一株葡萄藻的分离、鉴定及产烃性能评价

陈传红, 吴洪, 尹顺吉, 马建源, 李青

陈传红, 吴洪, 尹顺吉, 马建源, 李青. 一株葡萄藻的分离、鉴定及产烃性能评价[J]. 水生生物学报, 2016, 40(5): 1012-1019. DOI: 10.7541/2016.131
引用本文: 陈传红, 吴洪, 尹顺吉, 马建源, 李青. 一株葡萄藻的分离、鉴定及产烃性能评价[J]. 水生生物学报, 2016, 40(5): 1012-1019. DOI: 10.7541/2016.131
CHEN Chuan-Hong, WU Hong, YIN Shun-Ji, MA Jian-Yuan, LI Qing. ISOLATION, IDENTIFICATION AND EVALUATION OF HYDROCARBON PRODUCING PERFORMANCE OF ONE BOTRYOCOCCUS STRAIN[J]. ACTA HYDROBIOLOGICA SINICA, 2016, 40(5): 1012-1019. DOI: 10.7541/2016.131
Citation: CHEN Chuan-Hong, WU Hong, YIN Shun-Ji, MA Jian-Yuan, LI Qing. ISOLATION, IDENTIFICATION AND EVALUATION OF HYDROCARBON PRODUCING PERFORMANCE OF ONE BOTRYOCOCCUS STRAIN[J]. ACTA HYDROBIOLOGICA SINICA, 2016, 40(5): 1012-1019. DOI: 10.7541/2016.131

一株葡萄藻的分离、鉴定及产烃性能评价

基金项目: 

国家973项目(2012CB723606);国家重大科学仪器设备开发专项(2012YQ15008715)资助

ISOLATION, IDENTIFICATION AND EVALUATION OF HYDROCARBON PRODUCING PERFORMANCE OF ONE BOTRYOCOCCUS STRAIN

Funds: 

Supported by the National Basic Research Program of China (973 Program) (2012CB723606)

  • 摘要: 研究从我国海南博鳌海边的淡水池塘水样分离获得一株绿藻ENN41。显微形态观察表明,ENN41的形态特征属于葡萄藻。进一步克隆ENN41的核糖体小亚基18S rRNA片段,利用分子生物学软件进行比对分析,结果表明ENN41的18S rRNA基因序列与布朗葡萄藻(Botryococcus braunii)同源性最高,说明ENN41为一株布朗葡萄藻(B.braunii)。ENN41在柱式反应器中培养16d,单位体积产率为0.483 g/(Ld),粗烃占干重的含量为56.6%;主要脂肪酸为油酸(C18:1)、十八碳四烯酸(C18:4)和棕榈酸(C16:0),三者之和占总脂肪酸的72.6%;利用尼罗红染色,清晰可见大量的油脂分布在细胞内和胞外基质中。ENN41在板式反应器中培养16d,单位体积产率为0.234 g/(Ld),粗烃占干重的含量为20.0%。上述研究表明,ENN41是具有较高的生长速度和粗烃积累能力的布朗葡萄藻(B.braunii)藻株,具有产业化应用潜力。
    Abstract: ENN41 is a freshwater strain isolated from Bo'Ao coast of Hainan Province. Microscopic observation showed that ENN41 has the unique morphological character of genus Botryococcus. The 18S rRNA gene of ENN41 was cloned for further species confirmation at molecular level. The homologous analysis showed that the 18S rDNA gene of ENN41 was highly homologous to the species of Botryococcus braunii in GenBank. ENN41 achieved a biomass productivity of 0.483 g/(Ld) and crude hydrocarbon content of 56.6% for 16 days culture in column reactor. The main fatty acids were oleic acid (C18:1), octadecatetraenoic acid (C18:4) and palmitic aid (C16:0), accounting for 72.6% of the total fatty acid. Large amount of lipid stained with Nile red in the extracellular matrix. ENN41 reached a high productivity after 16 days culture in plate bioreactor[0.234 g/(Ld)] with 20% crude hydrocarbon. All the results showed that ENN41 is a natural Botryococcus braunii strain with high productivity of biomass and lipids and high potential for industrial application.
  • [1]

    Williams P, Laurens L. Microalgae as biodiesel and biomass feedstocks:review and analysis of the biochemistry and economics[J]. Energy Environmental Science, 2010, 3(5):554-590

    [2]

    Chisti Y. Biodiesel from microalgae[J]. Biotechnology Advances, 2007, 25(3):294-306

    [3]

    Singh J, Gu S. Commercialization potential of microalgae for biofuels production[J]. Renewable Sustainable Energy Reviews, 2010, 14(9):2596-2610

    [4]

    Miao X L, Wu Q Y. Study on preparation of biodiesel from microalgal oil[J]. Acta Energiae Solaris Sinica, 2007, 28(2):219-222[缪晓玲, 吴庆余. 微藻油脂制备生物柴油的研究. 太阳能学报, 2007, 28(2):219-222]

    [5]

    Li L, Zheng L, Zheng M G, et al. Effect on lipid accumulation of marine oleaginous microalgae under CO2 enrichment cultivation[J]. Acta Hydrobiologica Sinica, 2013, 37(6):1013-1019[李林, 郑立, 郑明刚, 等. 富碳培养对海洋富油微藻油脂积累特性的影响. 水生生物学报, 2013, 37(6):1013-1019]

    [6]

    Templier J, Largeau C, Casadevall E. Variations in external and in ternal lipids associated with inhibition of the resistant biopolymer from the a race of Botryococcus braunii[J]. Phytochemistry, 1993, 33(5):1079-1086

    [7]

    Metzger P, Largeau C. Botryococcus braunii:a rich source for hydrocarbons and related ether lipids[J]. Applied Microbiology and Biotechnology, 2005, 66(5):486-496

    [8]

    Motohide I, Masahiro O, Nobuyoshi N, et al. Isolation of herbicide-resistant mutants of Botryococcus braunii[J]. Bioresource Technology, 2012, 109:300-303

    [9]

    Tornabene T G, Holzer G, Peterson S L. Lipid profile of the halophilic alga Dunaliella salina[J]. Biochemical and Biophysical Research Communications, 1980, 96:1349-1356

    [10]

    Wu Q Y, Yin S, Sheng G Y, et al. New discovery for hererotrophic etiolation algae to study on producing hydrocarbon[J]. Science in China (Series B), 1993, 23(4):423-429[吴庆余, 殷实, 盛国英, 等. 异养黄化藻类产烃研究新发现. 中国科学B辑, 1993, 23(4):423-429]

    [11]

    Wirth D, Fischer-Lui I, Boland W, et al. Absolute confi-guration and synthesis of dextro caudoxirene the gamete-releasing and gamete-attracting pheromone of the brown alga Peritha liacaudata Phaeophyceae[J]. Helvetica Chimica Acta, 1992, 75:751-758

    [12]

    Banerjee A, Sharma R, Chisti Y, et al. Botryococcus braunii:a renewable source of hydrocarbons and other chemicals[J]. Critical Reviews in Biotechnology, 2002, 22(3):245-279

    [13]

    Senousy H H, Beakes G W, Hack E. Phylogenetic Placement of Botryococcus braunii (Trebouxiophyceae) and Botryococcus sudeticus isolate UTEX 2629(Chlorophyceae)[J]. Journal of Phycology, 2004, 40(2):412-423

    [14]

    Wang P Y, Mao Y X, Kong F N, et al. Morphological and genetic diversity of Botryococcus braunii[J]. Periodical of Ocean University of China, 2011, 41(5):63-70[王朋云, 茅云翔, 孔凡娜, 等. 丛粒藻形态多样性与遗传多样性研究. 中国海洋大学学报, 2011, 41(5):63-70]

    [15]

    Zhang K, Eiichi K. Effect of light intensity on colony size of microalga Botryococcus braunii in bubble column photobioreactors[J]. Fermentation and Bioengineering, 1998, 86(6):573-576

    [16]

    Oyama H, Kim S, Park E R. Effects of light intensity and nutrients on the growth of Botryococcus sp.[J]. Misaengmul Hakhoechi, 1997, 25(3):339-343

    [17]

    Wang X Y, Xie S H. Effects of several factors on Botryococcus braunii[J]. Microbiology, 1996, 23(5):275-277[王修垣, 谢树华. 几种因子对从粒藻株A的效应. 微生物学通报, 1996, 23(5):275-277]

    [18]

    Banerjee A, Sharma R, Chisti Y. Botryococcus braunii:a renewable source of hydrocarbon and other chemicals[J]. Critical Reviews in Biotechnology, 2002, 22(3):245-279

    [19]

    Wang Y L, Li Q Y, Li A F, et al. Growth and photosynthetic physiological characteristics of four Eustigmatophycean species[J]. Biotechnology, 2014, 24(2):91-95[王元丽, 李其雨, 李爱芬, 等. 4株真眼点藻的生长及光合生理特性. 生物技术, 2014, 24(2):91-95]

    [20]

    Gao B Y, Shen D D, He S S, et al. Integrated the biomass production of oleaginous microalga Scenedesmus acuminatus and dairy wastewater treatment[J]. Rene-wable Energy Resources, 2014, 32(5):673-679[高保燕, 沈丹丹, 何思思, 等. 富油微藻尖状栅藻生物质生产与奶牛场废水处理相结合的效果研究. 可再生能源, 2014, 32(5):673-679]

    [21]

    Kojima E, Zhang K. Growth and hydrocarbon production by microalga Botryococcus braunii in bubble column photobioreactor[J]. Journal of Bioscience and Bioengi-neering, 1999, 87:811-815

    [22]

    An J Y, Sim S J, Lee J S, et al. Hydrocarbon production from secondarily treated piggery wastewater by the green alga Botryococcus braunii[J]. Journal of Applied Phycology, 2003, 15(2-3):185-191

    [23]

    Xu L, Liu C Z, Li H Z. Medium selection of Botryococcus braunii and its metabolic mechanism[J]. Chinese Journal of Bioprocess Engineering, 2010, 8(6):40-45[徐玲, 刘春朝, 李华钟. 布朗葡萄藻的培养基选择及其产物代谢规律. 生物加工过程, 2010, 8(6):40-45]

    [24]

    Rao A R, Dayananda C, Sarada R, et al. Effect of salinity on growth of green alga Botryococcus braunii and its constituents[J]. Bioresource Technology, 2007, 98(3):560-564

    [25]

    Tanoi T, Kawachi M, Watanabe M M.Effects of carbon source on growth and morphology of Botryococcus braunii[J]. Journal of Applied Phycology, 2011, 23:25-33

    [26]

    Tanoi T, Kawachi M, Watanabe M M. Iron and glucose effects on the morphology of Botryococcus braunii with assumption on the colony formation variability[J]. Jour-nal of Applied Phycology, 2014, 26:1-8

    [27]

    Largeau C, Casadevall E, Berkaloff C, et al. Sites of accumulation and composition of hydrocarbons in Botryococcus braunii[J]. Phytochemistry, 1980, 19(6):1043-1051

  • 期刊类型引用(2)

    1. 徐振宇, 刘建华. 布朗葡萄藻转录组研究进展. 基因组学与应用生物学. 2018(07): 3049-3055 . 百度学术
    2. 李贵丽, 韩志萍, 魏晓奕, 周伟, 龚霄, 李积华. 能源微藻油脂积累及油脂含量检测技术研究进展. 广东农业科学. 2018(10): 134-142 . 百度学术

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出版历程
  • 收稿日期:  2015-08-20
  • 修回日期:  2016-01-04
  • 发布日期:  2016-09-24

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