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Cloning and characterization of a novel diacylglycerol acyltransferase from the diatom Phaeodactylum tricornutum

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Abstract

In this study, a cDNA encoding a novel acyl-CoA:diacylglycerol acyltransferase (DGAT)-like protein is identified and isolated from the diatom microalga Phaeodactylum tricornutum (PtDGAT3). Analysis of the sequence reveals that ptDGAT3 cDNA encodes a protein of 504 amino acids with a molecular mass of 64.5 KDa. The putative ptDGAT3 protein has two catalytic domains: a wax ester synthase-like acyl-CoA acyltransferase domain and a bacteria-specific acyltransferase domain, which shows higher similarity to the DGAT3 of Acinetobacter calcoaceticus than reported DGAT1 or DGAT2 from high plants or algae. Its activity was confirmed by heterologous expression of PtDGAT3 in a neutral lipid-deficient quadruple mutant yeast Saccharomyces cerevisiae H1246. The recombinant yeast restored the formation of a lipid body and displayed a preference to the incorporation of unsaturated C18 fatty acids into triacyglycerol (TAG). This is the first characterized algal DGAT3 gene, giving further evidence to the occurrence of a DGAT3-mediated TAG biosynthesis pathway.

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Acknowledgments

This work was supported by grants from National Key Basic Research Program of China (2011CB200902), Ocean Public Welfare Scientific Research Project, State Oceanic Administration of China (200905021-3, 201205027), Nature Science Funds for Distinguished Young Scholar of Shandong Province (JQ200914), National Natural Science Foundation of China (41176144), Natural Science Foundation of Jiangsu Province (BK2012650).

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Authors and Affiliations

  1. Key Laboratory of Experimental Marine Biology, Experimental Marine Biology Institute of Oceanology, Chinese Academy of Sciences, No. 7 Nanhai Road, Shinan District, Qingdao, 266071, Shandong Province, China

    Guoting Zheng & Peng Jiang

  2. Key Laboratory of Coastal Biology and Biological Resources Utilization, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, No. 17 Chunhui Road, Laishan District, Yantai, 264003, Shandong Province, China

    Yulin Cui, Xueqin Li & Song Qin

  3. Key Laboratory of Marine Biotechnology, Ningbo University, Ningbo, 315211, China

    Guoting Zheng

  4. College of Life Sciences, Shenzhen University, Shenzhen, 518060, China

    Xueqin Li

  5. Institute of Marine and Coastal Sciences, Rutgers University, New Brunswick, 08901, NJ, USA

    Hanzhi Lin

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  1. Yulin Cui
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  2. Guoting Zheng
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  3. Xueqin Li
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  4. Hanzhi Lin
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  5. Peng Jiang
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  6. Song Qin
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Corresponding authors

Correspondence to Peng Jiang or Song Qin.

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Yulin Cui and Guoting Zheng contributed equally to the work.

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Cui, Y., Zheng, G., Li, X. et al. Cloning and characterization of a novel diacylglycerol acyltransferase from the diatom Phaeodactylum tricornutum . J Appl Phycol 25, 1509–1512 (2013). https://doi.org/10.1007/s10811-013-9991-9

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  • DOI: https://doi.org/10.1007/s10811-013-9991-9

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