Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis | |
Hao, Xiahui1,2; Luo, Ling1,2; Jouhet, Juliette3; Rebeille, Fabrice3; Marechal, Eric3; Hu, Hanhua4; Pan, Yufang4; Tan, Xiaoming5; Chen, Zhuo6; You, Lingjie1,2 | |
刊名 | BIOTECHNOLOGY FOR BIOFUELS |
2018-11-12 | |
卷号 | 11期号:1页码:18 |
关键词 | Acyl-CoA thioesterase Acyl-CoA Fatty acids Phaeodactylum tricornutum TALEN Triacylglycerols |
ISSN号 | 1754-6834 |
DOI | 10.1186/s13068-018-1309-3 |
英文摘要 | Background: In photosynthetic oleaginous microalgae, acyl-CoA molecules are used as substrates for the biosynthesis of membrane glycerolipids, triacylglycerol (TAG) and other acylated molecules. Acyl-CoA can also be directed to beta-oxidative catabolism. They can be utilized by a number of lipid metabolic enzymes including endogenous thioesterases, which catalyze their hydrolysis to release free fatty acids. Acyl-CoA availability thus plays fundamental roles in determining the quantity and composition of membrane lipids and storage lipids. Results: Here, we have engineered the model diatom Phaeodactylum tricornutum to produce significantly increased TAGs by disruption of the gene encoding a Hotdog-fold thioesterase involved in acyl-CoA hydrolysis (ptTES1). This plastidial thioesterase can hydrolyze both medium-and long-chain fatty acyl-CoAs, but has the highest activity toward long-chain saturated and monounsaturated fatty acyl-CoAs. The maximum rate was found with oleoyl-CoA, which is hydrolyzed at 50 nmol/min/mg protein. The stable and targeted interruption of acyl-CoA thioesterase gene was achieved using a genome editing technique, transcription activator-like effector nucleases (TALENs). Disruption of native ptTES1 gene resulted in a 1.7-fold increase in TAG content when algal strains were grown in nitrogen-replete media for 8 days, whereas the content of other lipid classes, including phosphoglycerolipids and galactoglycerolipids, remained almost unchanged. The engineered algal strain also exhibited a marked change in fatty acid profile, including a remarkable increase in 16: 0 and 16: 1 and a decrease in 20:5. Nitrogen deprivation for 72 h further increased TAG content and titer of the engineered strain, reaching 478 mu g/10(9) cells and 4.8 mg/L, respectively. Quantitative determination of in vivo acyl-CoAs showed that the total acyl-CoA pool size was significantly higher in the engineered algal strain than that in the wild type. Conclusions: This study supports the role of ptTES1 in free fatty acid homeostasis in the plastid of Phaeodactylum and demonstrates the potential of TALEN-based genome editing technique to generate an enhanced lipid-producing algal strain through blocking acyl-CoA catabolism. |
资助项目 | National Natural Science Foundation of China[31100072] ; National Natural Science Foundation of China[41776175] ; National Natural Science Foundation of China[31600286] ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences[CAAS-ASTIP-2013-OCRI] ; Agence Nationale de la Recherche[ANR-10-LABEX-04] ; Agence Nationale de la Recherche[ANR-11-BTBR-0008] ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) |
WOS关键词 | CARRIER PROTEIN THIOESTERASE ; ACYL-COA THIOESTERASE ; MOLECULAR CHARACTERIZATION ; LIPID-SYNTHESIS ; FATTY-ACIDS ; CHAIN ; ACYLTRANSFERASE ; BIOSYNTHESIS ; ACCUMULATION ; TRANSFORMATION |
WOS研究方向 | Biotechnology & Applied Microbiology ; Energy & Fuels |
语种 | 英语 |
出版者 | BMC |
WOS记录号 | WOS:000451082100001 |
资助机构 | National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; Agricultural Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences ; 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内容类型 | 期刊论文 |
源URL | [http://ir.ihb.ac.cn/handle/342005/28982] |
专题 | 水生生物研究所_藻类生物学及应用研究中心_期刊论文 |
通讯作者 | Gong, Yangmin |
作者单位 | 1.Chinese Acad Agr Sci, Oil Crops Res Inst, Minist Agr, Key Lab Biol & Genet Improvement Oil Crops, 2 Xudong Second Rd, Wuhan 430062, Peoples R China 2.Chinese Acad Agr Sci, Oil Crops Res Inst, Hubei Key Lab Lipid Chem & Nutr, Wuhan 430062, Peoples R China 3.Univ Grenoble Alpes, INRA, Commissariat Energie Atom & Energies Alternat, Lab Physiol Cellulaire & Vegetale,CNRS,UMR 5168, F-38041 Grenoble, France 4.Chinese Acad Sci, Inst Hydrobiol, Key Lab Algal Biol, Wuhan 430072, Peoples R China 5.Hubei Univ, Coll Life Sci, Hubei Key Lab Ind Biotechnol, Hubei Collaborat Innovat Ctr Green Transformat Bi, Wuhan 430062, Peoples R China 6.Shandong Normal Univ, Coll Life Sci, Shandong Prov Key Lab Plant Stress, Jinan 250014, Shandong, Peoples R China |
推荐引用方式 GB/T 7714 | Hao, Xiahui,Luo, Ling,Jouhet, Juliette,et al. Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis[J]. BIOTECHNOLOGY FOR BIOFUELS,2018,11(1):18. |
APA | Hao, Xiahui.,Luo, Ling.,Jouhet, Juliette.,Rebeille, Fabrice.,Marechal, Eric.,...&Gong, Yangmin.(2018).Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis.BIOTECHNOLOGY FOR BIOFUELS,11(1),18. |
MLA | Hao, Xiahui,et al."Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis".BIOTECHNOLOGY FOR BIOFUELS 11.1(2018):18. |
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