Online flow cytometry, an interesting investigation process for monitoring lipid accumulation, dimorphism, and cells\u2019 growth in the oleaginous yeast Yarrowia lipolytica JMY 775

Abstract This study aims to understand and better control the main biological mechanisms and parameters modulating the various phenomena affecting Yarrowia lipolytica JMY775 and its lipids accumulation. The results obtained in this study stress forward that the use of an original tool, consisting of coupling bioreactors to online flow cytometry, is highly efficient. Throughout 48\ua0h of culturing, this emerging process allowed an online continuous observation of the effects of pH and/or aeration on the cell growth and dimorphism and lipid accumulation by Y. lipolytica . This present study showed clearly that online flow cytometry is an advantageous tool for the real-time monitoring of microbial culture at a single-cell level. Indeed, the present investigation showed for the first time that profiling of the various phenomena and their monitoring upon culture time is now possible by coupling online cytometry with culture bioreactors

pH level has a strong impact on population dynamics of the yeast Yarrowia lipolytica and oil micro-droplets in multiphasic bioreactor

The oleaginous yeast Yarrowia lipolytica has the ability to use oils and fats as carbon source, making it a promising cell factory for the design of alternative bioprocesses based on renewable substrates. However, such a multiphasic bioreactor design is rather complex and leads to several constraints when considering emulsification of the oil-in-water mixture, foaming and cell growth/physiology on hydrophobic substrate. This study aims to shed light on the effect of pH changes on the physico-chemical properties of the cultivation medium and on cell physiology. It was indeed observed that at a pH value of 6, cell growth rate and intracellular lipid accumulation were optimized. Additionally, foaming was significantly reduced. In order to avoid over foaming in bioreactor, without impairing cell physiology, the use of alternative processes that can only act on the physical structure of culture medium, seems to be an effective alternative to usual chemical anti-foam agents

Strategies at Bioreactor Scale for the Production of Recombinant Proteins in Yarrowia lipolytica

Recombinant protein production represents a multibillion-dollar market. Therefore, it constitutes an important research field both in academia and industry. The use of yeast as cell factory presents several advantages such as ease of genetic manipulation, growth at high cell density, and possibility of posttranslational modifications. Yarrowia lipolytica is considered as one of the most attractive hosts due to its ability to metabolize raw substrate, to express genes at high level, and to secrete protein in large amounts. In the recent years, several reviews were dedicated to genetic tools developed for this purpose. Although the construction of efficient cell factory for recombinant protein synthesis is important, the development of an efficient process for protein production constitutes an equally vital aspect. Indeed, a sports car could not drive fast on a gravel road. The aim of this review is to provide a comprehensive snapshot of process tools to consider for recombinant protein production in bioreactor using Y. lipolytica as a cell factory, in order to facilitate the decision-making for future strain and process engineering