Redesigning Spent Media from Cell Culture Bioprocess to Feed New Bacterial Fermentations

Lynch, Ciara; Jordan, Lynda; Connell, David J O’

doi:10.1007/978-3-030-79871-0_5

Cited by 3 publications

(3 citation statements)

References 59 publications

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“…The high lactic acid levels in the FSM therefore aids in explaining the reduced protein yield obtained in the flask cultures in this media (0.216 g/L as opposed to 0.489 g/L in LB). There is precedence as it has been shown before that host cell protein (HCP) or leftover metabolite presence can affect the growth of cells in secondary cultures (Hsiao, Glatz and Glatz, 1994; Wu, Ruan and Lam, 1998; Lowrey, Armenta and Brooks, 2016; Lynch, Jordan and J O’ Connell, 2021). In the shake flask cultures of E. coli fed with either CHO spent media (CSM) or fungal spent media (FSM), there was a noticeable decrease in both growth rate and protein yield compared to the cells grown on nutrient rich LB media (Table 1).…”

Section: Discussionmentioning

confidence: 99%

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

Lynch,

Cerrone,

O’Connor

et al. 2023

Preprint

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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.

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Section: Discussionmentioning

confidence: 99%

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

Lynch,

Cerrone,

O’Connor

et al. 2023

Preprint

Self Cite

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show abstract

“…There is precedence as it has been shown before that host cell protein (HCP) or leover metabolite presence can affect the growth of cells in secondary cultures. 15,17,39,40 In the shake ask cultures of E. coli fed with either CHO spent media (CSM) or fungal spent media (FSM), there was a noticeable decrease in both growth rate and protein yield compared to the cells grown on nutrient rich LB media (Table 1). This is likely the result of the reduced nutrient value of the media post-primary culture, though high lactic acid concentration had an effect on the FSM-fed secondary cultures at shake ask scale.…”

Section: Cho Spent Media (Csm) Supports Secondary Culturesmentioning

confidence: 99%

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

Lynch,

Cerrone,

O'Connor

et al. 2024

RSC Sustain.

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“…While a prominent goal in sustainability practices is to reduce the volume of waste that is produced by industries and processes, dealing with the large amounts of waste currently produced by industrial processes is where the circular bioeconomy can have an immediate impact. One process-intensive area producing significant volumes of waste not currently reused is in the production of biopharmaceuticals, which generates significant volumes of cell culture media waste [3]. A study into the life cycle of these media found that their cost dominated the operating cost of monoclonal antibody production, particularly with regard to continuous culturing methods such as perfusion due to the higher consumption of media in this bioprocess system [4].…”

Section: Introductionmentioning

confidence: 99%

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

Murphy,

Lynch,

O’Connell

2024

Applied Microbiology

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Spent fermentation media from bioprocessing represent a significant waste stream, and interest in recycling them as part of the developing circular bioeconomy is growing. The potential to reuse yeast spent culture media (YSM) to feed secondary bacterial fermentations producing recombinant protein was investigated in this study. Elemental and amino acid compositional analysis using inductively coupled plasma mass spectrometry (ICP-MS) and LC-MS/MS identified significant differences in the concentrations of 6 elements and 18/20 amino acids in YSM compared with rich microbiological media (LB). Restoration of levels of magnesium and sodium through addition of their salts and amino acids from tryptone supplementation led to the expression of equivalent titres of recombinant proteins by E. coli (0.275 g/L), compared to that in LB media (0.296 g/L) and BMMY media (0.294 g/L) in shake flask culture. When this supplementation strategy was employed in a bioreactor system, we observed a significant increase in recombinant protein titre using the supplemented YSM (2.29 (±0.02) g/L) over that produced using LB media (1.29 (±0.09) g/L). This study demonstrates through highly sensitive compositional analysis and identification of supplementation strategies the potential to valorise spent media from yeast fermentations that underpin industrial processes of significant scale, creating a circular approach to waste stream management.

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Redesigning Spent Media from Cell Culture Bioprocess to Feed New Bacterial Fermentations

Cited by 3 publications

References 59 publications

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

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

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

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

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