Monitoring growth and lipid production of new isolated oleaginous yeast Cryptococcus aerius UIMC65 on glucose and xylose cultures

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

“…Specific fluorescent molecules emit higher fluorescence depending on cellular physiological parameters. For example, cellular pH can be detected using BCECF, and SNARF‐1; membrane potential can be measured using DHR 123, DiBAC, DiOC 6 (3), Oxonol, TMRM and Rhodamine 123 (specific for mitochondrial membrane); enzyme activities can be detected using Carboxyfluoresceindiacetate (CFDA) (related to esterase activity), ELF 97‐phosphate (related to alkaline phosphatase activity), Calcein‐AM, CFSE; Respiratory activity, glucose uptake and reactive oxygen species (ROS) can be measured using CTC, 2‐NBDG, and DHR/DHE, respectively . In most of the microbial bioengineering experiments, FITC has been used to tag proteins to study expression and activities.…”

“…But, fluorescent dyes related to lipids require addition of solvents or robust pre‐treatment conditions that can facilitate the penetration of fluorescent dyes into the cell, and bind to the lipid bodies. Earlier, phosphate buffer saline (PBS) was used for pre‐treatment of oleaginous microorganisms prior to staining with lipophilic dyes Sudan Black B and Nile Red . A study by Kimura et al .…”

“…This may be due to the availability of genetic information on Y. lipolytica . One of the advantages of oleaginous yeast strains is that they can accumulate intracellular lipids and utilize lignocellulose materials for growth and cell development as well, simultaneously . Recent reviews by Athenaki et al, Chaturvedi et al, Xue et al .…”

“…coupled a bioreactor to online flow cytometry (Accuri C6), and online continuous observation carried out for variation in cell shape, growth dynamics and intracellular lipid accumulation. Overall, during fermentation, variations in yeast cell physiological properties in relation to lipid biosynthesis can be studied using flow cytometry …”

“…27 One of the advantages of oleaginous yeast strains is that they can accumulate intracellular lipids and utilize lignocellulose materials for growth and cell development as well, simultaneously. 28 Recent reviews by Athenaki et al, 29 Chaturvedi et al, 30 Xue et al 31 discussed the physiology of oleaginous yeasts, and their lipids production capability, as well potential biotechnological applications and constraints. It has been reported that Rhodosporidium toruloides and Lipomyces starkeyi can use xylose as a carbon source for cellular growth and can accumulate lipids as well.…”