Small-scale mashing procedure for predicting ethanol yield of sorghum grain

A small-scale mashing (SSM) procedure requiring only 300 mg of samples was investigated as a possible method of predicting ethanol yield of sorghum grain. The initial SSM procedure, which was conducted similarly to the mashing step in a traditional fermentation test, hydrolyzed just 38.5–47.2% of total sorghum starch to glucose. The initial procedure was simplified to contain only one liquefaction step, which did not influence subsequent saccharification. Thereafter, parameters such as temperature, pH, enzyme dosage, and saccharification time were optimized. Results showed that 91.2–97.5% of the total starch in 18 sorghum hybrids had been hydrolyzed to glucose using the following conditions: liquefaction at 86 °C for 90 min, 20 μL of α-amylase per 30 g of sample; pH adjustment by adding 50 μL of 2 M acetate buffer at pH 4.2 to each microtube; saccharification at 68 °C for 90 min, 200 μL of amyloglucosidase per 30 g of sample. There were strong linear correlations between completely hydrolyzed starch (CHS) from SSM and ethanol yields from both traditional (R2 = 0.86) and simultaneous saccharification and fermentation (SSF, R2 = 0.93) procedures. CHS was a better indicator for predicting ethanol yield in fermentation than total starch.