Browsing by Author "Poblete Castro, Ignacio"

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    Co-synthesis of medium-chain-length polyhydroxyalkanoates and CdS quantum dots nanoparticles in Pseudomonas putida KT2440
    (2017) Oliva Arancibia, Bárbara; Ordenes Aenishanslins, Nicolás; Bruna, Nicolás; Ibarra, Paula S.; Zacconi, Flavia C. M.; Pérez Donoso, José M.; Poblete Castro, Ignacio
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    Discovery of New Phenylacetone Monooxygenase Variants for the Development of Substituted Indigoids through Biocatalysis
    (2022) Núñez Navarro, Nicolás Ernesto; Salazar Muñoz, Javier Alonso; Castillo Suzarte, Francisco Javier; Ramírez Sarmiento, César Antonio; Poblete Castro, Ignacio; Zacconi, Flavia C. M.; Parra, Loreto
    Indigoids are natural pigments obtained from plants by ancient cultures. Romans used them mainly as dyes, whereas Asian cultures applied these compounds as treatment agents for several diseases. In the modern era, the chemical industry has made it possible to identify and develop synthetic routes to obtain them from petroleum derivatives. However, these processes require high temperatures and pressures and large amounts of solvents, acids, and alkali agents. Thus, enzyme engineering and the development of bacteria as whole-cell biocatalysts emerges as a promising green alternative to avoid the use of these hazardous materials and consequently prevent toxic waste generation. In this research, we obtained two novel variants of phenylacetone monooxygenase (PAMO) by iterative saturation mutagenesis. Heterologous expression of these two enzymes, called PAMOHPCD and PAMOHPED, in E. coli was serendipitously found to produce indigoids. These interesting results encourage us to characterize the thermal stability and enzyme kinetics of these new variants and to evaluate indigo and indirubin production in a whole-cell system by HPLC. The highest yields were obtained with PAMOHPCD supplemented with L-tryptophan, producing ~3000 mg/L indigo and ~130.0 mg/L indirubin. Additionally, both enzymes could oxidize and produce several indigo derivatives from substituted indoles, with PAMOHPCD being able to produce the well-known Tyrian purple. Our results indicate that the PAMO variants described herein have potential application in the textile, pharmaceutics, and semiconductors industries, prompting the use of environmentally friendly strategies to obtain a diverse variety of indigoids.
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    Exploiting the natural poly(3-hydroxyalkanoates) production capacity of Antarctic Pseudomonas strains : from unique phenotypes to novel biopolymers
    (2019) Pacheco, Nicolás; Orellana Sáez, Matias; Pepczynska, Marzena; Enrione, Javier I.; Bassas Galia, Mónica; Borrero de Acuña, José Manuel; Zacconi, Flavia C. M.; Marcoleta, Andrés Esteban; Poblete Castro, Ignacio