Improvement of Neutral Lipid and Polyunsaturated Fatty Acid Biosynthesis by Overexpressing a Type 2 Diacylglycerol Acyltransferase in Marine Diatom Phaeodactylum tricornutum
Abstract
:1. Introduction
2. Results
2.1. Sequence-Structure Analysis of DGAT2 Protein
2.2. Photosynthesis Activity and Growth of Transgenic P. tricornutum
2.3. Analysis on Lipid Content and Fatty Acid Composition
Fatty Acid | Wild Type | DGAT2 |
---|---|---|
C14:0 | 1.62 | 1.36 |
C15:0 | 0.18 | 0.36 |
C16:0 | 6.43 | 6.69 |
C17:0 | 0.1 | 0.05 |
C18:0 | 3.35 | 5.5 |
C20:0 | 0.08 | 1.5 |
C21:0 | 0.49 | 0.48 |
C22:0 | 0.05 | 0.15 |
C24:0 | 0.7 | 1.54 |
SUM SFA | 13 | 17.63 |
C16:1 | 6.11 | 9.31 |
C18:1 | 1.03 | 0.29 |
C22:1 | ND | 0.04 |
C24:1 | ND | 0.04 |
SUM MUFA | 7.14 | 9.68 |
C16:3 | 2.26 | 1.87 |
C18:2 | 0.78 | 0.61 |
C18:3 | 0.08 | ND |
C20:2 | 0.05 | 0.05 |
C20:5 | 4.21 | 7.42 |
C22:6 | ND | ND |
SUM PUFA | 7.38 | 9.95 |
2.4. Expression of DGAT2 Determined by qPCR and Western Blot Analysis
3. Discussion
4. Experimental Section
4.1. Diatom Strain and Culture Conditions
4.2. Cloning of DGAT2 and Generation of Transgenic P. tricornutum by Electroporation
4.3. Selection and PCR Screening of Transgenic Microalgae
4.4. Measurement of Photosynthesis Activity
4.5. Lipid Analysis
4.6. Transcript Abundance of DGAT2 Analyzed by qPCR
4.7. DGAT2 Protein Expression Analyzed by Western Blot Analysis
5. Conclusions
Abbreviations
CAT | chloramphenicol acetyltransferase |
DGAT | diacylglycerol acyltranferase |
Fcp | fucoxanthin chlorophyll a/c binding protein |
NCBI | National Center for Biotechnology Information |
PAGE | polyacrylamide gel electrophoresis |
PUFA | polyunsaturated fatty acid |
TAG | triacylglyceride |
Acknowledgements
Conflicts of Interest
References
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Niu, Y.-F.; Zhang, M.-H.; Li, D.-W.; Yang, W.-D.; Liu, J.-S.; Bai, W.-B.; Li, H.-Y. Improvement of Neutral Lipid and Polyunsaturated Fatty Acid Biosynthesis by Overexpressing a Type 2 Diacylglycerol Acyltransferase in Marine Diatom Phaeodactylum tricornutum. Mar. Drugs 2013, 11, 4558-4569. https://doi.org/10.3390/md11114558
Niu Y-F, Zhang M-H, Li D-W, Yang W-D, Liu J-S, Bai W-B, Li H-Y. Improvement of Neutral Lipid and Polyunsaturated Fatty Acid Biosynthesis by Overexpressing a Type 2 Diacylglycerol Acyltransferase in Marine Diatom Phaeodactylum tricornutum. Marine Drugs. 2013; 11(11):4558-4569. https://doi.org/10.3390/md11114558
Chicago/Turabian StyleNiu, Ying-Fang, Meng-Han Zhang, Da-Wei Li, Wei-Dong Yang, Jie-Sheng Liu, Wei-Bin Bai, and Hong-Ye Li. 2013. "Improvement of Neutral Lipid and Polyunsaturated Fatty Acid Biosynthesis by Overexpressing a Type 2 Diacylglycerol Acyltransferase in Marine Diatom Phaeodactylum tricornutum" Marine Drugs 11, no. 11: 4558-4569. https://doi.org/10.3390/md11114558