Farnberger, Judith E.; Lorenz, Elisabeth; Richter, Nina; Wendisch, Volker F.; Kroutil, Wolfgang: In vivo Plug-and-play: A Modular Multi-enzyme Single-cell Catalyst for the Asymmetric Amination of Ketoacids and Ketones. In: Microbial Cell Factories. Jg.16 H. 1. 2017
Inhalt
- Design and construction of modular plug-and-play single-cell biocatalysts
- Influence and toxicity of used substrates on the viability of E. coli W3110
- Fermentative amination of α-ketoacids
- Fermentative amination of α-ketoacids with metabolic NADH regeneration
- Fermentative reductive amination of α-ketoacids with non-metabolic NADH regeneration
- “In vitro-type” single-cell catalyzed amination of prochiral ketones
- Discussion
- Conclusions
- Methods
- Construction of bacterial strains and plasmids
- Construction of expression plasmids
- Construction of bacterial strains harbouring amino acid dehydrogenase genes
- Media and cultivation conditions
- Preparation of cell-free extracts
- Fermentative production of l-isoleucine and l-leucine
- Toxicity tests
- Activity assays
- Measurement of transaminase activity
- Measurement of activities of amino acid dehydrogenases
- Measurement of activities of NADH-recycling enzymes
- Biotransformations
- Analytical methods
- Authors’ contributions
- References