Fig. 6.

Hydrolysis of avicel or PASC and production of glucose or ethanol by the engineered cellulosome. (A) Analysis of glucose release from avicel or PASC. The cells were precultured and 10 OD cells were inoculated into YP medium supplemented with 1% avicel or PASC with or without 100 mM methylglyoxal (MG). In the presence of MG, the glucose metabolism of host cells was arrested, while the glucose produced during the hydrolysis remained intact. (B) Optimization of the ratio of EH and the SH (with OlpB-ScGPI and CipA1B9C). The cells were precultured in YPG medium and washed with PBS twice then inoculated at different ratio in YP media and supplemented with 0.042% Tween 80 and 1% avicel. Fermentation was carried out at 37 °C at 300 rpm for 4 d and ethanol concentration was analyzed using HPLC. (C and D) Time course of cellulosic ethanol production by yeast consortium composed of EH (EH-P1-44) and either SH-1B3C, SH-1B6C, SH-0B9C, SH-1B9C, or SH-2B9C at the optimized ratio of 10:1. Ethanol production from 10 g/L avicel (C) and 20 g/L PASC (D) was analyzed using HPLC.