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On October 9, The Plant Journal, an authoritative journal in the field of botany, published the results of Professor Li Maoteng's research group at the School of Life Science and Technology of Huazhong University of Science and Technology, with the paper "Class A lysophosphatidic acid acyltransferase 2 from Camelina sativa promotes very long-chain fatty acids.
" accumulation in phospholipid and triacylglycerol" (paper link: https://onlinelibrary.
wiley.
com/doi/10.
1111/tpj.
15999).
This study is the first to discover the key genes that promote the accumulation of ultra-long-chain fatty acids on phospholipids, providing a breakthrough for breaking through the technical bottleneck of ultra-long-chain fatty acid production
.
Ultra-long chain fatty acids (VLCFAs) are a class of fatty acids with a carbon chain length greater than 18 carbons, which are of great value
in industrial production and food and health care.
Taking erucic acid (C22:1) as an example, erucic acid molecule carbon chain long, hydrophobicity and waterproofness, excellent lubrication performance, is an important oleochemical product, in metallurgy, machinery, rubber, chemical industry, paint, textile, manufacturing and medicine and other fields have a wide range of uses
.
At present, the source of VLCFAs for industrial use is mainly extracted from plant seeds and deep-sea fish in nature, which is far from meeting the needs of
industrial use.
The seeds of some varieties (lines) or germplasm resources of cruciferous plants, such as sea cabbage and rape, are rich in VLCFAs-erucic acid, which can reach more than
50%.
Therefore, further increasing the content of VLCFAs in cruciferous plants has been one of
the hot spots of current research.
In recent years, genetic engineering to increase VLCFAs content in oil crop seeds has achieved some success, but it has also encountered technical bottlenecks
.
For example, the oil content of genetically modified seeds decreased, the content of VLCFAs on phosphatidylcholine (PC) was low, and the accumulation of VLCFAs in triglycerides (TAG) could
not be further improved.
Therefore, it is of great value
to find genes that promote the accumulation of VLCFAs in PC and apply them in genetic engineering and synthetic biology.
The team performed a functional analysis of the lysophosphatidyl acid acyltransferase (LPAT) family at the genome-wide level of water chestnut and found that the overexpression of LPAT2 belonging to class A in Arabidopsis seeds significantly increased the content of VLCFAs, and the oil content was also increased
at a certain level.
In addition, the group also found that the overexpression of LPAT2 from representative cruciferous plants such as Arabidopsis, rape, cabbage and kale can also promote the accumulation of VLCFAs in Arabidopsis seeds, proving that the gene may have functional conservancy
.
Based on this, it is speculated that LPAT2 may have VLCFAs substrate preference
.
However, the results of in vitro enzyme activity and substrate preference experiments showed that although the overexpression line of LPAT2 increased the activity of VLCFAs, the carbon level analysis results of TAG molecules also confirmed that LPAT2 could slightly promote the accumulation
of VLCFAs at the sn-2 position.
Figure 1 The LPAT2 superexpression line and control seeds contain VLCFAs of PA Molecular percentage content
In order to further reveal the mechanism by which water chestnut LPAT2 promotes VLCFAs accumulation, the molecular composition of lipid intermediate metabolites was detected
by ESI-MS/MS.
It was found that the proportion of phospholipid molecules containing VLCFAs in the superexpression line seeds was significantly increased (Figure 1).
Further analysis showed that the percentage content of VLCFAs in superexpression seeds was more than 4 times higher than that in the control group, which was more than 4 times higher than that of the control group, in the direct product of LPAT2.
The percentage content of VLCFAs in phosphatidylcholine (PC) and phosphatidylethanolamine (PE), the main components of phospholipids, increased by 2-2.
8 times; This is consistent with the mass spectrometry test results (Figure 2).
Prior to this, no LPAT overexpression that promotes the accumulation of VLCFAs in phospholipids has been reported
.
More importantly, the proportion of double C20 molecules in phospholipid molecules has increased significantly, which provides a breakthrough for breaking through the technical bottleneck of further accumulation of VLCFAs in
seeds.
Figure 2 Relative content of VLCFAs in the Aqualid LPAT2 superexpression line and control seeds in lipid intermediate metabolites such as phospholipids
The corresponding author of this paper is Professor Li Maoteng of the School of Life Science and Technology, Dr.
Yin Yongtai and Dr.
Nadia Raboanatahiry are the co-first authors of the paper, doctoral student Chen Kang, master student Guo Zhenyi and other students and members of the innovation and entrepreneurship training program project team from Huazhong University of Science and Technology also participated in the relevant research work
.
This research was supported
by the National Natural Science Foundation of China (32172087), the China Postdoctoral Fund (2016M602309) and other projects.