Recombinant protein expression in E. coli is a fundamental process in various fields, withE. colibeing a key microbe for this purpose. This study focuses on a novel, scalable autoinduction method reliant on phosphate depletion to trigger protein expression inE. coli strains engineered for improved production. The method has shown significant enhancement in protein expression levels, reaching up to 55% of total cellular protein. Initial experiments in fed-batch fermentations achieved high cell densities (~30 gCDW/L) and notable protein titers (up to 8.1 ± 0.7 g/L). The methodology has been adapted for routine batch production in various culture volumes, from 20 μl to 100 ml, showcasing its versatility from high throughput screens to larger fermentations.
Key Points:
– Novel autoinduction method inE. coli using phosphate depletion for improved recombinant protein expression
– High expression levels achieved, reaching up to 55% of total cellular protein
– Scalable process demonstrated in fed-batch fermentations with high cell densities and protein titers
– Versatile method applicable from small-scale cultures to larger fermentations, offering broad utility in protein expression workflows
Tags: high throughput screening, process development, yeast, scale up, chromatography, formulation, bioreactor, cell culture
Read more on pmc.ncbi.nlm.nih.gov