Transport-controlled growth decoupling for self-induced protein expression with a glycerol-repressible genetic circuit
Küçük Resim Yok
Tarih
2024
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Wiley
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Decoupling cell formation from recombinant protein synthesis is a potent strategy to intensify bioprocesses. Escherichia coli strains with mutations in the glucose uptake components lack catabolite repression, display low growth rate, no overflow metabolism, and high recombinant protein yields. Fast growth rates were promoted by the simultaneous consumption of glucose and glycerol, and this was followed by a phase of slow growth, when only glucose remained in the medium. A glycerol-repressible genetic circuit was designed to autonomously induce recombinant protein expression. The engineered strain bearing the genetic circuit was cultured in 3.9 g L-1 glycerol + 18 g L-1 glucose in microbioreactors with online oxygen transfer rate monitoring. The growth was fast during the simultaneous consumption of both carbon sources (C-sources), while expression of the recombinant protein was low. When glycerol was depleted, the growth rate decreased, and the specific fluorescence reached values 17% higher than those obtained with a strong constitutive promoter. Despite the relatively high amount of C-source used, no oxygen limitation was observed. The proposed approach eliminates the need for the substrate feeding or inducers addition and is set as a simple batch culture while mimicking fed-batch performance. The authors propose a strategy that combines cell engineering and genetic circuit design to decouple growth rate from recombinant protein expression based on the carbon source transport. A simple batch scheme allows autoinduction and growth rate shift, mimicking a fed-batch mode. The results are potentially useful for product and process development. image
Açıklama
Anahtar Kelimeler
Bioprocess İntensification, C-Source Mixtures, Genetic Circuit, Growth Decoupling, Microbial Engineering, Toggle Switch, Glucose, Metabolism, System
Kaynak
Biotechnology and Bioengineering
WoS Q Değeri
N/A
Scopus Q Değeri
Q1
