半连续稀释异养-贫营养两段法培育富油小球藻及调控机制

项目名称： 半连续稀释异养-贫营养两段法培育富油小球藻及调控机制

项目编号： No.21306161

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 化学工业

项目作者： 李玉芹

作者单位： 湘潭大学

项目金额： 25万元

中文摘要： 如何提高微藻细胞生物量并有效富集胞内油脂一直是微藻能源领域的一大难题，也是微藻生物柴油实现产业化的关键。我们前期研究结果表明，两段培养法可以促使小球藻（Chlorella protothecoides）生物量的提高及胞内油脂的大量积累，这种现象产生的原因尚不明确。本项目采用半连续稀释异养-贫营养两段法模式培育小球藻，拟以异养高密度藻细胞和缺氮积脂细胞为对照组，两段法培育积脂藻细胞为实验组，应用蛋白组学、GO富集、KEGG代谢通路、活体共聚焦显微（CLSM）、流式细胞术（FCM）等现代生物学集成研究手段，全面系统地分析不同培养条件、不同培养阶段小球藻的生理生化变化及其蛋白质组的表达差异，甄选与小球藻油脂富集有关的代谢调控因子（即油脂合成积累途径的调控酶），揭示调控因子响应两段培育富集油脂的调控机制，为人工干预微藻油脂积累、推进微藻生物柴油规模化发展提供基础，同时也为其它藻类代谢研究提供参考。

中文关键词： 半连续稀释异养-贫营养；小球藻；油脂；调控酶；调控机制

英文摘要： The improvement of microalgal biomass and intracellular lipid accumulation is a big challenge in the field of microalgal energy, which correlates directly with the commercial development of microalgal biodiesel. A two-stage cultivation model has been proposed to promote the increase in Chlorella protothecoides biomass and accumulation of intracellular lipid by our preliminary results. However, the fundamental mechanism underlying this promotion effect is not clear yet. In this proposal, the semi-continuous dilution heterotrophic-N deprivation two-step cultivation model will be employed to culture rich-oil Chlorella protothecoides. And the heterotrophic high-density Chlorella cells and nitrogen deprived rich-oil Chlorella cells will be adopted as control group, the two-stage cultured rich-oil Chlorella cells will be adopted as the experimental group to carry out study. The integrated modern biological means based on proteomics, Gene Ontology,KEGG (Kyoto encyclopedia of genes and genomes) metabolic pathway, in vivo confocal laser scanning microscopy (CLSM) and flow cytometry (FCM) will be employed to comprehensively and systematically analysize the physiological and biochemical changes and differentially expressed proteins in Chlorella cells. The aims of this proposal is to screen metabolic regulatory factors rela

英文关键词： Semi-continuous dilution heterotrophy-N starvation；Chlorella protothecoides；Lipids；Regulated enzymes；Regulated mechanism