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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; Chen, Hong1,2; Wei, Fang1,2; Gong, Yangmin1,2
Corresponding AuthorGong, Yangmin(gongyangmin@caas.cn)
2018-11-12
Source PublicationBIOTECHNOLOGY FOR BIOFUELS
ISSN1754-6834
Volume11Pages:18
AbstractBackground: 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.
KeywordAcyl-CoA thioesterase Acyl-CoA Fatty acids Phaeodactylum tricornutum TALEN Triacylglycerols
DOI10.1186/s13068-018-1309-3
Funding OrganizationNational 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps)
Indexed BySCI ; SCI
Language英语
Funding ProjectNational 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 Research AreaBiotechnology & Applied Microbiology ; Energy & Fuels
WOS SubjectBiotechnology & Applied Microbiology ; Energy & Fuels
WOS IDWOS:000451082100001
WOS KeywordCARRIER PROTEIN THIOESTERASE ; ACYL-COA THIOESTERASE ; MOLECULAR CHARACTERIZATION ; LIPID-SYNTHESIS ; FATTY-ACIDS ; CHAIN ; ACYLTRANSFERASE ; BIOSYNTHESIS ; ACCUMULATION ; TRANSFORMATION
PublisherBMC
Funding OrganizationNational 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; 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 ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (Institut Carnot 3BCAR) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps) ; Agence Nationale de la Recherche (IDEX UGA CDP Glyco@Alps)
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Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/28983
Collection藻类生物学及应用研究中心_期刊论文
Corresponding AuthorGong, Yangmin
Affiliation1.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
Recommended Citation
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,2018,11:18.
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,11,18.
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(2018):18.
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