Assessment of Solanum tuberosum and Phytophthora infestans dihydroorotate dehydrogenase enzyme as an alternative target to control late blight

Abstract

"In this study we aimed to characterize the dihydroorotase dehydrogenase (DHOD), which catalyzes the fourth and only redox step of the pathway, and to reveal the differences between the enzymes from the pathogen P. infestans (PiDHOD) and the host, Solanum tuberosum (StDHOD). A full length and truncated StDHODs were overexpressed in Eschericlua coli but could not be purified and characterized because they were insoluble. A truncated P. Infestans recombinant protein, AN-PiDHOD was purified and the kinetic parameters were calculated using the Michaelis-Menten model to obtain a Km of 14.7 PM for dihydroorotate, Km of 6 HM for Qd and a v max of 5.85 gmol min-lmg-l at 30 0C. We also investigated the importance of the transmembrane signal peptide in the mitochondrial localization of PiDHOD. We constructed Six different C-terminal fusions of the PiDHOD with GFP to examine subcellular localization of the protein in transgenic P. infestans strains co-expressing a mitochondrially-localized mCherry fluorescent reporter"--Tomado del resumen