Grass, a low-cost lignocellulosic feedstock with relatively low lignin content, serves as a potential carbon source for microbial processes due to its polymeric sugar content. However, converting carbohydrates into monomeric sugars presents challenges due to complex lignocellulosic matrix. In this study, a mixture of perennial ryegrass and white clover (RG) was pretreated with alkaline before being enzymatically hydrolysed for use as a fermentable sugar source for the oleaginous yeast Metschnikowia pulcherrima. The dilute alkaline pretreatment approach resulted in a 3 times improvement in the conversion of RG, yielding a fermentable sugar concentration of 56.5 g/L. The yeast exhibited a growth yield (Ym/m) of 0.47, producing 23.7 g/L of dry cell weight at 20°C over 140 hours, with a lipid content of 37 % with a similar composition to high oleic palm oil. These findings suggest that RG can be utilised for formulating an oleaginous yeast medium containing the necessary nutrients.
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