Chamizo Ampudia, Alejandro

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https://academica-e.unavarra.es/handle/2454/48847

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Chamizo Ampudia

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Alejandro

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Ciencias del Medio Natural

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0000-0002-0563-7954

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752002

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811497

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2024 - 20252
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    PublicationOpen Access
    The effectiveness of polyhydroxyalkanoate (PHA) extraction methods in gram-negative pseudomonas putida U
    (MDPI, 2025-01-09) Getino, Luis; García, Irene; Cornejo Ibergallartu, Alfonso; Mateos, Raúl; Ariza-Carmona, Luisa M.; Sánchez-Castro, Natalia; Morán Juez, José Fernando; Olivera, Elías R.; Chamizo Ampudia, Alejandro; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Institute for Advanced Materials and Mathematics - INAMAT2
    Bioplastics are emerging as a promising solution to reduce pollution caused by petroleum-based plastics. Among them, polyhydroxyalkanoates (PHAs) stand out as viable biotechnological alternatives, though their commercialization is limited by expensive downstream processes. Traditional PHA extraction methods often involve toxic solvents and high energy consumption, underscoring the need for more sustainable approaches. This study evaluated physical and chemical methods to extract PHAs from Pseudomonas putida U, a bacterium known to produce poly-3-hydroxyoctanoate P(3HO). Lyophilized cells underwent six extraction methods, including the use of the following: boiling, sonication, sodium hypochlorite (NaClO), sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and chloroform. Physical methods such as boiling and sonication achieved yields of 70% and 60%, respectively, but P(3HO) recovery remained low (30–40%). NaClO extraction provided higher yields (80%) but resulted in significant impurities (70%). NaOH methods offered moderate yields (50–80%), with P(3HO) purities between 50% and 70%, depending on the conditions. Spectroscopic and analytical techniques (FTIR, TGA, NMR, GPC) identified 0.05 M NaOH at 60 °C as the optimal extraction condition, delivering high P(3HO) purity while minimizing environmental impact. This positions NaOH as a sustainable alternative to traditional halogenated solvents, paving the way for more eco-friendly PHA production processes.
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    PublicationOpen Access
    A new oxidative pathway of nitric oxide production from oximes in plants
    (Cell Press, 2024) López Gómez, Pedro; Buezo Bravo, Javier; Urra Rodríguez, Marina; Cornejo Ibergallartu, Alfonso; Esteban Terradillos, Raquel; Fernández de los Reyes, Jorge; Urarte Rodríguez, Estíbaliz; Rodríguez-Dobreva, Estefanía; Chamizo Ampudia, Alejandro; Eguaras, Alejandro; Wolf, Sebastian; Marino Bilbao, Daniel; Martínez Merino, Víctor; Morán Juez, José Fernando; Ciencias; Zientziak; Institute for Multidisciplinary Research in Applied Biology - IMAB; Institute for Advanced Materials and Mathematics - INAMAT2
    Nitric oxide (NO) is an essential reactive oxygen species and a signal molecule in plants. Although several studies have proposed the occurrence of oxidative NO production, only reductive routes for NO production, such as the nitrate (NO-3) -upper-reductase pathway, have been evidenced to date in land plants. However, plants grown axenically with ammonium as the sole source of nitrogen exhibit contents of nitrite and NO3, evidencing the existence of a metabolic pathway for oxidative production of NO. We hypothesized that ox- imes, such as indole-3-acetaldoxime (IAOx), a precursor to indole-3-acetic acid, are intermediate oxidation products in NO synthesis. We detected the production of NO from IAOx and other oximes catalyzed by peroxidase (POD) enzyme using both 4-amino-5-methylamino-20,70-difluorescein fluorescence and chem- iluminescence. Flavins stimulated the reaction, while superoxide dismutase inhibited it. Interestingly, mouse NO synthase can also use IAOx to produce NO at a lower rate than POD. We provided a full mech- anism for POD-dependent NO production from IAOx consistent with the experimental data and supported by density functional theory calculations. We showed that the addition of IAOx to extracts from Medicago truncatula increased the in vitro production of NO, while in vivo supplementation of IAOx and other oximes increased the number of lateral roots, as shown for NO donors, and a more than 10-fold increase in IAOx dehydratase expression. Furthermore, we found that in vivo supplementation of IAOx increased NO pro- duction in Arabidopsis thaliana wild-type plants, while prx33-34 mutant plants, defective in POD33-34, had reduced production. Our data show that the release of NO by IAOx, as well as its auxinic effect, explain the superroot phenotype. Collectively, our study reveals that plants produce NO utilizing diverse molecules such as oximes, POD, and flavins, which are widely distributed in the plant kingdom, thus intro- ducing a long-awaited oxidative pathway to NO production in plants. This knowledge has essential impli- cations for understanding signaling in biological systems.