文献类型： 外文期刊

作者： Sun, Li-Ping ¹ ; Ouyang, Long-Ling ¹ ; Bao, Hong ¹ ; Liu, Jian-Guo ⁵ ; Sun, Zheng ¹ ; Zhou, Zhi-Gang ¹ ;

作者机构： 1.Shanghai Ocean Univ, Minist Educ, Key Lab Explorat & Utilizat Aquat Genet Resources, Shanghai 201306, Peoples R China

2.Shanghai Ocean Univ, Minist Sci & Technol, Int Res Ctr Marine Biosci, Shanghai 201306, Peoples R China

3.Shanghai Ocean Univ, Natl Demonstrat Ctr Expt Fisheries Sci Educ, Shanghai 201306, Peoples R China

4.Chinese Acad Fishery Sci, East China Sea Fisheries Res Inst, Key Lab East China Sea Fishery Resources Exploita, Minist Agr & Rural Affairs, Shanghai 200090, Peoples R China

5.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China

关键词： Myrmecia incisa; Molecular characterization; Immunoelectron microscopy; GFP fusion; Enzyme activity

期刊名称：ALGAL RESEARCH-BIOMASS BIOFUELS AND BIOPRODUCTS （ 影响因子：4.008； 五年影响因子：4.555 ）

ISSN： 2211-9264

年卷期： 2021 年 54 卷

页码：

收录情况： SCI

摘要： Microalgae represent a rich and naturally occurring source of triacylglycerol (TAG), and the microalgae-derived TAG has enormous potential in bioenergy, bio-based materials and other agricultural innovations. Glycerol-3-phosphate acyltransferase (GPAT) is a key enzyme that catalyzes the biosynthesis of TAG. Previously our lab has reported the first GPAT in the oleaginous alga Myrmecia incisa, and in the present study, a new GPAT was further identified from the same alga, which was designated as MiGPAT2. A full-length cDNA of MiGPAT2 consisting of a 1374-bp ORF, a 153-bp 5'-UTR and a 271-bp 3' -UTR was cloned. The putative protein composed of 457 amino acids, possessing the HX4D motif that acts as the activity center of GPAT catalysis. Emphasis was put on the comparison between two MiGPATs regarding subcellular localization and TAG synthesis function. A polyclonal and monoclonal antibody were separately prepared for MiGPAT1 and MiGPAT2, and the following immunogold labeling showed they were localized on chloroplast and endoplasmic reticulum (ER), respectively. This was further confirmed by the GFP fusion studies. Both MiGPATs were expressed heterologously in a GPATdeficient yeast mutant, and it was found that MiGPAT2 played key roles in the de novo TAG biosynthesis, whereas MiGPAT1 was more involved in the phospholipid formation, contributing to TAG production via an indirect way. Findings of the present study expanded our knowledge on functional and evolutionary relationships of GPAT members in M. incisa, suggesting the possibility of genetic manipulation of GPAT in this alga for enhanced TAG production.