At Chirotech, part of Dr. Reddy’s Laboratories Ltd., we have developed an efficient fermentation and downstream process for our recombinant E. coli-derived enzymes that is scalable from 1 litre up to multi-kilolitres. We use a combination of high throughput fermentation parameter screening with a Robolector® micro bioreactor system, coupled to parallel stirred tank fermentation capability (6 x 1 L) and up to 50 L in-house pilot scale production.
The resulting process maximises both the fermenter capacity, and the microorganism capability to produce extremely high levels of target enzyme. The fermentation process is highly scalable, robust and easily implemented at manufacturing scale. At 50 L, manufacturing of enzyme from glycerol stock of the strain through to a final dried enzyme powder can be completed within 7 days.
The process is a glucose fed-batch method able to support a high cell density culture that can achieve significant biomass levels of up to 80 g/L DCW and optical densities of up to 250. The downstream process is both simple and efficient, with a high recovery of enzyme, using few steps, to provide enzyme of sufficient quality to be directly used as a biocatalyst.
Examples will be given demonstrating the proficiency of this method of manufacture to achieve high enzyme productivity in an industrially relevant E. coli expression host.