Feeding secondary fermentations with mammalian and fungal culture waste streams increases productivity and resource efficiency

Abstract

The evolution of the circular bioeconomy will require the realisation of new value from waste streams generated from all manufacturing processes. Bioprocessing of recombinant proteins and functional foods using animal and microbial fermentations are fast growing industries with increasing volumes of spent culture media waste resulting in significant resource inefficiency. Here Chinese hamster ovary cell spent media (CSM) and Trametes versicolor fungal cell spent media (FSM) were used as model waste streams to feed Escherichia coli in secondary fermentations. Expression of a recombinant fusion protein was used to measure the value of these streams as feedstuff. E. coli cultures fed with either waste stream in bioreactors produced equivalent yields of the protein as those fed with rich microbiological media. Separately CSM was tested as a replacement feed for the fungal cell fermentation producing beta and alpha glucans. This waste-fed fermentation resulted in a 2-fold higher glucan yield than when using standard corn steep liquor feed. Quantitative analysis of the spent media content with mass spectrometry and hplc methods showed significant differences in elemental composition profiles, quantities of individual amino acids and carboxylic acids and levels of carbon sources. These findings underline the versatility of E.coli in utilisation of waste media as a feedstuff and show that the range of applications of CHO cell culture media waste are not limited to feeding bacterial fermentation. Exploration of cellular bioprocesses that can efficiently use cell media waste as fermentation feed will be a valuable step towards increased resource efficiency in bioprocessing industries.

Valorisation of Spent Yeast Fermentation Media through Compositional-Analysis-Directed Supplementation

“…However, the impact of supplementation with magnesium and tryptone together on recombinant protein expression in the bioreactor was not expected with a high titre of 2.29 (±0.02) g/L, compared to a titre of 1.29 (±0.09) g/L from cultures fed with LB (Figure 3B). The percent recombinant protein in grams per gram of cell dry weight obtained for YSM plus magnesium-fed culture (18.83%) was equivalent to LB (18.12%), albeit with a much lower CDW of 4.2 vs. 7.1 g/L [8]. The same measurement showed improved performance of the magnesium-and tryptone-supplemented YSM, with an increase of almost 2%, 19.94% per gram of CDW, over that of LB (Figure 3C).…”

“…The WARIEN values for continuous production of recombinant proteins in industry could be substantially reduced by reusing waste media, as media have been shown to be the largest contributor to the high WARIEN values associated with this type of bioprocess, with around 95% of the PMI for water being media-related [7]. The feasibility of this circular model has already been demonstrated with both mammalian and fungal spent cell media, both of which achieved equivalent levels of productivity in a secondary bacterial culture as the standard rich medium, lysogeny broth (LB) [8,9].…”