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工程化的三角褐指藻中葡萄糖-6-磷酸脱氢酶水平的升高提高了脂类积累和生长速率。

Elevated CO improves both lipid accumulation and growth rate in the glucose-6-phosphate dehydrogenase engineered Phaeodactylum tricornutum.

机构信息

Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, People's Republic of China.

Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071, People's Republic of China.

出版信息

Microb Cell Fact. 2019 Sep 23;18(1):161. doi: 10.1186/s12934-019-1214-x.

Abstract

BACKGROUND

Numerous studies have shown that stress induction and genetic engineering can effectively increase lipid accumulation, but lead to a decrease of growth in the majority of microalgae. We previously found that elevated CO concentration increased lipid productivity as well as growth in Phaeodactylum tricornutum, along with an enhancement of the oxidative pentose phosphate pathway (OPPP) activity. The purpose of this work directed toward the verification of the critical role of glucose-6-phosphate dehydrogenase (G6PDH), the rate-limiting enzyme in the OPPP, in lipid accumulation in P. tricornutum and its simultaneous rapid growth rate under high-CO (0.15%) cultivation.

RESULTS

In this study, G6PDH was identified as a target for algal strain improvement, wherein G6PDH gene was successfully overexpressed and antisense knockdown in P. tricornutum, and systematic comparisons of the photosynthesis performance, algal growth, lipid content, fatty acid profiles, NADPH production, G6PDH activity and transcriptional abundance were performed. The results showed that, due to the enhanced G6PDH activity, transcriptional abundance and NAPDH production, overexpression of G6PDH accompanied by high-CO cultivation resulted in a much higher of both lipid content and growth in P. tricornutum, while knockdown of G6PDH greatly decreased algal growth as well as lipid accumulation. In addition, the total proportions of saturated and unsaturated fatty acid, especially the polyunsaturated fatty acid eicosapentaenoic acid (EPA; C20:5, n-3), were highly increased in high-CO cultivated G6PDH overexpressed strains.

CONCLUSIONS

The successful of overexpression and antisense knockdown of G6PDH well demonstrated the positive influence of G6PDH on algal growth and lipid accumulation in P. tricornutum. The improvement of algal growth, lipid content as well as polyunsaturated fatty acids in high-CO cultivated G6PDH overexpressed P. tricornutum suggested this G6PDH overexpression-high CO cultivation pattern provides an efficient and economical route for algal strain improvement to develop algal-based biodiesel production.

摘要

背景

许多研究表明,应激诱导和基因工程可以有效地增加脂质积累,但会导致大多数微藻的生长减少。我们之前发现,升高的 CO 浓度可以提高三角褐指藻的产脂能力和生长,同时增强氧化戊糖磷酸途径(OPPP)的活性。这项工作旨在验证葡萄糖-6-磷酸脱氢酶(G6PDH)作为限速酶在三角褐指藻脂质积累中的关键作用,以及在高 CO(0.15%)培养条件下其同时快速生长的原因。

结果

在本研究中,G6PDH 被确定为藻类菌株改良的靶标,成功地在三角褐指藻中过表达和反义敲低 G6PDH,并对光合作用性能、藻类生长、脂质含量、脂肪酸谱、NADPH 产生、G6PDH 活性和转录丰度进行了系统比较。结果表明,由于 G6PDH 活性、转录丰度和 NADPH 产生的增强,过表达 G6PDH 并结合高 CO 培养导致三角褐指藻的脂质含量和生长都有显著提高,而敲低 G6PDH 则大大降低了藻类的生长和脂质积累。此外,在高 CO 培养的过表达 G6PDH 菌株中,饱和和不饱和脂肪酸的总比例,特别是多不饱和脂肪酸二十碳五烯酸(EPA;C20:5,n-3),都显著增加。

结论

G6PDH 的成功过表达和反义敲低证明了 G6PDH 对三角褐指藻生长和脂质积累的积极影响。在高 CO 培养的过表达 G6PDH 三角褐指藻中,藻类生长、脂质含量和多不饱和脂肪酸的改善表明,这种 G6PDH 过表达-高 CO 培养模式为藻类菌株改良提供了一种高效、经济的途径,以开发基于藻类的生物柴油生产。

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