高产油小球藻的筛选及其油脂分析
SCREENING AND LIPID ANALYSES OF HIGH OLEAGINOUS CHLORELLA SPECIES
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摘要: 小球藻广泛分布于各种生境, 特别是淡水环境中, 适应性强。其同化产物主要是淀粉, 但在环境胁迫条件下可显著积累中性脂, 其脂肪酸类型主要为C16 和C18, 适合作为生物柴油的原料。我们从中国部分地区水体中分离纯化到若干株小球藻, 通过薄层层析比较分析了21 株产油小球藻的油脂含量, 筛选到一株三酰基甘油含量较高的藻株Chlorella sp. NMX37N。其适宜生长温区为15—35℃, 在25℃时生长速率最快, 比生长速率为0.53/d, 生长的最适光强为250 μmol photons/(m2·s)。批量培养实验显示, 藻细胞的三酰基甘油含量随培养时间延长而增加, 并在培养的稳定期达到最大值, 此时培养液中氮基本被耗尽。在批量培养条件下培养Chlorella sp. NMX37N 约40d, 藻细胞中总脂含量可达到33%左右, 与此相比通过两步培养方式, 将培养至对数后期(约20d)的藻细胞缺氮处理48h 后, 得到的总脂产率相当。通过两步培养方式可以大大缩短培养时间, 使得该藻细胞快速有效积累油脂。另外, 气相色谱分析显示, 该藻的总脂和三酰基甘油的脂肪酸均以C16∶0 和C18∶2 为主, 占总脂肪酸的70% 以上, 且不含C20 以上的长链脂肪酸, 可以作为优质的生物柴油原料。Abstract: Chlorella species which have strong adaptability distributes widely in various habitats, especially in the freshwater. Their assimilation product is mainly starch, but environmental stress can trigger lipid accumulation within the cells. The predominant fatty acids are C16 and C18 in Chlorella species, which are suitable feedstocks for biodiesel production. A number of Chlorella strains were isolated from fresh water of China, and their triacylglycerol contents were analyzed by thin layer chromatography. Out of 21 oleaginous Chlorella strains, a high oleaginous stain, NMX37N, was identified. It grew at temperatures from 15 to 35 but not at 40 , and the optimum conditions℃ ℃ , at 25℃ and 250 μmol photons/m2·s, it grew with a specific growth rate of 0.53/d. Batch culture experiments showed that the content of triacylglycerols in Chlorella sp. NMX37N increased with time and reached to the highest at the late stationary phase when the nitrate was exhausted. The total lipid content was 33% of dry cell weight when Chlorella sp. NMX37N grew under batch culture conditions for 40d. A two-step culture method, namely, transfer to the nitrogen deficiency condition at the late exponential phase (about 20d), gave a higher total lipid (36%) compared with the batch culture method. The main fatty acids (70%) in total lipids and triacylglycerols of Chlorella sp. NMX37N were C16∶0 and C18∶2. In addition, there were no long-chain fatty acids, such as C20, in Chlorella sp. NMX37N. The results indicated that the triacylglycerols in Chlorella sp. NMX37N may serve as excellent feedstocks for biodiesel production.
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Keywords:
- Chlorella /
- Triacylglycerols /
- Fatty acids /
- Biodiesel
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