Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting.
Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting.
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DOI：
10.1038/srep26327
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发表时间：
2016-05-23
影响因子：
4.6
通讯作者：
Iwata O
中科院分区：
文献类型：
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作者：
Yamada K;Suzuki H;Takeuchi T;Kazama Y;Mitra S;Abe T;Goda K;Suzuki K;Iwata O
细叶藻（Euglena gracilis）是一种单细胞鞭毛虫原生生物的微藻，近年来在大规模种植细叶藻方面取得了进展，使生产营养食品和化妆品具有成本效益，因此引起了工业界和学术界的广泛关注。此外，已知它可以产生paramylon （β-1,3-葡聚糖晶体形式）作为储备多糖，并在缺氧和厌氧条件下将其转化为蜡酯，这是生物柴油和航空生物燃料的有前途的原料。然而，在将其投入竞争激烈的燃料市场之前，仍有许多技术挑战需要解决。本文提出了一种荧光活化细胞分选（FACS）高效选育富油薄叶菊的方法。具体来说，选择育种方法是一个重复的过程，进行为期一周的异养培养，用BODIPY505/515染色细胞内脂质，并在fe离子照射野生型（WT）诱导突变后，基于facs分离高活力的富含前0.5%脂质的黄颡鱼细胞。因此，我们获得了一种活的、稳定的、富含脂质的薄叶菊突变菌株，命名为B1ZFeL，其平均脂含量比WT高40%。我们的方法为快速、经济高效、节能的生物燃料生产铺平了道路。
Euglena gracilis, a microalgal species of unicellular flagellate protists, has attracted much attention in both the industrial and academic sectors due to recent advances in the mass cultivation of E. gracilis that have enabled the cost-effective production of nutritional food and cosmetic commodities. In addition, it is known to produce paramylon (β-1,3-glucan in a crystalline form) as reserve polysaccharide and convert it to wax ester in hypoxic and anaerobic conditions–a promising feedstock for biodiesel and aviation biofuel. However, there remain a number of technical challenges to be solved before it can be deployed in the competitive fuel market. Here we present a method for efficient selective breeding of live oil-rich E. gracilis with fluorescence-activated cell sorting (FACS). Specifically, the selective breeding method is a repetitive procedure for one-week heterotrophic cultivation, staining intracellular lipids with BODIPY505/515, and FACS-based isolation of top 0.5% lipid-rich E. gracilis cells with high viability, after inducing mutation with Fe-ion irradiation to the wild type (WT). Consequently, we acquire a live, stable, lipid-rich E. gracilis mutant strain, named B1ZFeL, with 40% more lipid content on average than the WT. Our method paves the way for rapid, cost-effective, energy-efficient production of biofuel.