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Functional characterization of a Δ6 fatty acid desaturase gene and its 5′-upstream region cloned from the arachidonic acid-rich microalga Myrmecia incisa Reisigl(Chlorophyta)

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Journal of Oceanology and Limnology 2018年第6期36卷 2308-2321页

作者： ZHANG Li CAO Haisheng NING Pu ZHOU Zhigang Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources Conferred by Ministry of Education Shanghai Ocean University National Demonstration Center for the Experimental Teaching of Fisheries Science Shanghai Ocean University International Research Center for Marine Biosciences Conferred by Ministry of Science and Technology Shanghai Ocean University

It is suggested that Δ6 fatty acid desaturase(FAD) plays a critical role in the biosynthesis of polyunsaturated fatty acids in plants and microalgae. But why does it adapt to the changed environments such as nitrogen starvation is seldom understood. One Δ6 FAD gene( MiD6 fad) from an arachidonic acidrich microalga M yrmecia incisa Reisigl(Chlorophyta) was first heterologously expressed in S accharomyces cerevisiae for the identification of function. The fatty acid profile of transgenic yeast detected by gas chromatography-mass spectrometry illustrated that the enzyme MiD6 FAD could convert linoleic and ?-linolenic acids to γ-linolenic and stearidonic acids, respectively, demonstrating that M iD6 fad encoded a Δ6 FAD. A 1 965-bp fragment of the cloned 2 347-bp 5′-upstream region of M iD6 fad was next subcloned and fused upstream with green fluorescent protein(GFP) gene to replace the GAL1 promoter of the vector pYES2. The generated construct was transformed into S. cerevisiae for function determination. Confocal microscopic images of the transformed line illustrated that this inserted fragment could drive GFP expression, which was further verified by fluorescence intensity quantification and Western blot analysis using antiGFP antibody. The conversion efficiency(approximately 2%-3%) of MiD6 FAD was much lower than the reported ? 3 FAD and Δ6 elongase in this microalga, suggesting that MiD6 FAD catalysed the possible ratelimiting step for ArA biosynthesis. The presence of several putative c is-acting regulatory elements in this identified promoter sheds new light on the regulation mechanism research of Δ6 FAD transcription for the ArA production in M. incisa in changing environmental factors.

关键词： arachidonic acid(ArA) fatty acid desaturase(FAD) green fl uorescent protein(GFP) green microalga Saccharomyces cerevisiae 5′-upstream region(5′-USR)

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Superhydrophobic and superlipophilic biochar produced from microalga torrefaction and modification for upgrading fuel properties

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Biochar 2023年第1期5卷 334-348页

作者： Congyu Zhang Meng Wang Wei-Hsin H.Chen Ying Zhang Anelie Pétrissans Mathieu Pétrissans Shih-Hsin H.Ho State Key Laboratory of Urban Water Resource and Environment School of EnvironmentHarbin Institute of TechnologyHarbinChina School of Resources and Environment Northeast Agricultural University150030 HarbinChina Department of Aeronautics and Astronautics National Cheng Kung University701 TainanTaiwanChina Research Center for Smart Sustainable Circular Economy Tunghai University407 TaichungTaiwanChina Department of Mechanical Engineering National Chin-Yi University of Technology411 TaichungTaiwanChina Universitéde Lorraine INRAELERMAB88000 EpinalFrance

Torrefaction operation is an essential pathway for solid biofuel upgrading,and good hydrophobicity of torrefied biochar is conducive to its ***,a two-stage treatment of torrefaction followed by modification by hexadecyltrimethoxysilane was adopted to improve the moisture resistance performance of *** two-stage treatment process led to a longer torrefied microalgal biochar preservation time(60-200%improved)and great superhydrophobicity and ***,the modified microalgal biochar could significantly adsorb leaking oil for environmental remediation and further improve the calorific value of the *** obtained results indicated that the oil adsorption capacity of modified microalgal biochar was correlated to torrefaction temperature and oil ***,the oil adsorption capacity was enhanced up to 70-80%from the modification process when comparing to raw *** the torrefaction temperature enhanced the adsorption quantity of the modified microalgal *** adsorbing the oil,the calorific value of oilchar,namely,biochar with adsorbed oil,could be higher than 40 MJ kg^(−1).Furthermore,the pyrolysis and combustion characteristics suggested that biochar stability gradually rose as the torrefaction temperature *** comprehensively analyzing and comparing the fuel performance of the modified microalgal biochar with previous literature,the obtained modified microalgal biochar possessed better fuel properties and environmental sustainability.

关键词： microalga Torrefaction Biochar Superhydrophobicity Oil adsorption Fuel property

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EFFECT OF ZINC,SELENIUM AND MANGANESE ON THE GROWTH OF A MARINE microalga NANNOCHLOROPSIS OCULATA(EUSTIGMATOPHYCEAE)

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Chinese Journal of Oceanology and Limnology 2002年第S1期20卷 93-101页

作者：高亚辉杨心宁陈昕王大志洪丽玉 The Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering School of Life. Sciences Xiamen University Xiamen 361005 China MEE Key laboratory of Ministry of Education Research Center for Enviromental Science Xiamen University Xiamen 361005 China

The effect of trace metals of Zinc （Zn）, Selenium （Se） and Manganese （Mn） on thegrowth of a marine microalga was determined by culturing Nannochloropsis oculata in f/2 medium enrichedwith different concentrations of the metal compounds. The results showed that concentration and type ofthe metal compound affected the growth of Nannochloropsis. ZnCl2 and Na2SeO3, promoted the growth ofNannochloropsis at relatively low concentrations of 15. 3 μmol/L and 57. 8 μmol/L respectively, but inhib-it its growth at higher concentration of 153 μmol/L and about 300 μmol/L. No significant growth promotionwas observed for MnCl2 and MnSO4 although MnCl2 was found to have higher toxic effect than MnSO4.

关键词： NannoMoropsis growth metals microalga toxic

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Influences of Natural Colloid in Seawater on the Growth of a microalga

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Journal of Northeast Agricultural University(English Edition) 2002年第2期9卷 131-135页

作者： ZHAO Xin huai,LI Tie jing (College of Food Science,Northeast Agricultural University,Harbin,Heilongjiang,150030,PRC) College of Food Science Northeast Agricultural UniversityHarbinHeilongjiang150030PRC

Colloid concentrates were obtained by employing a cross flow filtration system to ultrafiltrate seawater(which was pre filtrated by 0 45 μm acetate cellulose membrane) sucessively with three *** retentions (colloid concentrates) and control sample(in which colloid was removed)were then inoculated with a microalga and cultivated in *** continuously monitoring cell content during cultivation,it was found that colloid had an obvious beneficial effect on the growth of *** colloids with different sizes influenced microalga's growth in different extent and ways,a brief conclusion could be draw from experiments that colloid might be a potential nutrition source for microalga's growth.

关键词： colloid microalga seawater ultrafiltration

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A method of batch-purifying microalgae with multiple antibiotics at extremely high concentrations

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Chinese Journal of Oceanology and Limnology 2016年第1期34卷 79-85页

作者：韩吉昌王松张琳杨官品赵璐潘克厚 Laboratory of Applied microalgae Biology Key Laboratory of Aquaculture of Ministry of EducationOcean University of China College of Marine Life Sciences Ocean University of China

Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics, alone or in combination, are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials, we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First, microalgal strains of different species or strains were treated with a mixture of ampicillin, gentamycin sulfate, kanamycin, neomycin and streptomycin （each at a concentration of 600 mg/L） for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method, five strains ofNannochloropsis sp. （Eustigmatophyceae）, two strains of Cylindrotheca sp. （Bacillariophyceae）, two strains of Tetraselmis sp. （Chlorodendrophyceae） and one strain ofAmphikrikos sp. （Trebouxiophyceae） were purified successfully. The method shows promise for batch- purifying microalgal cultures.

关键词： antibiotic axenic batch operation bacterium removal microalga

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Predicting the Reproduction Strategies of Several microalgae Through Their Genome Sequences

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Journal of Ocean University of China 2015年第3期14卷 491-502页

作者： GUO Li YANG Guanpin College of Marine Life Sciences Ocean University of China

Documenting the sex and sexual reproduction of the microalgae is very difficult, as most of the results are based on the microscopic observation that can be heavily influenced by genetic, physiological and environmental conditions. Understanding the reproduction strategy of some microalgae is required to breed them in large scale culture industry. Instead of direct observation of sex and sexual reproduction under microscope, the whole set or the majority of core meiosis genes may evidence the sex and sexual reproduction in the unicellular algae, as the meiosis is necessary for maintaining the genomic stability and the advantages of genetic recombination. So far, the available genome sequences and bioinformatic tools （in this study, homolog searching and phylogenetic analysis） allow us to propose that at least 20 core meiosis genes （among them 〉6 must be meiosis specific） are enough for an alga to maintain its sexual reproduction. According to this assumption and the genome sequences, it is possible that sexual reproduction was carried out by Micromonas pusilla and Cyanidiosehyzon merolae, while asexual reproduction was adopted by Bigelowiella natans, Guillardia theta, Nannochloropsis gaditana, N. oeeanica, Chlorella variablis, Phaeodactylum tricornutum and Thalassiosira pseu- donana. This understanding will facilitate the breeding trials of some economic microalgae （e.g., N. gaditana, N. oceanica, C. vari- ablis and P. tricornutum）. However, the reproduction strategies of these microalgae need to be proved by further biological experiments.

关键词： microalga sexual reproduction meiosis core meiosis gene meiosis specific gene homolog searching phylogenetic analysis

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Colloid's influences on microalgae growth as a potential environmental factor

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Chinese Journal of Oceanology and Limnology 2003年第3期21卷 270-279页

作者：赵新淮张正斌刘莲生 Institute of Marine Chemistry

The role of colloid as “colloid pump” in the ocean is well known. The important influence of colloid in seawater on the growth of microalga was found in our 1999—2000 study. Colloid concentrates were obtained by employing a cross flow filtration system to ultrafilter seawater (which had been pre filtrated by 0.45 μm acetate cellulose membrane) successively with different membranes. Ultrafiltration retentions (we called them colloid concentrates) together with control sample (seawater without colloid) were then inoculated with two species of microalgae and cultivated in selected conditions. Monitoring of microalgae growth during cultivation showed that all colloid concentrates had obvious influence on the growth of the microalgae studied. Addition of Fe(OH) 3 colloid or organic colloid (protein or carbohydrate) to the control sample enhanced the microalgae’s growth.

关键词： colloid growth microalga organic colloid seawater ultrafiltration

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Growth Characteristic of the Oleaginous Mi-croalga <i>Chlorella ellipsoidea</i>SD-0701 with Lipid Accumulation

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Natural Resources 2015年第2期6卷 130-139页

作者： Wenyu Luo Wenya Du Yi Su Jiejie Hui Jing Zhuang Lili Liu College of Life Science Tianjin Normal University Tianjin China Tianjin Key Laboratory of Animal and Plant Resistance Tianjin China

microalgae have great advantages as a new biomass source for fuel production. But microalgae are photosynthetic microorganisms, which normally grow in the light. Because of this growth condition, the commercial viability of microalgal biofuel is limited by current production systems. To obtain microalgal biofuel, fermentation is a more convenient, more economical and practical industry model. In this study, we asked whether and why the dark fermentation of C. ellipsoidea SD-0701 could be achieved by changing the culture medium formula. We focused the research on carbon-containing compounds and the initial pH of media. The results indicated that glucose was the optimum carbon-containing compound, which provided C. ellipsoidea SD-0701 with energy and carbon skeleton for accumulating organic compounds including lipids. When C. ellipsoidea SD-0701 was cultivated in the add-nutrition medium containing glucose, the optimum initial pH for the growth of C. ellipsoidea SD-0701 was pH 7.71. Therefore, if the suitable medium is used, C. ellipsoidea SD-0701 can grow normally in the dark, which is the same condition as the fermenter, and high microalgal biomass (0.50 g·L-1) and lipid yield (232.90 mg·L-1) can be achieved.

关键词： Oleaginous microalga Chlorella ellipsoidea Growth Characteristic Lipid Accumulation

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Large-Scale Production of microalgal Lipids Containing High Levels of Docosahexaenoic Acid upon Fermentation of <i>Aurantiochytrium</i>sp. KRS101

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Food and Nutrition Sciences 2013年第9期4卷 1-5页

作者： Won-Kyung Hong Anna Yu Baek-Rock Oh Jang Min Park Chul Ho Kim Jung-Hoon Sohn Akihiko Kondo Jeong-Woo Seo Applied Microbiology Research Center Bio-Materials Research Institute Korea Research Institute of Bioscience and Biotechnology (KRIBB) Jeongeup South Korea Department of Chemical Science and Engineering Graduate School of Engineering Kobe University Kobe Japan. Systems and Synthetic Biology Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB) Daejeon South Korea

In this study, large-scale production of microalgal lipid containing high levels of docosahexaenoic acid (DHA) by fermentation of Aurantiochytrium sp. KRS101 was performed. The microalgal strain yielded productivity of docosahex-aenoic acid (DHA) productivity of 1.08 and 1.6 g/L/d by fermentation at 300-L and 5000-L scale stirrer-type bioreactor. The productivity was significantly enhanced upto 5.6 g/L/d by fermentation at 6000-L scale airlift-type bioreactor, probably due to the reduced shearing force. The microalgal lipid could be efficiently recovered by safe extraction methods such as ethanol extraction, hot water extraction or supercritical fluid extraction, promising commercial potential of the microalgal DHA-rich lipid in the food and feed industry.

关键词： Aurantiochytrium sp. Heterotrophic microalga Large-Scale Cultivation Lipid Docosahexaenoic Acid

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Fast-dissolving antioxidant nanofibers based on Spirulina protein concentrate and gelatin developed using needleless electrospinning

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Food Bioscience 2022年第3期47卷 1281-1293页

作者： Vahid Mosayebi Milad Fathi Mohammad Shahedi Nafiseh Soltanizadeh Zahra Emam-Djomeh Department of Food Science and Technology College of AgricultureIsfahan University of TechnologyIsfahan***Iran Department of Food Science and Engineering College of Agriculture & Natural ResourcesUniversity of TehranKarajIran

Spirulina is a microalga that is well-known for its high protein content and biological activities directly related to its antioxidant *** objective of this study was to produce fast-dissolving antioxidant nanofibers based on Spirulina protein concentrate (SPC) and gelatin using needleless electrospinning *** effect of mixing ratios of SPC (10% w/w) and gelatin (20% w/w) on the viscosity,electrical conductivity and surface tension of electrospinning solutions as well as diameter and morphology of resulting nanofibers was *** the SPC level in the solution blends resulted in a decrease in apparent viscosity and electrical conductivity and an almost stable trend in surface tension (29.25–32.19 mN/m) that led to diminish of diameter of the *** electron microscopy images showed that SPC/gelatin ratio of 40:60 led to the production of uniform and bead-free nanofibers with a relatively smaller average diameter (208.7 ± 46.5 nm).Atomic force microscopy images indicated mesh-like,fibrillary,and bead-free *** transform infrared spectroscopy verified the formation of composite nanofibers and intermolecular interactions between both proteins.X-ray diffraction and thermal analysis showed higher amorphous structure and stability of produced SPC/gelatin nanofibers in comparison to pure materials which was favorable for formation of stable fast-dissolving *** of DPPH and ABTS radical scavenging activities showed that the antioxidant activity of composite nanofibers significantly improved with increasing SPC mixing ratio (p < 0.05).The dissolution test demonstrated that SPC/gelatin nanofibers can be rapidly dissolved in aqueous medium within 2 ***,the results indicated that the electrospun SPC/gelatin nanofibers could be potentially used for nutraceutical delivery in food and packaging applications under high humidity.

关键词： Antioxidant activity Biopolymers Nanoencapsulation Rheology Spirulina microalga

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