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引用本文:刘 倩,何梅琳,李 凌,于文俊,徐年军,刘建国.一种能源微藻IOAC689S 的筛选和优化培养[J].海洋科学,2011,35(1):29-35.
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一种能源微藻IOAC689S 的筛选和优化培养
刘 倩1,2, 何梅琳2, 李 凌2, 于文俊2, 徐年军1, 刘建国2
1.宁波大学生命科学与生物工程学院;2.中国科学院海洋研究所
摘要:
利用氯仿-甲醇(2:1)提取和气相色谱技术, 对采集自渤海、黄海、东海、南海海域以及实验室现有的海洋微藻进行了总脂和脂肪酸检测, 从中筛选到一株高产油微藻IOAC689S, 其油脂含量在快速生长阶段达到37.5%, 其中可作为生物柴油开发的18C 以下脂肪酸占总脂肪酸的91.03%。通过对其ITS和5.8S rDNA 全序列进行扩增, 得到了包括18S rDNA 及28S rDNA 部分序列在内的全长为998bp 的DNA 片段, 初步确定为一种绿藻。随后利用氧电极检测藻液净光合放氧速率的方法, 研究了光照强度、温度、盐度、pH 值、碳源、N 和P 浓度等不同理化因子对IOAC689S 光合作用的影响, 从而得到该藻优化后的培养参数如下: 光照强度200 μmol/(m2·s)、光下培养温度30~35℃, 黑暗培养温度24℃、盐度30、pH 值7~9、NaNO3 氮源质量浓度75 mg/L、NaH2PO4 磷源质量浓度40 mg/L, 而NaHCO3 碳源对该藻生长没有明显作用。利用上述优化条件培养该藻获得了较好生长效果, 平均比生长速率为0.210 d?1,每周生物量可增加3.8 倍。实验结果还表明在培养后期当pH 值提高至10~11 时, 藻体易发生沉降, 可以利用该特性进行藻体收获。
关键词:  生物柴油  光合放氧  脂肪酸  优化培养  微藻
DOI:
分类号:
基金项目:国家高技术研究发展计划(863 计划)项目(2008AA09Z403);国家重点基础研究发展计划(973 计划)项目(2011CB200904); 中国科学院知识创新工程重要方向项目(KGCX2-YW-373-3)
A biodiesel producing micro-algal strain IOAC689S and optimization of its culture parameters
Abstract:
Total lipid contents and fatty acid compositions of marine microalgae collected from Bohai, Huanghai, Donghai, and Nanhai area and the ones that already existed in the lab of Institute of Oceanology, Chinese Academy of Sciences were analyzed by chloroform-methanol (2:1) extraction and gas chromatography (GC) technology. Marine microalga listed as IOAC689S was unique and suitable for biodiesel production because of its high total lipid content (37.5%) and its high content (91.03%) of short- chain fatty acids (carbon chain length shorter than 18). Total ITS and 5.8S rDNA sequences of this alga were amplified; and a length of 998 bp DNA fragment including 18S rDNA and 28S rDNA sequences was obtained. The changes of net photosynthetic O2 evolution in IOAC689S under different culture conditions (including light intensity, temperature, pH, salinity, nitrate, phosphate and carbonate) were determined by an oxygen electrode. Optimized culture conditions were established (200 μmol/(m2·s) illumination, 30~35℃ in light and 24℃ in darkness, 30‰ salinity, pH 7~9, 75mg/L of NaNO3, 40 mg/L of NaH2PO4). NaHCO2 as carbon-source showed no significant effect on net photosynthetic O2 evolution. Under the optimized culture conditions, 3.8 times biomass increase per week and 0.210 d?1 of average specific growth rate were observed for IOAC689S. In addition, the alga easily flocculated at the late cell culture stages when the culture pH was gradually adjusted to 10 or above. This flocculation can be used for biomass harvesting.
Key words:  Bio-diesel  photosynthetic O2 evolution  fatty acid  optimization  micro algae
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