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Biosíntesis dirigida por precursores de macrolides de 16 miembros por la eritromicina policetida sintasa
K Kinoshita1, P G Williard, C Khosla
1Department of Chemistry, Box H, Brown University, Providence, Rhode Island 02912-9108, USA.
Journal of the American Chemical Society
|July 18, 2001
Resumen
La 6-deoxyeritronolide B sintasa mutante en Streptomyces coelicolor aceptó un análogo de triketida, formando un octaketida macrolactona. Esto revela conocimientos sobre la especificidad del sustrato de la policetida sintasa y la nueva biosíntesis de macrolactona.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Biología Molecular Biología Molecular
- Biología sintética Biología sintética.
Sus antecedentes:
- Streptomyces coelicolor CH999/pJRJ2 contiene una 6-deoxyeritronolide B sintasa mutante (DEBS) con un sitio activo bloqueado de la cetosintasa (KS1).
- Esta mutación impide la biosíntesis natural del 6-deoxyeritronolide B, un policetido clave.
Objetivo del estudio:
- Para investigar la especificidad del sustrato de la enzima mutante DEBS.
- Explorar el potencial de una nueva biosíntesis de macrolactona utilizando las sintetasas poliquetidas de ingeniería.
Principales métodos:
- Administración de análogos sintéticos de triketida a cultivos de S. coelicolor que expresan el mutante DEBS.
- Aislamiento y caracterización de los productos de macrolactona resultantes utilizando técnicas analíticas.
Principales resultados:
- La incubación con un análogo específico insaturado de triketida (6) produjo un nuevo octaketida de 16 miembros macrolactona (33).
- Esto indica que el DEBS mutante procesó el triketido como un análogo del diketido, involucrando el módulo 2.
- Otros análogos (18, 31, 32) también produjeron macrolactonas correspondientes (34, 35, 36), confirmando la aceptación del sustrato.
- Un análogo C-2 desmetilado (10) no pudo producir una macrolactona, lo que pone de relieve la importancia de las características estructurales específicas.
Conclusiones:
- La enzima DEBS mutada exhibe una especificidad alterada del sustrato, aceptando análogos de triketida.
- Este sistema de ingeniería permite la síntesis de novo de nuevas macrolactonas.
- El estudio proporciona información valiosa sobre los mecanismos de alargamiento de la cadena poliquetídica y la ingeniería enzimática para la biosíntesis.
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