Protocols for Monitoring Growth and Lipid Accumulation in Oleaginous Yeasts
Oleaginous yeasts can synthesize and store lipids up to 20% of their dry weight and have emerged as resources of choice for biotechnological applications, such as bio-lipid production. The number of species and mutant libraries consequently available for screening is exponentially growing. Cultivation strategies and growth media for bio-lipid production need to be optimized to accelerate screening and identification of production strains. In this chapter we describe methods for high-throughput cell growth in 96 microtiter plates in various media including opaque broth by using a fluorescent reporter, carbon/nitrogen ratio determination for optimal lipid accumulation, and in vivo real-time detection of lipid accumulation using a neutral lipid fluorescent dye. We provide examples using two well-established oleaginous yeasts, Yarrowia lipolytica and Rhodosporidium toruloides. These methods can be extended to other oleaginous yeast species for high-throughput screening of bio-lipid accumulation.
Keywords:Lipid accumulation Oleaginous yeast Rhodosporidium Single cell oil Yarrowia
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