不同氮源对异养小球藻生物量和油脂积累的影响
EFFECTS OF DIFFERENT NITROGEN SOURCES ON GROWTH AND LIPID ACCUMULATION OF A HETEROTROPHIC MICROALGAE-CHORELLA VULGARIS
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摘要: 小球藻因其快速生长和易培养等特性可用于制备生物能源。与传统的光自养相比, 异养小球藻可获得更多的生物量和更高的油脂含量。低成本的马铃薯淀粉水解液可作为小球藻的理想碳源, 在氮饥饿条件下可诱导产生更多的油脂。为了探讨不同氮源对异养小球藻生物量和油脂积累的影响, 并筛选出异养条件下的最适氮源, 实验研究了不同浓度无机氮源NaNO3以及有机氮源丙氨酸和酪氨酸对异养小球藻生物量和油脂积累的影响。以马铃薯淀粉水解液为唯一碳源, 在SE培养基中分别添加不同氮源培养小球藻。设定的NaNO3和丙氨酸浓度均为1.5 mmol /L、3.0 mmol/L、6.0 mmol/L, 酪氨酸浓度为0.75 mmol/L、1.5 mmol /L 和3.0 mmol/L。所有小球藻培养实验均为暗培养并持续10 d时间。实验过程测定的指标为:小球藻的细胞数目、比生长速率、叶绿素含量、中性脂含量和总脂含量。实验结果表明: (1)在异养条件下以硝酸盐为无机氮源时, 氮源促进叶绿素积累从而促进小球藻的生长, 减少硝态氮可以使小球藻快速进入稳定期积累油脂。在NaNO3中氮含量为1.5 mmol/L时, 生物量和油脂含量分别为2.65 g/L和51.21%, 总油脂含量为1.36 g/L。(2)在不添加其他氮源的异养培养基中, 丙氨酸可促进小球藻的生物量增加, 在稳定期仍促进单位细胞的叶绿素含量, 但总油脂含量普遍偏低。(3)酪氨酸可抑制小球藻生物量增加, 使细胞膨大从而促进单位细胞内叶绿素和油脂合成, 油脂含量高达38.78%-47.02%。这些结果表明小球藻可通过诱导氨基酸转运系统适应氮源的变化, 其中酪氨酸所在的第三个转运系统在葡萄糖诱导条件下可促进油脂的合成。Abstract: Chlorella strains have great potential as a resource for production of biofuels due to their relatively fast growth rate and easy cultivation. Heterotrophic cultivation of Chlorella has been developed due to higher biomass concentration and accumulation of much higher lipid content than classical photoautotrophic culture. Instead of using glucose, using low cost materials, such as starch or cellulose-hydrolyzed solution, is a good strategy to reduce the cost of culture medium and the cost of microalgae-based biofuels from heterotrophic fermentation of Chlorella. Potato starch is relatively cheap material in comparison with cornstarch or cassava starch, hence it could be an ideal carbon source for cultivating Chlorella. In a number of oil-producing microalgal species, Chlorella has the capacity of accumulating much higher proportion of fatty acids by nitrogen starvation. The objective of this study was to investigate the effects of various nitrogen resources on cell growth and lipid accumulation in heterotrophic Chlorella vulgaris using potato starch hydrolysate (PSH) as the sole carbon source. In order to illustrate the influence of various nitrogen sources on biomass and lipid content in C. vulgaris, the effects of these nitrogen source including inorganic and organic nitrogen source such as alanine or tyrosine on cell growth and lipid accumulation of C. vulgaris were examined. As the sole carbon source PSH was added to SE medium for algal cultivation prior to addition of all nitrogen sources. NaNO3 and alanine were added to yield starting concentrations of 1.5 mmol /L, 3.0 mmol/L and 6.0 mmol/L, respectively. Tyrosine was added to yield starting concentrations of 0.75 mmol/L, 1.5 mmol /L and 3.0 mmol/L, respectively. Continuous cultivation of C. vulgaris was carried out under dark condition for 10 days. The cell number, special growth rate, cellular chlorophyll content, neutral lipid content and total lipid content of C. vulgaris under present cultural condition were determined throughout the entire experimental period. The results showed that: (1) Higher chlorophyll content and biomass production were observed with higher nitrate-nitrogen concentration, although low concentrations of nitrate-nitrogen can achieve high lipid accumulation in C. vulgaris. The highest biomass productivity was obtained at 1.5 mmol/L sodium nitrate, the biomass of 2.65 g/L, lipid concentration of 51.21% and the total lipid production of 1.36 g/L were achieved. (2) When alanine was used as the sole nitrogen source, it was also observed that the cell growth and total chlorophyll content increased but the lipid content reduced. (3) A low level of biomass production was observed while adding tyrosine as a nitrogen source; however, cell enlargement and high chlorophyll content were achieved under this condition, resulting in high lipid content of 38.78%-47.02%. Based on above results, it was concluded that the unicellular alga C. vulgaris was able to respond to nitrogen nutrient changes by inducing specific amino acid transport systems, and it seemed to have developed more specific mechanisms of amino-acid uptake and lipid production along with subtle regulatory phenomena such as induction by nitrogen starvation. Furthermore, tyrosine, as the part of the third uptake system induced by glucose transport system, can also enhance continually cellular lipid accumulation in C. vulgaris.
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Keywords:
- Chlorella vulgaris /
- Heterotrophic fermentation /
- Nitrogen source /
- Alanine /
- Tyrosine
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