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赵艳, 汪成, 王超霞, 王天圻. 一株新型自絮凝凯式拟小球藻的两步培养产油技术[J]. 水生生物学报, 2019, 43(1): 196-204. DOI: 10.7541/2019.024
引用本文: 赵艳, 汪成, 王超霞, 王天圻. 一株新型自絮凝凯式拟小球藻的两步培养产油技术[J]. 水生生物学报, 2019, 43(1): 196-204. DOI: 10.7541/2019.024
ZHAO Yan, WANG Cheng, WANG Chao-Xia, WANG Tian-Qi. A NEW TWO-STEP METHOD ON CULTURING SPONTANEOUSLY FLOCCULATING MICROALGA PARACHLORELLA KESSLERI[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(1): 196-204. DOI: 10.7541/2019.024
Citation: ZHAO Yan, WANG Cheng, WANG Chao-Xia, WANG Tian-Qi. A NEW TWO-STEP METHOD ON CULTURING SPONTANEOUSLY FLOCCULATING MICROALGA PARACHLORELLA KESSLERI[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(1): 196-204. DOI: 10.7541/2019.024

一株新型自絮凝凯式拟小球藻的两步培养产油技术

A NEW TWO-STEP METHOD ON CULTURING SPONTANEOUSLY FLOCCULATING MICROALGA PARACHLORELLA KESSLERI

  • 摘要: 以分离获得的一株新型自絮凝凯式拟小球藻(Parachlorella kessleri) F01为材料, 自养单步培养法为对照, 设计两步培养法, 研究阶段Ⅰ添加葡萄糖兼养和阶段Ⅱ营养元素限制处理对藻细胞油脂积累及絮凝性能的影响。分别采用血球板计数法、干重法、脂染色法测定藻细胞浓度、生物量和总脂含量, 三维荧光光谱分析藻细胞胞外聚合物(Extracellular polymeric substances, EPS)组分和含量。结果表明: (1)两步培养法阶段Ⅰ兼养培养最适葡萄糖浓度为10 g/L, 10d收获时藻细胞油脂产率204.25 mg/(L·d), 是对照组的16.20倍; 静置12h的藻细胞自絮凝率96.1%, 与对照组差异不显著。(2)在阶段Ⅰ基础上, 阶段Ⅱ进行不同元素限制处理培养1d, 低糖组和低糖低氮协同处理组的藻细胞油脂产率分别为242.64和227.61 mg/(L·d), 分别比阶段Ⅰ增加18.8%和11.4%; 培养4d, 低糖、无糖、低氮和低糖低氮协同4种处理组油脂产量显著高于对照组和阶段Ⅰ, 其中低糖低氮协同处理组最高, 达到3.08 g/L, 是对照组的23.69倍, 比阶段Ⅰ增加了51.0%, 而且阶段Ⅱ中4种处理组藻细胞的自絮凝率基本在85.0%以上, 能满足收获要求。(3) F01藻细胞EPS中蛋白类色氨酸物质含量高低与藻细胞自絮凝率大小密切相关, 不同培养处理通过改变藻细胞EPS中蛋白类色氨酸物质的含量而影响其絮凝性能。自絮凝凯式拟小球藻F01是生物柴油生产的优良潜力藻种, 两步培养法能大幅提升其产油效益。产油微藻的自絮凝优势与两步培养法结合, 有望成为解决微藻生物柴油生产技术瓶颈的关键突破口。

     

    Abstract: A newly isolated spontaneously flocculating microalgae Parachlorella kessleri F01 was used as the material, and the algae sample in autotrophic cultivation of one-step culture method was used as the control group. A two-step culture method was designed in which glucose was supplemented for mixotrophic cultivation at stage Ⅰ, and nutritional limitation treatments were done at stage Ⅱ, and we studied their effects on the lipid accumulation and flocculation performance of the algae. The algae cell number was determined by a hemocytometer. Biomass was measured via dry weight, and the total lipid was quantified with staining method. The extracellular polymeric substances (EPS) components of algae were analyzed by three-dimensional fluorescence spectroscopy. The results showed that: (1) The optimum glucose concentration was 10 g/L at stage Ⅰ. The alga lipid productivity reached to 204.25 mg/L/d on the 10th harvesting day, which was 16.20 times of that in the control group, and the self-flocculation rate of alga cell was 96.1% after sitting for 12h, which has no significant difference with that in the control group. (2) On the basis of stage Ⅰ, alga cells were cultured for 1 day in stage Ⅱ with different elemental treatments. The alga lipid productivity were 242.64 mg/L/d and 227.61 mg/L/d in the low-glucose group and the low-glucose coupled with low-nitrogen group, respectively, increasing by 18.8% and 11.4% of those at stage Ⅰ. After 4 days cultivation, the alga lipid productions of the four treatment groups including low-glucose, no-glucose, low-nitrogen and the low-glucose coupled with low-nitrogen significantly increased compared with those in the control group and stage Ⅰ. The highest production reached to 3.08 g/L in the low-glucose and low-nitrogen co-treatment group, which was 23.69 times of that in the control group, and increased by 51.0% compared with that in stage Ⅰ. The self-flocculation rates in four treatment groups at stage Ⅱ were nearly above 85.0%, which could satisfy the harvesting requirements. (3) The content of protein tryptophan in alga EPS was positively related to the self-flocculation rate. Different culture treatments could change the protein tryptophan content in EPS of algae cells, and thus affect their flocculation performance. Self-flocculation Parachlorella kessleri F01 is an excellent potential alga for biodiesel production. The two-step culture method can greatly enhance its lipid production. The advantage of self-flocculation combined with two-step culture method is expected to be the key breakthrough to solve the bottleneck of biodiesel production technology of microalgae.

     

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