School of Biological Engineering, Guizhou Medical University, Guiyang, Guizhou Province, China.
Departments of Pharmacology, Biochemistry and Biophysics, and Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
PLoS One. 2020 Mar 20;15(3):e0229556. doi: 10.1371/journal.pone.0229556. eCollection 2020.
The heterotrophic microalgae Crypthecodinium cohnii was usually cultivated in complex medium containing glucose, yeast extract and sea salt. For the preparation of DHA with highest purity, a new defined medium without the yeast extract was developed. Different inoculated densities, C/N ratios, temperatures, culture volumes and glucose additions were investigated to optimize the algal growth rate and DHA production. The growth period in C. cohnii was shortened from 12-14 days to 7-8 days, the OD600 was enhanced from 2.0 to 3.0, the glucose consumption was accelerated and used up on day 3-4, and the DHA content in culture were increased from 10 to 45 nmoles/300 μl batch. It was found that C. cohnii had optimal growth and DHA accumulation in 25 °C, 0.2 inoculated density, 5-10 C/N ratio, 5:1 air/culture volume ratio. This is the first time DHA production using C.cohnii has been optimized in synthetic medium. This allows preparation of uniformly radiolabeled 13C- and 14C-DHA.
通常情况下,异养微藻裂殖壶菌被培养在含有葡萄糖、酵母提取物和海盐的复杂培养基中。为了制备最高纯度的 DHA,开发了一种不含酵母提取物的新型限定培养基。研究了不同的接种密度、C/N 比、温度、培养体积和葡萄糖添加量,以优化藻类的生长速度和 DHA 产量。裂殖壶菌的生长周期从 12-14 天缩短到 7-8 天,OD600 从 2.0 提高到 3.0,葡萄糖消耗在第 3-4 天加速并耗尽,培养物中的 DHA 含量从 10 增加到 45nmoles/300μl 批次。研究发现,在 25°C、0.2 的接种密度、5-10 的 C/N 比、5:1 的空气/培养物体积比下,裂殖壶菌的生长和 DHA 积累达到最佳状态。这是首次在合成培养基中对裂殖壶菌的 DHA 生产进行优化。这使得均一放射性标记的 13C-和 14C-DHA 的制备成为可能。