The role of microtubules in microalgae: promotion of lipid accumulation and extraction.

Lijie Zhang, Xiao Lin, Zhigang Yang, Liqun Jiang, Qingjie Hou, Zhen Xie, Yizhen Li, Haiyan Pei
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Abstract

Background: Microtubules in cells are closely related to the growth and metabolism of microalgae. To date, the study of microalgal microtubules has mainly concentrated on revealing the relationship between microtubule depolymerization and synthesis of precursors for flagellar regeneration. While information on the link between microtubule depolymerization and biosynthesis of precursors for complex organic matter (such as lipid, carbohydrate and protein), is still lacking, a better understanding of this could help to achieve a breakthrough in lipid regulation. With the aim of testing the assumption that microtubule disruption could regulate carbon precursors and redirect carbon flow to promote lipid accumulation, Chlorella sorokiniana SDEC-18 was pretreated with different concentrations of oryzalin.

Results: Strikingly, microalgae that were pretreated with 1.5 mM oryzalin accumulated lipid contents of 41.06%, which was attributed to carbon redistribution induced by microtubule destruction. To promote the growth of microalgae, two-stage cultivation involving microtubule destruction was employed, which resulted in the lipid productivity being 1.44 times higher than that for microalgae with routine single-stage cultivation, as well as yielding a desirable biodiesel quality following from increases in monounsaturated fatty acid (MUFA) content. Furthermore, full extraction of lipid was achieved after only a single extraction step, because microtubule destruction caused removal of cellulose synthase and thereby blocked cellulose biosynthesis.

Conclusions: This study provides an important advance towards observation of microtubules in microalgae through immunocolloidal gold techniques combined with TEM. Moreover, the observation of efficient lipid accumulation and increased cell fragility engendered by microtubule destruction has expanded our knowledge of metabolic regulation by microtubules. Finally, two-stage cultivation involving microtubule destruction has established ideal growth, coupling enhanced lipid accumulation and efficient oil extraction; thus gaining advances in both applied and fundamental research in algal biodiesel production.

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微管在微藻中的作用:促进脂质积累和提取。
背景:细胞中的微管与微藻的生长和代谢密切相关。迄今为止,对微藻微管的研究主要集中在揭示微管解聚与鞭毛再生前体合成之间的关系。虽然关于微管解聚与复杂有机物(如脂质、碳水化合物和蛋白质)前体生物合成之间的联系的信息仍然缺乏,但更好地了解这一点有助于在脂质调节方面取得突破。为了验证微管破坏可以调节碳前体并改变碳流动方向以促进脂质积累的假设,我们用不同浓度的米草酸苷预处理小球藻stec -18。结果:显著的是,经1.5 mM米扎林预处理的微藻脂质含量为41.06%,这是由于微管破坏引起的碳重分布。为了促进微藻的生长,采用了涉及微管破坏的两阶段培养,其脂质产量比常规单阶段培养的微藻高1.44倍,并且由于单不饱和脂肪酸(MUFA)含量的增加,产生了理想的生物柴油质量。此外,由于微管破坏导致纤维素合酶的去除,从而阻碍了纤维素的生物合成,因此只需一个提取步骤就可以完全提取脂质。结论:本研究为利用免疫胶体金技术结合透射电镜观察微藻微管提供了重要进展。此外,微管破坏引起的高效脂质积累和细胞脆弱性增加的观察扩大了我们对微管代谢调节的认识。最后,微管破坏的两阶段培养建立了理想的生长,耦合了脂质积累和高效的油脂提取;因此,在藻类生物柴油生产的应用和基础研究方面取得了进展。
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