Efficient heterotrophic cultivation of Chlamydomonas reinhardtii
Efficient heterotrophic cultivation of Chlamydomonas reinhardtii
复制标题
莱茵衣藻高效异养培养
DOI：
10.1007/s10811-018-1666-0
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发表时间：
2019-06-01
影响因子：
3.3
通讯作者：
Li, Yuanguang
中科院分区：
文献类型：
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作者：
Zhang, Zhen;Tan, Yingying;Li, Yuanguang
莱茵衣藻具有独特的细胞结构和清晰的遗传背景，已成为微藻和低等植物基础研究的模式物种。莱茵锦鸡儿的细胞核、叶绿体和线粒体基因组的遗传转化已经成功开展。然而，对莱茵哈迪尔乳杆菌的研究主要集中在生理学和分子遗传学方面，对其培养方法的研究较少。传统的光自养和混合营养培养规模小、生长慢、产量低，严重限制了莱茵梭菌的进一步发展和应用。本研究旨在优化和放大莱茵哈迪尔乳杆菌异养培养工艺。优化的摇瓶培养条件为：装液量150mL，L−1初始接种量0.08g，生长温度30℃，摇床转速150rpm。此外，本研究还证实，虽然莱茵梭菌能够同化异养生长的乙酸(最适初始浓度为80 mM)，但葡萄糖和甘油都不是唯一的碳源。硝酸盐、氨和尿素均可作为异养培养的氮源，其最佳碳氮比为40：1。在最佳条件下，莱茵曲霉的生物量从0.457提高到L−1的1.32g。此外，首次将莱茵曲霉的异养培养扩大到5-L和50-L发酵罐，其中L−1的细胞密度在237h后达到25.44g，是文献报道的2.82倍。
With a unique cell structure and clear genetic background, Chlamydomonas reinhardtii has become a model species for fundamental research on microalgae and lower plants. Genetic transformations in the nucleus and chloroplast and mitochondrial genomes of C. reinhardtii have been successfully developed. However, research of C. reinhardtii has mainly focused on physiology and molecular genetics and few studies investigating cultivation methods have been performed. Small scales, slow growth, and low yields from conventional photoautotrophic and mixotrophic cultures severely limit the further development and application of C. reinhardtii. This study sought to optimize and scale up the heterotrophic cultivation process for C. reinhardtii. Critical parameters in 500-mL shake flasks were optimized as follows: liquid volume of 150 mL, initial inoculation of 0.08 g L−1, growth temperature of 30 °C, and shaking speed of 150 rpm. In addition, this study confirmed that, although C. reinhardtii can assimilate acetic acid for heterotrophic growth (optimum initial concentration of 80 mM), neither glucose nor glycerol can be the only carbon source. Nitrate, ammonia, and urea can be used as nitrogen sources for heterotrophic culture, while the optimal carbon and nitrogen ratio for heterotrophic culture was 40:1. Under optimal conditions, the C. reinhardtii biomass increased from 0.457 to 1.32 g L−1. Furthermore, for the first time, heterotrophic cultures of C. reinhardtii were scaled up to 5-L and 50-L fermenters, in which the cell density reached 25.44 g L−1 after 237 h, which is 2.82-fold higher than previously reported.