Antisense knockdown of pyruvate dehydrogenase kinase promotes the neutral lipid accumulation in the diatom Phaeodactylum tricornutum
| Autor: | Zhi-Kai Yang, Ying-Fang Niu, Zhong-Ming Wang, Xiang Wang, Jie-Sheng Liu, Wei-Dong Yang, Meng-Han Zhang, Yu-Han Ma, Hong-Ye Li |
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| Rok vydání: | 2014 |
| Předmět: |
Pyruvate decarboxylation
Pyruvate dehydrogenase kinase Bioengineering PKM2 Pyruvate dehydrogenase phosphatase Protein Serine-Threonine Kinases Applied Microbiology and Biotechnology Biofuel Microalgae RNA Antisense Phaeodactylum tricornutum Antisense Diatoms biology Research fungi Pyruvate Dehydrogenase Acetyl-Transferring Kinase Lipid Pyruvate dehydrogenase complex biology.organism_classification Lipid Metabolism Pyruvate carboxylase Biochemistry Metabolic Engineering Gene Knockdown Techniques Branched-chain alpha-keto acid dehydrogenase complex Biotechnology Microalga |
| Zdroj: | Microbial Cell Factories |
| ISSN: | 1475-2859 |
| Popis: | Background Microalgae have been an emerging biofuel resource; however, the germplasm improvement has been slow due to the lack of molecular tools. Pyruvate dehydrogenase kinase (PDK) deactivates the pyruvate dehydrogenase complex (PDC) which catalyzes the oxidative decarboxylation of pyruvate. Acetyl-CoA production via PDC is important in plant tissues that are active in fatty acid synthesis. Results A 1261-bp cDNA of a putative PDK gene (PtPDK) was cloned from a diatom Phaeodactylum tricornutum, and PtPDK antisense knockdown transgenic diatoms were generated. Both PtPDK transcript abundance and enzyme activity were reduced significantly due to antisense knockdown of PtPDK. Neutral lipid content of transgenic diatom cells increased up to 82% as determined by Nile red staining, and fatty acid composition was not altered. Transgenic cells showed slightly lower growth rate but similar cell size with the wild type, hence retaining similar biomass productivity. Conclusions This work first obtained a successful engineered diatom regulating a key gene involved in lipid metabolism. Our findings also provide powerful indications in enhancing microalgal lipid production by metabolic engineering for biofuel industry. |
| Databáze: | OpenAIRE |
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