Accelerating scale up of the ABUNDA® bioprocess

Challenge

As ENOUGH (formerly 3fBio) move mycoprotein production from their scale up facility in Glasgow to full scale production in the Netherlands, they wanted to design a suite of early diagnosis tests to determine when a batch of product is growing sub-optimally in the hope that issues can be mitigated and avoid reductions in the amount or quality of product.

At very large scale, a slight decrease in growth conditions can cause decreases in yields, and so this project was looking for warning signs of any issues with the growth of mycelium in ENOUGH’s product. Different conditions (such as nitrogen and oxygen limitation) were compared to identify if markers could be identified and a series of quick tests for nutrient limitation could be designed.

ABUNDA® mycoprotein

Solution

Transcriptomics is the analysis of all the RNA of a culture and provides a readout of the genes being expressed at any one time. Comparisons between lab scale and production scale allow issues in production scale to be identified and mitigated.

The team at ENOUGH performed lab scale fermentations under nutrient limitation and also took samples during large scale fermentation (15,000 L). RNA was extracted from these samples and sent to the University of Strathclyde for analysis.

Nick Tucker’s team then performed quality control analysis on the RNA and processed it RNA-Seq. The team sequenced the lab scale transcripts and one time course experiment from the large-scale fermentation. They also performed bioinformatic analysis to identify differentially expressed transcripts between the experiments allowing identification of biomarkers in the future.

Outcome 

Despite the challenges of Covid-19, which resulted in some samples being put in cold storage and not analysed during the project, a very detailed dataset was produced that has the potential to provide markers for input limitations. Nutrients were shown not to be in limitation during the large scale time course and mycotoxin pathways were not upregulated – allaying any potential safety concerns.