Synthetic Multienzyme Complexes Assembled on Virus-like Particles for Cascade Biosynthesis In Cellulo.
Bioconjug Chem
; 31(10): 2413-2420, 2020 10 21.
Article
en En
| MEDLINE
| ID: mdl-33001630
ABSTRACT
Multienzyme complexes, or metabolons, are natural assemblies or clusters of sequential enzymes in biosynthesis. Spatial proximity of the enzyme active sites results in a substrate channeling effect, streamlines the cascade reaction, and increases the overall efficiency of the metabolic pathway. Engineers have constructed synthetic multienzyme complexes to acquire better control of the metabolic flux and a higher titer of the target product. As most of these complexes are assembled through orthogonal interactions or bioconjugation reactions, the number of enzymes to be assembled is limited by the number of orthogonal interaction or reaction pairs. Here, we utilized the Tobacco mosaic virus (TMV) virus-like particle (VLP) as protein scaffold and orthogonal reactive protein pairs (SpyCatcher/SpyTag and SnoopCatcher/SnoopTag) as linker modules to assemble three terpene biosynthetic enzymes in Escherichia coli. The enzyme assembly switched on the production of amorpha-4,11-diene, whereas the product was undetectable in all the controls without assembly. This work demonstrates a facile strategy for constructing scaffolded catalytic nanomachineries to biosynthesize valuable metabolites in bacterial cells, and a unique assembly induced the switch-on mechanism in biosynthesis for the first time.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Terpenos
/
Virus del Mosaico del Tabaco
/
Virión
/
Escherichia coli
/
Complejos Multienzimáticos
Idioma:
En
Revista:
Bioconjug Chem
Asunto de la revista:
BIOQUIMICA
Año:
2020
Tipo del documento:
Article
País de afiliación:
China