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Expresión multicasica de grupos de genes biosintéticos de cianobacterias y otras bacterias
Dipesh Dhakal1, Campbell W Eckhardt1, Yujia Jiang1
1Department of Medicinal Chemistry, Center for Natural Products, Drug Discovery and Development (CNPD3), University of Florida, Gainesville, Florida 32610, United States.
ACS synthetic biology
|September 2, 2025
Resumen
Los investigadores desarrollaron nuevos sistemas de expresión para los huéspedes microbianos, lo que permitió el descubrimiento de productos naturales (NP) a partir de grupos de genes biosintéticos bacterianos y cianobacterianos (BGC). Este avance facilita el descubrimiento de NP a través de diversos linajes bacterianos y cianobacterianos.
Área de la Ciencia:
- Microbiología
- Biología sintética
- Descubrimiento de productos naturales
Sus antecedentes:
- La expresión heteróloga de los grupos de genes biosintéticos (BGC) es crucial para el descubrimiento de productos naturales (NP).
- La expresión consistente a través de diversos huéspedes microbianos sigue siendo un desafío significativo.
- Los métodos existentes a menudo carecen de la versatilidad para acomodar BGC de varios linajes bacterianos, incluidas las cianobacterias.
Objetivo del estudio:
- Desarrollar sistemas vectoriales versátiles entre filas para expresar BGC bacterianas y cianobacterianas en múltiples huéspedes microbianos.
- Validar la eficacia de estos sistemas utilizando genes reporteros y BGC conocidos.
- Establecer plataformas sólidas para acelerar el descubrimiento de NP y comprender la función de BGC.
Principales métodos:
- Desarrollo de sistemas vectoriales entre filas compatibles con los huéspedes Gram-negativos (Escherichia coli), Gram-positivos (Bacillus subtilis) y cianobacterianos (Synechocystis PCC 6803, Anabaena sp. Se incluyen las siguientes categorías:
- Validación mediante expresión constitutiva e inducible de la proteína fluorescente amarilla mejorada (eYFP).
- Aplicación de vectores para expresar la shinorina y la violacea en las BGC, empleando sintonización de promotores, alimentación de sustratos, refactorización de BGC y control inducible.
Principales resultados:
- Expresión exitosa de BGC de cianobacterias y otras bacterias en E. coli, B. subtilis, Synechocystis PCC 6803 y Anabaena sp. Es el PCC 7120.
- Mejora de la producción de NP y mitigación de la toxicidad del huésped mediante estrategias de expresión optimizadas.
- Se ha demostrado la capacidad de Bacillus subtilis como huésped heterólogo para los BGC cianobacterianos.
Conclusiones:
- Los sistemas vectoriales de fila cruzada desarrollados proporcionan plataformas versátiles para la expresión heteróloga de BGC.
- Estos sistemas mejoran significativamente el potencial para el descubrimiento de nuevos productos naturales a partir de diversas fuentes bacterianas y cianobacterianas.
- Los hallazgos allanan el camino para aplicaciones más amplias en biología sintética y biotecnología microbiana.
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