文献类型：期刊文献

英文题名：Dual expression of plastidial GPAT1 and LPAT1 regulates triacylglycerol production and the fatty acid profile in Phaeodactylum tricornutum

作者：Wang, Xiang[1];Dong, Hong-Po[2,3];Wei, Wei[1];Balamurugan, Srinivasan[1];Yang, Wei-Dong[1];Liu, Jie-Sheng[1];Li, Hong-Ye[1]

机构：[1]Jinan Univ, Key Lab Eutrophicat & Red Tide Prevent, Guangdong Higher Educ Inst, Coll Life Sci, Guangzhou 510632, Guangdong, Peoples R China;[2]Guangdong Ocean Univ, Sch Ocean & Meteorol, Zhanjiang 524088, Peoples R China;[3]East China Normal Univ, State Key Lab Estuarine & Coastal Res, Shanghai 200062, Peoples R China

年份：2018

卷号：11

外文期刊名：BIOTECHNOLOGY FOR BIOFUELS

收录：SCI-EXPANDED(收录号：WOS:000450991800002)、、WOS

基金：This study was funded by the National Natural Science Foundation of China (41725002, 41671463, 41576123, 41576132), the Guangdong Natural Science Foundation (2018A030313164, 2016A030312004, 2014A030308010), and the innovation and strengthening project of Guangdong Ocean University (GDOU2014050201, GDOU2013010203, GDOU2013050201). We thank Prof. Han-Jia Lin from National Taiwan Ocean University for kindly providing the vector pPhAP1 for protein subcellular localization.

语种：英文

外文关键词：Diatoms; GPAT1; LPAT1; Biofuel; Triacylglycerol biosynthesis; Prokaryotic TAG pathway

外文摘要：BackgroundMetabolic engineering has emerged as a potential strategy for improving microalgal lipid content through targeted changes to lipid metabolic networks. However, the intricate nature of lipogenesis has impeded metabolic engineering. Therefore, it is very important to identify the crucial metabolic nodes and develop strategies to exploit multiple genes for transgenesis. In an attempt to unravel the microalgal triacylglycerol (TAG) pathway, we overexpressed two key lipogenic genes, glycerol-3-phosphate acyltransferase (GPAT1) and lysophosphatidic acid acyltransferase (LPAT1), in oleaginous Phaeodactylum tricornutum and determined their roles in microalgal lipogenesis.ResultsEngineered P. tricornutum strains showed enhanced growth and photosynthetic efficiency compared with that of the wild-type during the growth phase of the cultivation period. However, both the cell types reached stationary phase on day 7. Overexpression of GPAT1 and LPAT1 increased the TAG content by 2.3-fold under nitrogen-replete conditions without compromising cell growth, and they also orchestrated the expression of other key genes involved in TAG synthesis. The transgenic expression of GPAT1 and LPAT1 influenced the expression of malic enzyme and glucose 6-phosphate dehydrogenase, which enhanced the levels of lipogenic NADPH in the transgenic lines. In addition, GPAT1 and LPAT1 preferred C16 over C18 at the sn-2 position of the glycerol backbone.ConclusionOverexpression of GPAT1 together with LPAT1 significantly enhanced lipid content without affecting growth and photosynthetic efficiency, and they orchestrated the expression of other key photosynthetic and lipogenic genes. The lipid profile for elevated fatty acid content (C16-CoA) demonstrated the involvement of the prokaryotic TAG pathway in marine diatoms. The results suggested that engineering dual metabolic nodes should be possible in microalgal lipid metabolism. This study also provides the first demonstration of the role of the prokaryotic TAG biosynthetic pathway in lipid overproduction and indicates that the fatty acid profile can be tailored to improve lipid production.