We are analyzing https://link.springer.com/article/10.1186/s13068-018-1309-3.

Title:
Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis | Biotechnology for Biofuels and Bioproducts
Description:
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. 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 μg/109 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. 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.
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Keywords {🔍}

pttes, acylcoa, fatty, article, tricornutum, tag, google, scholar, acid, mutant, fig, lipid, phaeodactylum, thioesterase, cells, cas, acylcoas, knockout, min, acids, protein, increase, gene, activity, acyltransferase, accumulation, algal, plastid, cell, content, plant, increased, diatom, thioesterases, total, wildtype, tesko, coli, biosynthesis, grown, strain, nitrogen, talen, production, substrate, analysis, acyl, observed, growth, plasmid,

Topics {✒️}

primer pairs tes1up-easy-1/tes1up-easy-2 primer pair talen-tes1en-1/talen-tes1en-2 hotdog-fold thioesterase/dehydratase subfamilies long-chain acyl-coa thioesterase long-chain acyl-coa esters long-chain acyl-coa synthetases long-chain fatty acyl-coas hexane/isopropanol/water/ammonium acetate 1 medium-chain acyl-coa hydrolases pttes1-catalyzed acyl-coa hydrolysis talen-tes1en-3/talen-tes1en-4 ppha-pttes1up-shble-pttes1dn-ko19f tes1dw-easy-1/tes1dw-easy-2 helix/4-stranded sheet domains talen-based insertional mutagenesis lacks acyl-coa synthetase talen-based targeted mutagenesis ffa→acyl-coa→ffa cycle tes-ko knockout line isopropanol/water/ammonium acetate 1 isopropyl-β-d-thiogalactopyranoside hotdog-fold thioesterase localized full-length amino acids lc-ms/ms versus tlc evolutionary view thioester intermediate salicoyl-coa anr-10-labex-04 gral labex polyunsaturated acyl-coas ranging turn-α-helical ‘sausage’ [20] fatty acid-derived chemicals multi-domain enzymatic complex article download pdf fatty acid profile acyl-coa thioesterases include α/β-fold hydrolase ppha-pttes1up-shble-ko19f long-chain acyl-coenzyme hubei key laboratory genome editing technique acyl-coa profile eric maréchal author information authors 4-hbt-ii thioesterase subfamily 4-hbt-ii subfamily thioesterase fats acyl-acp thioesterases tes-ko contained significantly compartmentalized acyl-coa metabolism tal effector-nucleotide targeter fatty acid profiles acyl-coa thioesterase gene

Schema {🗺️}

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         headline:Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis
         description: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. 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 μg/109 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. 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.
         datePublished:2018-11-12T00:00:00Z
         dateModified:2018-11-12T00:00:00Z
         pageStart:1
         pageEnd:18
         license:http://creativecommons.org/publicdomain/zero/1.0/
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         keywords:
            Acyl-CoA thioesterase
            Acyl-CoA
            Fatty acids
             Phaeodactylum tricornutum
            TALEN
            Triacylglycerols
            Biotechnology
            Plant Breeding/Biotechnology
            Environmental Engineering/Biotechnology
            Renewable and Green Energy
            Microbiology
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                        name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                        type:PostalAddress
                     type:Organization
                     name:Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                     address:
                        name:Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China
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               type:Person
               name:Ling Luo
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                     name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                     address:
                        name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
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                        name:Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168, Grenoble, France
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                        name:Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168, Grenoble, France
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ScholarlyArticle:
      headline:Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis
      description: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. 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 μg/109 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. 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.
      datePublished:2018-11-12T00:00:00Z
      dateModified:2018-11-12T00:00:00Z
      pageStart:1
      pageEnd:18
      license:http://creativecommons.org/publicdomain/zero/1.0/
      sameAs:https://doi.org/10.1186/s13068-018-1309-3
      keywords:
         Acyl-CoA thioesterase
         Acyl-CoA
         Fatty acids
          Phaeodactylum tricornutum
         TALEN
         Triacylglycerols
         Biotechnology
         Plant Breeding/Biotechnology
         Environmental Engineering/Biotechnology
         Renewable and Green Energy
         Microbiology
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      isPartOf:
         name:Biotechnology for Biofuels
         issn:
            1754-6834
         volumeNumber:11
         type:
            Periodical
            PublicationVolume
      publisher:
         name:BioMed Central
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:Xiahui Hao
            affiliation:
                  name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                     type:PostalAddress
                  type:Organization
                  name:Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Ling Luo
            affiliation:
                  name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                     type:PostalAddress
                  type:Organization
                  name:Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Juliette Jouhet
            affiliation:
                  name:Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168
                  address:
                     name:Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168, Grenoble, France
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                     name:Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168, Grenoble, France
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            name:Eric Maréchal
            affiliation:
                  name:Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168
                  address:
                     name:Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, Institut National de la Recherche Agronomique, Université Grenoble Alpes, UMR 5168, Grenoble, France
                     type:PostalAddress
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            type:Person
            name:Hanhua Hu
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                  name:Institute of Hydrobiology, Chinese Academy of Sciences
                  address:
                     name:Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
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                  type:Organization
            type:Person
            name:Yufang Pan
            affiliation:
                  name:Institute of Hydrobiology, Chinese Academy of Sciences
                  address:
                     name:Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Xiaoming Tan
            affiliation:
                  name:Hubei University
                  address:
                     name:Hubei Collaborative Innovation Center for Green Transformation of Bioresources, Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei University, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Zhuo Chen
            affiliation:
                  name:Shandong Normal University
                  address:
                     name:Shandong Provincial Key Laboratory of Plant Stress, College of Life Science, Shandong Normal University, Jinan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Lingjie You
            affiliation:
                  name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                     type:PostalAddress
                  type:Organization
                  name:Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Hong Chen
            affiliation:
                  name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                     type:PostalAddress
                  type:Organization
                  name:Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Fang Wei
            affiliation:
                  name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                     type:PostalAddress
                  type:Organization
                  name:Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Hubei Key Laboratory of Lipid Chemistry and Nutrition, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Yangmin Gong
            affiliation:
                  name:Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences
                  address:
                     name:Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, People’s Republic of China
                     type:PostalAddress
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