In the SSF process, fermentation media was a 50 mM sodium acetate solution (at various pH) containing 5 g/L yeast extract and 10 g/L peptone. One gram of lipid-extracted biomass was added into a reaction bottle and reaction volume was maintained at 40 mL. S. cerevisiae in active dry granule was added into 39 C autoclaved distilled water to make yeast solution at various concentrations (12.5e100 g/L). 1 mL of amyloglucosidase at different concentration (15e75 units/mL) and 2 mL of yeast solution were added simultaneously to start the reaction. At certain interval time, 0.5 mL of samples were taken and centrifuged at 10,000 g for 5 min. Supernatant of the samples were analysed for sugar and bioethanol content. In this regard, the reaction time of 360 min was taken as total observation time.