Human indoleamine 2,3-dioxygenase 1 (Ido1) expressed in plant cells induces kynurenine production

TY - JOUR

T1 - Human indoleamine 2,3-dioxygenase 1 (Ido1) expressed in plant cells induces kynurenine production

AU - Bellucci, Michele

AU - Pompa, Andrea

AU - De Marcos Lousa, Carine

AU - Panfili, Eleonora

AU - Orecchini, Elena

AU - Maricchiolo, Elisa

AU - Fraternale, Daniele

AU - Orabona, Ciriana

AU - De Marchis, Francesca

AU - Pallotta, Maria Teresa

N1 - Funding Information: This research was funded by Ministero dell’Istruzione, dell’Università e della Ricerca (MIUR), grant number PRIN 2017WJZ9W9 to M.T.P. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2021/5/12

Y1 - 2021/5/12

N2 - 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.

AB - 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.

KW - Genetic transformation

KW - Kynurenine

KW - Protoplasts

UR - https://www.scopus.com/pages/publications/85105446624

U2 - 10.3390/ijms22105102

DO - 10.3390/ijms22105102

M3 - Article

C2 - 34065885

AN - SCOPUS:85105446624

SN - 1661-6596

VL - 22

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

IS - 10

M1 - 5102

ER -