<i>In situ</i> solid-liquid extraction enhances recovery of taxadiene from engineered <i>S. cerevisiae</i> cell factories

Santoyo‐Garcia, Jorge H.; Walls, Laura E.; Nowrouzi, Behnaz; Ochoa-Villareal, Marisol; Loake, Gary J.; Dimartino, Simone; Rios‐Solis, Leonardo

doi:10.1101/2021.07.12.452013

Cited by 1 publication

(1 citation statement)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This suggested that the integration of additional genes resulted in a decreased cellular fitness. In addition, the LRS5 strain was recently found to have a reduced mechanical stress tolerance compared to the wild-type, CEN.PK-1C, strain [40]. In the presence of an in situ solid phase adsorbent resin with a concentration of 12% w/v, the growth of LRS5 was completely inhibited, whilst the growth of the parental CEN.PK-1C was not significantly affected, further suggesting that metabolic engineering has a detrimental effect on cellular fitness and stress tolerance.…”

Section: Quorum Sensing In S Cerevisiaementioning

confidence: 99%

Enhancing Saccharomyces cerevisiae Taxane Biosynthesis and Overcoming Nutritional Stress-Induced Pseudohyphal Growth

2022

Self Cite

View full text Add to dashboard Cite

The recent technological advancements in synthetic biology have demonstrated the extensive potential socio-economic benefits at laboratory scale. However, translations of such technologies to industrial scale fermentations remains a major bottleneck. The existence and lack of understanding of the major discrepancies in cultivation conditions between scales often leads to the selection of suboptimal bioprocessing conditions, crippling industrial scale productivity. In this study, strategic design of experiments approaches were coupled with state-of-the-art bioreactor tools to characterize and overcome nutritional stress for the enhanced production of precursors to the blockbuster chemotherapy drug, Taxol, in S. cerevisiae cell factories. The batch-to-batch variation in yeast extract composition was found to trigger nutritional stress at a mini-bioreactor scale, resulting in profound changes in cellular morphology and the inhibition of taxane production. The cells shifted from the typical budding morphology into striking pseudohyphal cells. Doubling initial yeast extract and peptone concentrations (2×YP) delayed filamentous growth, and taxane accumulation improved to 108 mg/L. Through coupling a statistical definitive screening design approach with the state-of-the-art high-throughput micro-bioreactors, the total taxane titers were improved a further two-fold, compared to the 2×YP culture, to 229 mg/L. Filamentous growth was absent in nutrient-limited microscale cultures, underlining the complex and multifactorial nature of yeast stress responses. Validation of the optimal microscale conditions in 1L bioreactors successfully alleviated nutritional stress and improved the titers to 387 mg/L. Production of the key Taxol precursor, T5αAc, was improved two-fold to 22 mg/L compared to previous maxima. The present study highlights the importance of following an interdisciplinary approach combining synthetic biology and bioprocessing technologies for effective process optimization and scale-up.

show abstract

Section: Quorum Sensing In S Cerevisiaementioning

confidence: 99%