Cascade Synthesis of l-Homoserine Catalyzed by Lyophilized Whole Cells Containing Transaminase and Aldolase Activities: The Mathematical Modeling Approach

Aldolase and transaminase coexpressed in Escherichia coli cells and lyophilized (i.e., lyophilized whole-cell biocatalyst (LWCB)) were used as biocatalysts for the one-pot cascade synthesis of l-homoserine with substrate cycling. The kinetic analysis of enzymes within lyophilized cells was performed...

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Detalles Bibliográficos
Autores: Česnik, Morana, Sudar, Martina, Hernández, Karel, Qi, Yuyin, Charnock, Simon J., Vasić-Rački, Đurdica, Clapés Saborit, Pere, Blažević, Zvjezdana Findrik
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/253955
Acceso en línea:http://hdl.handle.net/10261/253955
Access Level:acceso abierto
Palabra clave:Aldolases
Mathematical Modeling Approach
Descripción
Sumario:Aldolase and transaminase coexpressed in Escherichia coli cells and lyophilized (i.e., lyophilized whole-cell biocatalyst (LWCB)) were used as biocatalysts for the one-pot cascade synthesis of l-homoserine with substrate cycling. The kinetic analysis of enzymes within lyophilized cells was performed to evaluate the behavior of the system. The best result among the performed fed-batch reactor experiments achieved was 640.8 mM (76.3 g L–1) of l-homoserine with a volume productivity of 2.6 g L–1 h–1. This is comparable with the results of the same cascade synthesis using cell-free extracts (CFEs) and significantly better than the reports in the literature applying fermentation technology. The approach applied here can serve as guidance for the design of microbial cells with an optimal ratio of expressed enzymes that act as biocatalysts in the cascade, resulting in lower biocatalyst cost, no need for the addition of expensive coenzymes, and enhanced enzyme stability as compared with cell-free extracts.