Abstract
Genetic engineering of plants has turned out to be an attractive approach to produce various secondary metabolites. Here, we attempted to produce kynurenine, a health-promoting metabolite, in plants of Nicotiana tabacum (tobacco) transformed by Agrobacterium tumefaciens with the gene, coding for human indoleamine 2,3-dioxygenase 1 (IDO1), an enzyme responsible for the kynurenine production because of tryptophan degradation. The presence of IDO1 gene in transgenic plants was confirmed by PCR, but the protein failed to be detected. To confer higher stability to the heterologous human IDO1 protein and to provide a more sensitive method to detect the protein of interest, we cloned a gene construct coding for IDO1-GFP. Analysis of transiently transfected tobacco protoplasts demonstrated that the IDO1-GFP gene led to the expression of a detectable protein and to the production of kynurenine in the protoplast medium. Interestingly, the intracellular localisation of human IDO1 in plant cells is similar to that found in mammal cells, mainly in cytosol, but in early endosomes as well. To the best of our knowledge, this is the first report on the expression of human IDO1 enzyme capable of secreting kynurenines in plant cells.
More Information
Identification Number: | https://doi.org/10.3390/ijms22105102 |
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Status: | Published |
Refereed: | Yes |
Publisher: | MDPI |
Uncontrolled Keywords: | genetic transformation, kynurenine, protoplasts, Agrobacterium tumefaciens, Cloning, Molecular, Green Fluorescent Proteins, Humans, Indoleamine-Pyrrole 2,3,-Dioxygenase, Kynurenine, Plasmids, Protein Stability, Recombinant Proteins, Tobacco, Transformation, Bacterial, 0399 Other Chemical Sciences, 0604 Genetics, 0699 Other Biological Sciences, Chemical Physics, |
Depositing User (symplectic) | Deposited by Blomfield, Helen |
Date Deposited: | 17 Sep 2021 15:59 |
Last Modified: | 12 Jul 2024 09:23 |
Item Type: | Article |
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