Cazzaniga Stefano, Dall'Osto Luca, Szaub Joanna, Scibilia Luca, Ballottari Matteo, Purton Saul, Bassi Roberto
Dipartimento di Biotecnologie, Università di Verona, Strada Le Grazie, Verona, 15-37134 Italy.
Institute of Structural and Molecular Biology, University College London, London, WC1E 6BT UK.
Biotechnol Biofuels. 2014 Oct 21;7(1):157. doi: 10.1186/s13068-014-0157-z. eCollection 2014.
The utilization of biomass from microalgae for biofuel production is one of the key elements for the development of a sustainable and secure energy supply. Among the different microalgae, Chlorella species are of interest because of their high productivity, high lipid content, and resistance to the high light conditions typical of photobioreactors. However, the economic feasibility of growing algae at an industrial scale is yet to be realized, in part because of biological constraints that limit biomass yield. A key issue is the inefficient use of light due to uneven light distribution, and the dissipation of excess absorbed light as heat. The successful implementation of biofuel production facilities requires the development of algal strains with enhanced light use efficiency in photobioreactors. Such domestication strategies include decreasing the absorption cross section in order to enhance light penetration, increasing the size of metabolic sinks per chlorophyll and minimizing feedback energy dissipation.
In this work we applied random mutagenesis and phenotypic selection to the thermotolerant, fast-growing Chlorella species, C. sorokiniana. Truncated antenna mutants (TAMs) were selected that exhibited a lower fluorescence yield than the wild-type (WT) strain. Six putatively interesting mutants were selected by high throughput fluorescence video imaging, two of which, TAM-2 and TAM-4, were found to have approximately half the chlorophyll content per cell and LHCII complement per PSII with respect to the WT. In batch culture, TAM-2 showed an increased photon use efficiency, yielding a higher Pmax at saturating irradiances with respect to the WT. Cultivation of TAM-2 in both laboratory-scale and outdoor photobioreactors showed higher productivity than WT, with a 30% higher biomass yield in dense cell suspensions typical of industrial photobioreactors.
These results suggest that generation of mutants with low chlorophyll content can significantly improve the light-to-biomass conversion efficiency of C. sorokiniana under mass culture conditions. However, owing to the lack of sexual reproduction in this species, the presence of additional mutations might affect growth rate, suggesting that selection should include evaluation of multiple independent mutants for each desired phenotype.
利用微藻生物质生产生物燃料是实现可持续和安全能源供应发展的关键要素之一。在不同的微藻中,小球藻因其高生产力、高脂质含量以及对光生物反应器典型的高光条件具有抗性而备受关注。然而,藻类在工业规模上生长的经济可行性尚未实现,部分原因是存在限制生物质产量的生物学限制因素。一个关键问题是由于光分布不均匀导致光利用效率低下,以及过量吸收的光以热的形式耗散。生物燃料生产设施的成功实施需要开发在光生物反应器中具有更高光利用效率的藻类菌株。此类驯化策略包括减小吸收截面以增强光穿透,增加每叶绿素的代谢库大小并最小化反馈能量耗散。
在这项工作中,我们对耐热、生长迅速的小球藻——索氏小球藻应用了随机诱变和表型选择。筛选出了截短天线突变体(TAM),其荧光产量低于野生型(WT)菌株。通过高通量荧光视频成像选择了六个假定有趣的突变体,其中两个,即TAM - 2和TAM - 4,相对于WT,其每细胞叶绿素含量和每个光系统II的捕光复合体II(LHCII)含量约为一半。在分批培养中,TAM - 2显示出光子利用效率提高,在饱和辐照度下相对于WT产生更高的最大光合速率(Pmax)。在实验室规模和室外光生物反应器中培养TAM - 2均显示出比WT更高的生产力,在工业光生物反应器典型的密集细胞悬浮液中生物量产量高出30%。
这些结果表明,在大规模培养条件下,产生低叶绿素含量的突变体可显著提高索氏小球藻的光到生物质的转化效率。然而,由于该物种缺乏有性繁殖,额外突变的存在可能会影响生长速率,这表明选择应包括对每个所需表型的多个独立突变体进行评估。