利用市政污水培养Chlorella vulgaris生产生物柴油

为了考察利用南昌市政污水规模化培养富油微藻生产生物柴油,同时达到净化污水的目的,取南昌市青山湖污水处理厂未经任何处理的市政污水作为普通小球藻(Chlorella vulgaris)生长的培养液。监测了C.vulgaris在市政污水中连续培养10 d的特定生长率、生物质产量以及与之相关的市政污水中氨氮(NH4+-N)、总磷(TP)、化学需氧量(COD)、总悬浮固体(TSS)和挥发性悬浮固体(VSS)的清除情况。实验表明:营养物质的水平显著地影响了C.vulgaris的生长。C.vulgaris的生长率在培养8 d后达到最大,OD680为2.856,总的生物质产量日均最大积累速率为0.01 g/L,油脂含量为干质量的18%,油脂的平均日产量为0.001 g/L。培养10 d内NH4+-N、TP和COD的去除率分别为50.0%、32.1%和26.0%,TSS和VSS的日平均去除速率分别为0.01 g/L和0.006 1 g/L。     

Electrotransformation of Chlorella vulgaris

Using hygromycin B resistance as a marker for selection, we have established the conditions required for the transformation of Chlorella vulgaris. The exponentially grown C. vulgaris cells were transformed by electroporation with plasmid pIG121-Hm, and transformants were selected with hygromycin B at a concentration of 50 μg/ml. Cell extracts prepared from the late-log cultures of the transformants exhibited glucuronidase activities as conferred by the gus gene on pIG121-Hm. The maintenance of plasmid in the algal cells seemed to be transient as many cultures derived from the hygromycin B-resistant colonies gradually lost the hygromycin resistance upon prolonged growth. The result of Southern blotting of the genomic DNAs prepared from transformant cultures exhibiting persistent hygromycin resistance showed that integration of part of the plasmid DNA into the host chromosome had taken place. Received: 19 December 1997 / Revision received: 5 October 1998 / Accepted: 27 October 1998     

硒胁迫对小球藻光合色素含量和生长的影响

50 mg/L的硒胁迫下,小球藻中β-胡萝卜素、类叶黄素、叶绿素(叶绿素a、叶绿素b)的含量先明显增加,之后逐渐下降;而在800 mg/L硒胁迫下,各种光合色素含量均明显下降;小球藻活体细胞在693 nm处出现叶绿素a的吸收峰,其吸收值在硒胁迫后明显减弱;在室温荧光发射光谱中,700 nm处的发射峰随着硒浓度的增加而显著下降;荧光激发光谱表明:硒胁迫使小球藻的能量传递受阻,传递效率降低;藻体中水溶性蛋白含量随着硒胁迫的增强而下降.等离子体原子发射光谱(ICP-AES)研究结果表明:随着硒浓度增加,藻体中Mg2 、Ca2 、K 和Na 含量呈明显降低趋势,而培养液中这些离子的浓度则不断增加.     

普通小球藻对养殖污水脱氮除磷的效果研究

随着我国养殖业的不断发展,养殖污水排放量的日益增加,养殖污水的高氮、磷含量导致水体富营养化问题日趋严重。小球藻是光能自养生物,能有效同化氮、磷,使污水中的氮、磷减少。本研究通过在实验室模拟不同氮、磷含量的养殖污水环境,分析小球藻对氮、磷的去除效果;在此基础上,用小球藻处理某养殖场污水;并联合膨润土与小球藻,探究两者脱氮除磷的协同作用能力及膨润土对小球藻细胞沉降的效果。结果表明,小球藻对模拟污水的氨氮去除率可达80%,对磷酸根的最高去除率接近100%;对养殖污水中的氮、磷也有一定的去除效果;但养殖污水成分复杂,小球藻的生长被抑制。膨润土与小球藻的结合,能够提高污水中的氮磷去除率并帮助藻细胞快速沉降,为污水处理后藻细胞的收集处理提供了有效方法。     

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。     

Heavy-metal-induced proline accumulation and its role in ameliorating metal toxicity in Chlorella vulgaris

Exposure of Chlorella vulgaris to elevated concentrations of copper, chromium, nickel and zinc led to intracellular accumulation of high concentrations of these metals. Concomitantly, accumulation of free proline occurred, depending on the concentration of metals in the external medium or in the cell. The greater the toxicity or accumulation of a metal, the greater the amount of intracellular proline in algal cells. However, higher concentrations of copper and chromium were inhibitory to proline accumulation by the test organism. The accumulation of proline was triggered within a few hours of metal treatment. Test metals also induced lipid peroxidation; copper was the most efficient inducer whereas zinc was the least. Pretreatment of C. vulgaris with proline counteracted metal-induced lipid peroxidation and potassium ion efflux. Thus the present work shows a protective effect of proline on metal toxicity through inhibition of lipid peroxidation.     

Oxidative stress and antioxidant content in Chlorella vulgaris after exposure to ultraviolet-B radiation

Growth of Chlorella vulgaris was measured in cultures irradiated with 0, 0.8, 2.0 and 4.4 kJ m² UV-B. Growth expressed as chlorophyll content, declined significantly with increased UV-B dose. Ultraviolet-B irradiated cultures in log phase of growth showed a 284% increase in oxygen radical generation and a 145% increase in lipid peroxidation compared with unirradiated cultures, whereas cultures in the stationary growth phase showed no significant changes in these parameters. The activities of superoxide dismutase and catalase increased by 40 and 500%, respectively, after exposure to a UV-B dose of 4.4 kJ m⁻². Contents of the lipophilic antioxidants α-tocopherol and β-carotene increased by 180 and 63 amol cell⁻¹ respectively, between log and stationary phases in unirradiated cultures; but in UV-B-irradiated cultures these increases were significantly depressed. Photoreducing capacities of chloroplasts were decreased following UV-B irradiation of both isolated chloroplasts and those isolated from irradiated algae. Cells exposed to UV-B exhibited increased size and starch accumulation. These results suggest that oxidative stress conditions related to UV-B exposure trigger an antioxidant response that includes an increase in the activity of the antioxidant enzymes (superoxide dismutase and catalase).     

热带普通小球藻培养模式的筛选及其培养基的优化

【背景】从海南热带海区中分离得到一株微藻,其生长速度快、适应力强,经鉴定该微藻为普通小球藻。【目的】提高热带普通小球藻的生长速率。【方法】以"宁波大学3#微藻培养液配方"为基础培养液,分别添加有机碳(C6H12O6和CH3COONa)对热带普通小球藻进行自养、兼养及异养培养,获得促进热带普通小球藻快速生长的培养方式。在"宁波大学3#微藻培养液配方"的基础上对热带普通小球藻的兼养培养基配方进行优化,并用优化兼养培养基与"宁波大学3#微藻培养基"对比培养热带普通小球藻。【结果】添加6 g/L CH3COONa的兼养模式促进热带普通小球藻生长效果最好;优化的兼养培养基配方为:6 g/L CH3COONa,20 mg/L(NH4)2SO4-N,5 mg/L Na H2PO4-P,3 mg/L Fe SO4-Fe,1 mg/L Vitamin B1和0.000 5 mg/L Vitamin B12。对比培养实验结果显示,培养第6天,兼养培养液收获的生物量(细胞密度)达4.20×107 cells/m L,是"宁波大学3#配方微藻培养液"的2.30倍。【结论】兼养培养模式为热带普通小球藻的最佳培养模式,优化的兼养培养基极显著地提高了热带普通小球藻的生物量(P0.01)。     

小球藻去除水产加工废水中氨态氮的初步研究

目的：确定小球藻去除水产行业加工废水中氨态氮的优化环境条件,为利用小球藻处理此类废水的规模化发展提供基础的实验数据。方法：在静止摇瓶培养方式下,逐一改变废水与小球藻培养液的混合体系的初始pH、温度或光照强度等条件,定时测定体系的氨态氮含量,并根据其随培养时间的变化确定优化环境条件。结果：当小球藻藻液的初始浓度为1×10~6个/mL左右,且小球藻培养液与水产加工废水的配比为3：2时,小球藻去除氨态氮的优化环境条件为：初始pH 7．5、培养温度25℃、光照时间12h/d,光强5300k,此时氨态氮的去除率达到77%。结论：实验条件下小球藻能够较好地去除水产加工废水中的氨态氮,为小球藻规模化处理此类废水提供了基础数据。     

Strategies to enhance cell growth and achieve high‐level oil production of a Chlorella vulgaris isolate

The autotrophic growth of an oil‐rich indigenous microalgal isolate, identified as Chlorella vulgaris C? C, was promoted by using engineering strategies to obtain the microalgal oil for biodiesel synthesis. Illumination with a light/dark cycle of 14/10 (i.e., 14 h light‐on and 10 h light‐off) resulted in a high overall oil production rate (v_oil) of 9.78 mg/L/day and a high electricity conversion efficiency (E_c) of 23.7 mg cell/kw h. When using a NaHCO₃ concentration of 1,500 mg/L as carbon source, the v_oil and E_c were maximal at 100 mg/L/day and 128 mg/kw h, respectively. A Monod type model was used to describe the microalgal growth kinetics with an estimated maximum specific growth rate (μ_max) of 0.605 day^?1 and a half saturation coefficient (K_s) of 124.9 mg/L. An optimal nitrogen source (KNO₃) concentration of 625 mg/L could further enhance the microalgal biomass and oil production, leading to a nearly 6.19 fold increase in v_oil value. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Oxidative damage to chloroplasts from Chlorella vulgaris exposed to ultraviolet-B radiation

Upon UV-B irradiation, Chlorella vulgaris cells and isolated chloroplasts increased in size and starch accumulation . Photosynthetic capacity and chlorophyll content of chloroplasts isolated from irradiated algae decreased by 72 and 66%, as compared to chloroplasts isolated from control cells. Dihydrorhodamine 123 conversion to rhodamine 123 was used as a sensitive method for detection of peroxide (presumably hydrogen peroxide) formation in isolated chloroplasts. The accumulation of rhodamine 123 is higher in irradiated than in nonirradiated chloroplasts and the increased accumulation of rhodamine 123 depended on the UV-B dose. Quantitation of alkyl radical-EPR signals in chloroplasts indicated that UV-B exposure significantly increased radical content in the membranes. The content of an oxidized DNA base (8-hydroxy-2'-deoxyguanosine) in chloroplasts was increased by 72 and 175% after irradiation of the algal culture with 17.3 and 42.6 kJ m⁻², respectively. The chloroplastic activity of superoxide dismutase decreased by 50% as compared with control values after irradiation with 42.6 kJ m⁻² and no changes in ascorbate peroxidase activity and ascorbic acid content were detected at the irradiation doses tested. The β-carotene content in chloroplasts was not affected by the irradiation, but the α-tocopherol content increased approximately 4-fold after UV-B irradiation. The results suggest that oxidative damage related to UV-B exposure is responsible for alterations in chloroplasts function and integrity, and that an antioxidant response is triggered in chloroplasts through an increase in α-tocopherol content.     

小球藻细胞活性物质的提取及对啤酒酵母的生理效应

小妹藻（Chlorella vulgaris）为绿藻门普生性单细胞藻类。因其具有极为丰富均衡的营养成分,常作为医疗保健品,或用于食品、轻工业以及饲料添加剂,并可用于重金属的生物吸附、污水处理及环境评价方面．小球藻生长因子（Chlorena Growth Factor,CGF）又称小球藻精,为其细胞活性物质,含有氨基酸、核酸、多糖、多肽、蛋白质、酶、维生素、矿物质等成分,被誉为“类荷尔蒙”。     

植物激素在小球藻异养培养中的作用

本文研究了几种植物激素对小球藻异养培养的影响。结果表明 ,IAA、IBA及 6_BA三种植物激素均不同程度地促进了小球藻的异养生长 ,培养≤ 36h时 ,IAA或IBA以 2 0mg/L的促进小球藻异养生长的效应最大 ,1 0 0mg/LIAA或IBA则抑制了藻的生长 ;>36h时 ,1 0 0mg/LIAA或IBA表现出促进小球藻生长的效应 ,并最终获最大净A540 增长量 ;6_BA以 0 1mg/L的促进作用最大。IBA与 6_BA组合同样表现出促进小球藻异养生长的效应 ,但并非IBA和 6_BA简单的加合效应 ,5mg/LIBA与 6_BA组合的效应维持 6_BA单因子的作用趋势 ,2 0mg/LIBA与 1mg/L 6_BA组合的效应大于与 0 .1mg/L 6_BA组合的 ,1 0 0mg/LIBA与 0 .1mg/L 6_BA组合的效应在≤ 36h时大于与 1mg/L 6_BA组合的 ,>36h时则相反。另外 ,高浓度IBA(≥ 2 0mg/L)与 6_BA组合抑制了前中期异养藻对葡萄糖的吸收 ,但加速了中后期葡萄的吸收。再者 ,IBA与 6_BA组合加速了异养小球藻对NO_3 的吸收。     

普通小球藻对嗪草酮、骠马和甲草胺的敏感性研究

通过96h的毒性实验,研究了普通小球藻对3种不同作用机制农田常用除草剂嗪草酮、骠马和甲草胺的敏感性．结果表明。在实验条件下,嗪草酮、甲草胺对普通小球藻的毒性随时间的推移有加重趋势。并呈现很好的剂量效应关系;最高抑制生长浓度(嗪草酮0．24mg·L^-1,甲草胺12．8mg·L^-1)处理组的最大比增长率分别为对照组的12．38％和31．58％;骠马低浓度对普通小球藻的抑制作用不明显,并呈一定的生长促进效应,0．08mg·L^-1浓度组普通小球藻最大比增长率为对照组的111．44％,而高浓度则具有明显的生长抑制作用,并随时间推移,毒性逐渐减弱．嗪草酮、骠马和甲草胺的96hEC50分别为0．021、0．937和5．54mg·L^-1．普通小球藻对嗪草酮最敏感。其次为骠马和甲草胺．3种除草剂在实验条件下对普通小球藻叶绿素a含量的影响和对普通小球藻生长的影响相似。表现出较好的剂量．效应关系．     

pH值对沼液培养的普通小球藻生长及油含量积累的影响

以50％的沼液为普通小球藻的全营养培养基,考察培养基的起始pH值对小球藻生长及油脂含量的影响,普通小球藻对不同初始pH的沼液中氮、磷的去除情况。设定了2组实验,一组只调节初始接种培养液的pH,分别为6.0、6.5、7.0、7.5、8.0、8.5;另一组将培养液pH分别固定在6.0、6.5、7.0、7.5、8.0、8.5,pH用稀HCl和NaOH进行调节。研究发现在pH 6.5和pH 7.0的偏酸环境有利于小球藻生长,而pH在7.0~8.5的偏碱性条件下有利于小球藻油脂的积累,因此综合小球藻生长和油脂积累2个因素,得到最适合小球藻生长和油脂积累的pH为7.0。培养结束后沼液中氮磷的去除率分别达到了95％和97％,沼液中的总氮由原来的134.91 mg/L降至4.86 mg/L,总磷由10.19 mg/L降到0.32 mg/L。     

Carbon dioxide biofixation by Chlorella vulgaris at different CO2 concentrations and light intensities

In order to develop an effective CO₂ mitigation process using microalgae for potential industrial application, the growth and physiological activity of Chlorella vulgaris in photobioreactor cultures were studied. C. vulgaris was grown at two CO₂ concentrations (2 and 13% of CO₂ v/v) and at three incident light intensities (50, 120 and 180 μmol m^?2 s^?1) for 9 days. The measured specific growth rate was similar under all conditions tested but an increase in light intensity and CO₂ concentration affected the biomass and cell concentrations. Although carbon limitation was observed at 2% CO₂, similar cellular composition was measured in both conditions. Light limitation induced a net change in the growth behavior of C. vulgaris. Nitrogen limitation seemed to decrease the nitrogen quota of the cells and rise the intracellular carbon:nitrogen ratio. Exopolysaccharide production per cell appeared to be affected by light intensity. In order to avoid underestimation of the CO₂ biofixation rate of the microalgae, exopolysaccharide production was taken into account. The maximum CO₂ removal rate (0.98 g CO₂ L^?1 d^?1) and the highest biomass concentration (4.14 g DW L^?1) were determined at 13% (v/v) CO₂ and 180 μmol m^?2 s^?1. Our results show that C. vulgaris has a real potential for industrial CO₂ remediation.     

Effect of nanoparticle on cellular growth and lipid production in Chlorella vulgaris culture

Magnetic cobalt ferrite/silica nanoparticles (MSNs) and methyl functionalized MSNs (methyl‐MSNs) were used to enhance lipid production in Chlorella vulgaris culture through enhancement of gas‐water mass transfer and increased dissolved concentration of CO₂. Methyl‐MSNs enhanced CO₂–water mass transfer rate better than MSNs, and 0.3 wt% methyl‐MSNs are more effective than 0.1 wt% MSNs. In the cultivation experiment, 0.3 wt% methyl‐MSNs yielded the highest dry cell weight and subsequently, the highest mass transfer rate. However, enhancement of mass transfer rate did not increase lipid content. The volumetric lipid productivity in C. vulgaris culture depends not only on intracellular lipid content but also on the cell mass concentration. Consequently, 0.1 wt% methyl‐MSNs yielded the highest volumetric lipid productivity in C. vulgaris cultivation. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:929–933, 2018     

不同光质对小球藻光自养培养积累油脂的影响

研究了5种光质对小球藻(Chlorella vulgaris)M209256生长和产油的影响。结果表明:蓝光为小球藻的最适生长和产油光质;与其他光质相比,蓝光培养的小球藻生物量和油脂含量均较高,为2.40×107个/mL和28%;红光培养的小球藻生长最慢且油脂含量最低,为1.32×107个/mL和15.13%,表现出明显的"红降"现象。在GCMS分析的基础上,对油脂甲酯化后的十六烷值进行评估,结果发现:蓝光的十六烷值最高;5种光质培养的小球藻所产油脂,甲酯十六烷值均在47以上。因此,小球藻油脂所制备的生物柴油具有较好的燃烧性能。     

Hydrolysis of Lipid-Extracted Chlorella vulgaris by Simultaneous Use of Solid and Liquid Acids

Microalgal biomass was hydrolyzed using a solid acid catalyst with the aid of liquid acid. The use of solid acid as the main catalyst instead of liquid acid was to omit subsequent neutralization and/or desalination steps, which are commonly required in using the resulting hydrolysates for microbial fermentation. The hydrolysis of 10 g/L of lipid-extracted Chlorella vulgaris containing 12.2% carbohydrates using 7.6 g/L Amberlyst 36 and 0.0075 N nitric acid at 150°C resulted in 1.08 g/L of mono-sugars with a yield of 88.5%. For hydrolysis of higher concentrations of the biomass over 10 g/L, the amount of Amberlyst 36 needed to be increased in proportion to the biomass concentration to maintain similar levels of hydrolysis performance. Increasing the solid acid concentration protected the surface of the solid acid from being severely covered by cell debris during the reaction. A hydrolysate of lipid-extracted C. vulgaris 50 g/L was used, with no post-treatment of desalination, for the cultivation of Klebsiella oxytoca producing 2,3-butanediol. Cell growth in the hydrolysate was found to be almost the same as in the conventional medium with the same monosaccharide composition, confirming its fermentation compatibility. It was noticeable that the yield of 2,3-butanediol with the hydrolysate was observed to be 2.6 times higher than that with the conventional medium. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2729, 2019     

Uptake of cadmium and zinc by the alga Chlorella vulgaris: II. Multi-ion situation

Many microorganisms are capable of sequestering and concentrating heavy metals from their aqueous environment. While much research has beep carried out on the uptake of single species of metal ions, little attention seems to have been given to the study of multimetal ion systems. A mathematical model has previously been developed to describe the uptake of individual metal species by a microorganism. The model proposes two sequential processes: an initial rapid uptake due to cellular surface adsorption and a subsequent slow uptake due to membrane transport of the metal into the cells. This article extends the treatment by considering the uptake of two metal species together, cadmium and zinc, under different experimental conditions. The results are discussed in terms of possible mechanistic interactions.