedesmus sp.)(6.34g/L,55.9% dw)、麻织绿球藻(Chlorococcum tatrense)(5.93g/L,46.9% dw)、眼点拟微绿球藻(Nannochloropsis oculata)(7.88g/L,35.0% dw)、油面绿球藻(Chlorococcum oleofaciens)(5.58g/L,45.9% dw)、多形拟绿球藻(Pseudochlorococcum polymorphum)(6.10g/L,40.0% dw)、八月衣藻(Chlamydomonas augustae)(5.78g/L,40.5% dw)、椭圆小球藻(Chlorella ellipsoidea)(5.56g/L,40.7%dw)、椭圆绿球藻(Chlorococcum ellipsoideum)(5.41g/L,38.0% dw)、雪绿球藻(Chlorococcum nivale)(5.55g/L,36.3% dw),其中最具产业化潜力的微藻为栅藻(Scenedesmus sp.),其总脂收获量和单位体积总脂产率分别为3.5 g/L和218.7mg/L·d。     

耐高温拟微绿球藻诱变株的筛选与培养

[目的]筛选拟微绿球藻耐高温突变株,提高其高温耐受能力,延长夏季养殖时间。[方法]对拟微绿球藻进行3次EMS重复诱变和反复45℃高温处理后,利用200 mmol/L HYP压力进行48孔板和24孔板两轮筛选,得到的优势突变株进行反应器高温培养评价和夏季室外培养验证。[结果]夏季室外培养,突变株HT39最终生物积累量和EPA含量分别达到1.93 g/L和3.62%,比野生株提高48.0%和21.9%。[结论]利用重复诱变和多孔板压力筛选的方法获得的优势突变株HT39夏季室外培养表现出较强的高温耐受性能,为选育重要经济藻株拟微绿球藻耐高温品系提供了方法依据。     

两株钝顶螺旋藻紫外诱变株的特征

采用紫外诱变的方法筛选获得了两株优良的稳定钝顶螺旋藻突变株M1-3和M5-1,与出发株相比,M5-1较粗大,M1-3较细,但长很长,藻体螺旋数超过40;两株突变株的生长速度和光合放氧速率均有显著提高;M1-3的藻蓝蛋白含量高于出发藻株20.2%;突变株的长碳连不饱和脂肪酸含量高于出发藻株,总脂中M1-3含花生四烯酸（20：4）4.93%、M5-1含EPA（20：5）2.49%。两株突变株对NH4^ 和Zn^2 的抗性也发生了改变。     

羊肚菌原生质体诱变筛选高生物量高氨基酸含量菌株

研究影响尖顶羊肚菌（MOrchellaconicaPers、）CGAC－9506原生质体制备、再生因素基础上,首次诱变羊肚菌原生质体,进行高生物量高氨基酸含量菌株选育。原生质体诱变再生株MconicaCGAC－950637生物量比出发株提高7．3％,总氨基酸比出发株提高38％。     

一株高蛋白含量小球藻TX的分离鉴定及其诱变育种

【背景】小球藻由于蛋白含量高、营养丰富,在水产养殖上可直接作为鱼、虾、贝类的优质饵料。【目的】对从养殖环境中分离的小球藻进行诱变,选育生长快、蛋白含量高的突变株,为水产养殖天然饵料生产提供优良藻种资源。【方法】以从养殖环境中筛选的生长相对较快且蛋白含量较高的TX作为出发藻株,对该藻株进行分子鉴定,并对该藻株进行紫外诱变、甲基磺酸乙脂(ethyl methyl sulfonate,EMS)诱变和复合诱变,采用96孔板高通量筛选技术和递进式重复筛选方法选育高生物量、高蛋白突变株。【结果】经18SrRNA基因序列分析,TX鉴定为Chlorella sorokiniana,从540个可能的突变株中筛选到8个遗传稳定且生长较快的突变株,其中H10的总蛋白含量达64.2%,可溶性蛋白含量达0.44g/L,干重达0.72g/L,分别较出发藻株提高3.4%、15.8%和26.2%。【结论】突变株H10蛋白含量高且生长较快,可用于天然饵料生产。     

60Coγ射线诱变选育高产虾青素红发夫酵母突变株

为了筛选高产虾青素红发夫酵母突变株,用不同剂量^60Coγ射线对出发菌株菌液进行反复辐射处理,得到突变株W6-318,并对其生物学特性进行了研究。结果表明不同照射剂量下正突变率为10％—36％,在射线诱变剂量为3．5kGy时诱变效果最佳。最优化条件下突变株生物量、总类胡萝卜素和虾青素产量分别为10．15gL^-1、14．97mgL^-1和12．55mgL^-1,分别比出发菌株提高11．17％、86．39％和101．8％。5L发酵罐放大培养中的生物量、总类胡萝卜素和虾青素产量分别为15．56gL^-1、18．54mgL^-1和14．97mgL^-1。     

14种微藻总脂含量和脂肪酸组成研究

比较分析了14种微藻的总脂含量和脂肪酸组成,结果表明：除小球藻、亚心形扁藻、极微小环藻、微绿球藻外,其他微藻的总脂含量均超过其干重的10％。每一纲的微藻脂肪酸组成都有各自特点,绿藻纲中16：0、16：1(n-7)、18：1(n-9)含量较高,但微绿球藻中16：1(n-7)、20：5(n-3)(EPA)含量远高于其他绿藻;金藻纲中含大量14：0、16：0、18：3(n-3)、22：6(n-3)(DHA);硅藻纲中14：0、16：0、16：1(n-7)、EPA含量较高;黄藻纲的异胶藻富含16：0、16：1(n-7)和EPA。     

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％.     

氮源种类及浓度对眼点拟微绿球藻生长密度、油脂产率和二十碳五烯酸含量的影响

氮源是影响微藻生长和油脂积累的重要因素,文中通过单因素试验比较了NaNO3、CO(NH2)2、NH4Cl、CH3COONH4及其浓度对眼点拟微绿球藻生长密度、生长速率、油脂产率、二十碳五烯酸(EPA)含量的影响。结果表明：NH4+更易被眼点拟微绿球藻利用,能更好地促进微藻生长和油脂积累;氮浓度的增加有利于微藻的生长和藻油脂肪酸的去饱和,但不利于微藻油脂的积累。在实验考察的氮源种类和浓度范围内,CH3COONH4是促进眼点拟微绿球藻生长和油脂积累、EPA生成的适宜氮源,其适宜的浓度为5.29 mmol/L。     

重离子诱变创制高产油微拟球藻新品种

以具有产业化前景的微拟球藻Nannochloropsis oceanica OZ-1为实验材料,利用碳重离子进行诱变育种,采用Imaging-PAM和酶标仪进行大规模筛选,最终获得两株高生长速率微拟球藻突变藻株(HP-1和HP-2),进一步分析显示两株突变藻株(HP-1和HP-2)生物量积累较野生型藻株大幅提高,在18d培养末期生物量分别提高了18％和26％,两株突变藻株油脂产率分别为295 mg/(L·d)和275 mg/(L·d),而野生型藻株为247 mg/(L·d).所获两株突变藻株生长速度快、油脂产率高,较野生型藻株优势明显.     

Development of High‐Level Omega‐3 Eicosapentaenoic Acid (EPA) Production from Phaeodactylum tricornutum

Phaeodactylum tricornutum is a lipid‐rich marine diatom that contains a high level of omega‐3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA). In an effort to reduce costs for large‐scale cultivation of this microalga, this study first established a New BBM medium (0.3 x strength BBM with only 3% of the initial phosphate level) to replace the traditional F/2 medium. Phaeodactylum tricornutum could grow in extremely low phosphate concentrations (25 µM), without compromising the EPA content. In the presence of sea salts, silicate addition was not necessary for high rate growth, high EPA content, or lipid accumulation in this species. Using urea as the sole nitrogen source tended to increase EPA contents per dry biomass (by 24.7%) while not affecting growth performance. The use of sea salts, rather than just sodium chloride, led to significantly improved biomass yields (20% increase) and EPA contents of total fatty acid (46–52% increase), most likely because it supplied sufficient essential elements such as magnesium. A salinity level of 35 led to significantly higher biomass yields compared with 20, but salinity had no significant influence on EPA content. EPA became the dominant fatty acid with average levels of 51.8% of total fatty acids during the exponential growth phase at 20 ppt in New BBM medium with sea salts.     

谷氨酸脱盐废液培养普通小球藻(Chlorella vugaris)C9-JN2010

研究小球藻在谷氨酸脱盐废液中生长,藻细胞组分及废液中营养利用的效果。结果表明:停留时间10 d,小球藻在体积分数为2%脱盐废液中生长最佳,对总糖、总氮及总磷的消耗率分别为99.1%、96.0%和97.0%;细胞干质量、比生长速率和最大生产强度分别为5.60 g/L、0.223 d-1和560.0 mg/(L.d),细胞中粗蛋白质、粗脂肪、粗纤维、粗灰分及总磷含量(以质量百分数计)分别为50.0%±5.0%、10.0%±2.0%、4.0%±0.5%、6.0%±0.5%和0.80%±0.10%。     

Growth of and omega-3 fatty acid production by Phaeodactylum tricornutum under different culture conditions

Detailed studies were carried out on the effects of nitrogen source, phosphate, sodium chloride, growth factors, precursors, CO2, temperature, initial pH, and inoculum size on biomass and eicosapentaenoic acid (EPA) production by Phaeodactylum tricornutum. The EPA content of total fatty acids increased with increasing concentrations of nitrate and urea. Sodium chloride was not required for growth or EPA production. While vitamins B1 and B12 did not affect growth significantly, EPA yield was increased by 65% by B12 supplementation. Maximum EPA production occurred when the air gassing supply was supplemented with 1% CO2. Optimum culture temperature and initial pH for EPA production were 21.5 to 23 degrees C and 7.6, respectively. EPA yields of up to 133 mg/liter of culture were observed. EPA constituted up to 30 to 40% of total fatty acids.     

Growth of and omega-3 fatty acid production by Phaeodactylum tricornutum under different culture conditions.

Detailed studies were carried out on the effects of nitrogen source, phosphate, sodium chloride, growth factors, precursors, CO2, temperature, initial pH, and inoculum size on biomass and eicosapentaenoic acid (EPA) production by Phaeodactylum tricornutum. The EPA content of total fatty acids increased with increasing concentrations of nitrate and urea. Sodium chloride was not required for growth or EPA production. While vitamins B1 and B12 did not affect growth significantly, EPA yield was increased by 65% by B12 supplementation. Maximum EPA production occurred when the air gassing supply was supplemented with 1% CO2. Optimum culture temperature and initial pH for EPA production were 21.5 to 23 degrees C and 7.6, respectively. EPA yields of up to 133 mg/liter of culture were observed. EPA constituted up to 30 to 40% of total fatty acids.     

Improving of lipid productivity of the biodiesel promising green microalga <Emphasis Type="Italic">Chlorella pyrenoidosa</Emphasis> via low-energy ion implantation

High lipid content in microalgae is an essential parameter for adopting of microalgal biomass as a feedstock for biodiesel. Mutation is one approach to obtain desired algal strain with high lipid production. In this study, a mutant strain of Chlorella pyrenoidosa was isolated using 1.5?×?10¹⁵ ions cm^?2 s^?1 of N⁺ ion beam implantation technique, which has been widely used in mutagenesis of agricultural crops. N⁺ implantation slightly improved the growth of the mutant over the corresponding wild strain with significant increase in lipid content (32.4 % higher than the wild strain), which resulted in significant increase in lipid productivity by 35 %. In addition, ion implantation mutagenesis of C. pyrenoidosa resulted in 21.4 % decrease in total saturated fatty acids (SFAs) compared to the wild type, with a noticeable increase in polyunsaturated fatty acids (PUFAs). The increase in PUFAs was due mainly to stimulation of hexadecadienoic acid (C16:2) and octadecadienoic acid (C18:2) production. However, the SFA content of wild and mutant strains was 31.7 and 24.9 % of total fatty acids, respectively, highlighting the oxidative stability of biodiesel produced by both strains according to the European standards. Cultivation of C. pyrenoidosa mutant in selenite enrichment medium for five successive cultivation experiments showed insignificant changes in biomass productivity, lipid content, and lipid productivity alongside the study period, which confirms the genetic stability of the produced mutant. The present study confirmed the feasibility of generation of microalgae mutants with significant high lipid production using ion beam implantation.     

不同年龄段养殖宽体沙鳅肌肉营养成分分析与评价

采用国家标准方法对1、2、3龄宽体沙鳅养殖群体的肌肉营养成分及品质进行了分析比较。结果显示: (1)各年龄组宽体沙鳅肌肉粗蛋白质和粗脂肪含量较高且随着年龄的增长呈增长趋势。其中, 粗蛋白在2龄和3龄组增长不显著(P>0.05), 粗脂肪增长显著(P<0.05)。(2)各年龄组肌肉氨基酸组成相同, 均以鲜味特征氨基酸谷氨酸含量最高, 其次为天门冬氨酸; 第一限制性氨基酸均为缬氨酸; 支链氨基酸与芳香族氨基酸的比值(F值)分别为2.22、2.08和2.12。(3)各年龄组宽体沙鳅脂肪酸在组成和含量上略有差异, 表现为不饱和脂肪酸含量高于饱和脂肪酸含量(P<0.05); 不饱和脂肪酸中单不饱和脂肪酸含量较高(P<0.05); 多不饱和脂肪酸(PUFA)在2龄组最高(P<0.05)且EPA+DHA含量随年龄的增长递增(P<0.05)。综上, 各年龄段宽体沙鳅肌肉营养丰富, 味道鲜美, 以2龄鱼体营养质量最优。     

Seasonal variation in fatty acid composition of silver pomfrets,Pampus argenteus (Euphrasen), in Kuwait waters

Proximate and fatty acid composition of wild silver pomfrets, Pampus argenteus, were studied in Kuwait waters for a period of 1 year (November 2007–October 2008) to see whether there were any seasonal compositional differences between males and females. Ten adults (five males, five females) were sampled each month during (i) Pre‐spawning (March–May), (ii) Spawning (June–August), (iii) Post‐spawning (September–November), and (iv) Winter (December–February). Both sexes had significantly (P < 0.05) higher whole body moisture and lower crude protein and lipid contents in winter compared to the respective males and females sampled in other seasons. However, females had significantly higher (9.1%) lipid content during the pre‐spawning season than females in other seasons (7.0–8.2%). The most abundant fatty acid in whole body lipid in both sexes was C16 followed by C18:1n‐9, which accounted for about 31–35% and 22–24% of total lipids, respectively. Males in the pre‐spawning and spawning seasons had significantly higher total monosaturated fatty acids (MUFA) than males and females in post‐spawning and winter. Males had significantly higher total polyunsaturated fatty acids (PUFA) during post‐spawning seasons than females in pre‐spawning and winter seasons. However, there were no significant differences (P > 0.05) in total saturated fatty acids (SFA), PUFA, EPA (eicosapentaenoic acid), DHA (docosahexaenoic acid) or n‐3/n‐6 ratios between respective males and females in different seasons. Livers in males had significantly (P < 0.05) higher MUFA, SFA, PUFA, EPA and DHA than respective females in all months during the spawning season. Female gonads had significantly (P < 0.05) higher MUFA and PUFA but lower SFA content than males in different months during the spawning season. In contrast to the liver, the gonad DHA content and n‐3/n‐6 ratios in females were significantly higher than in males. The gonads from both sexes contained more than double the amount of EPA present in liver; in the case of DHA this was more than three‐fold higher in female gonads, but not in males. Thus, the presence of higher proportions of PUFA, EPA and DHA in gonads, particularly in eggs of silver pomfret, indicates their need for these fatty acids, which may be used as a guideline for dietary essential n‐3 fatty acid requirements for feed formulation of this species. A higher content of DHA in eggs also indicates the higher requirement for DHA in the broodstock diet of silver pomfret.     

Effect of cell density and irradiance on growth,proximate composition and eicosapentaenoic acid production ofPhaeodactylum tricornutum grown in a tubular photobioreactor

Growth and eicosapentaenoic acid (EPA) productivity of the diatomPhaeodactylum tricornutum grown semicontinuously in a helical tubular photobioreactor were examined under a range of irradiances (approximately 56 to 1712 µmol photons m^-2 s^-1) and cell densities (3 × 10⁶ to 18 × 10⁶ cells mL^-1). Self shading sets the upper limit of operational maximum cell density. Higher irradiance increases this upper limit and also increase the growth rate. Biomass productivity and EPA productivity were enhanced at those cell densities which support the fastest growth rate irrespective of irradiance. The cell protein content increased with increasing irradiance and the carbohydrate and lipid content increased with increasing cell density. EPA productivity was greatest at the highest irradiance. This study shows that biomass productivity and EPA productivity can be maximised by optimising cell density and irradiance, as well as by addition of CO₂.Author for correspondence     

Effects of nitrogen source and concentration on growth rate and fatty acid composition of Ellipsoidion sp. (Eustigmatophyta)

In order to study the effects of different nitrogen source and concentrationon the growth rate and fatty acid composition, a marine microalga Ellipsoidion sp. with a high content of eicosapentaenoic acid (EPA) wascultured in media with different nitrogen sources and concentrations.During the pre-logarithmic phase, the alga grew faster with ammoniumas N source than with nitrate, but the reverse applied during thepost-logarithmic phase. The alga grew poorly in N-free mediumor medium with urea as the sole N source. In the same growth phase,ammonium medium resulted in higher yield of total lipid, but the EPA yielddid not differ significantly different from that using nitrate medium. Themaximum growth rate occurred in medium containing 1.28 mmolL^-1 sodium nitrate, while maximum EPA and total lipid contents werereached at 1.92 mmol L^-1, when EPA accounted for 27.9% totalfatty acids. The growth rate kept stable when NH₄Cl ranged from0.64 to 2.56 mmol L^-1, and the maximum content of total lipidand EPA occurred in the medium with 2.56 mmol L^-1NH₄Cl. The EPA content was higher in the pre- thanpost-logarithmic phase, though the total lipid content was lower. Thehighest EPA content expressed as percent total fatty acid was 27.9% innitrate medium and and 39.0% in ammonium medium.