Precision fermentation with mass spectrometry‐based spent media analysis

Abstract

Optimization and monitoring of bioprocesses requires the measurement of several process parameters and quality attributes. Mass spectrometry (MS)‐based techniques such as those coupled to gas chromatography (GCMS) and liquid Chromatography (LCMS) enable the simultaneous measurement of hundreds of metabolites with high sensitivity. When applied to spent media, such metabolome analysis can help determine the sequence of substrate uptake and metabolite secretion, consequently facilitating better design of initial media and feeding strategy. Furthermore, the analysis of metabolite diversity and abundance from spent media will aid the determination of metabolic phases of the culture and the identification of metabolites as surrogate markers for product titer and quality. This review covers the recent advances in metabolomics analysis applied to the development and monitoring of bioprocesses. In this regard, we recommend a stepwise workflow and guidelines that a bioprocesses engineer can adopt to develop and optimize a fermentation process using spent media analysis. Finally, we show examples of how the use of MS can revolutionize the design and monitoring of bioprocesses.

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

“…Cell culture media is a key component of industrial bioprocess that is immediately disposed of as waste after the primary fermentation. It is well described that many nutrients remain within media post-primary culture, 27–29 and this is shown in the metabolomic, proteomic and elemental analysis of the two media types used here, that we term CHO spent media (CSM) and fungal spent media (FSM) (Fig. 4–6).…”

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

“…Cell culture media is a key component of industrial bioprocess that is immediately disposed of as waste after the primary fermentation. It is well described that many nutrients remain within media post-primary culture (Wang et al ., 2017; O’Neill et al ., 2022; Dodia et al ., 2023), and this is shown in the metabolomic, proteomic and elemental analysis of the two media types used here, that we term CHO spent media (CSM) and fungal spent media (FSM) (figures 4-6).…”

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

“…From this point, an 18.5 h expression was carried out with the temperature dropped to 30 • C and left overnight. At the end of the expression, three 5 mL technical replicate samples were taken for protein purification and yield measurement by centrifuging the samples at 4000× g for 20 min and discarding the supernatant, and then the resulting biomass pellet was frozen at −80 • C overnight and stored at −20 • C until purification could be carried out [26,27].…”