Date of Award
Doctor of Philosophy
Civil, Construction, and Environmental Engineering
Hans Van Leeuwen
Anthony L. Pometto Iii
The use of bio-based feedstock to support an economy based on renewable resources is becoming extremely important for generating biofuels and biobased products to reduce nation's dependency on imported petroleum fuels. This research aims at developing a biorefinery platform to convert corn-ethanol co-product, corn fiber, into fermentable sugars at a lower temperature with minimal use of chemicals. White-rot (Phanerochaete chrysosporium), brown-rot (Gloeophyllum trabeum) and soft-rot (Trichoderma reesei) fungi were used in this research to biologically break down cellulosic and hemicellulosic components of corn fiber into fermentable sugars. Laboratory-scale simultaneous saccharification and fermentation (SSF) process proceeded by in-situ cellulolytic enzyme induction enhanced overall enzymatic hydrolysis of hemi/cellulose from corn fiber into simple sugars (mono-, di-, tri-saccharides). The yeast fermentation of hydrolyzate yielded 7.1, 8.6 and 4.1 g ethanol per 100 g corn fiber when saccharified with the white-, brown-, and soft-rot fungi, respectively. The highest corn-to-ethanol yield (8.6 g ethanol/ 100 g corn fiber) was equivalent to 42 % of the theoretical ethanol yield from starch and cellulose in corn fiber. Cellulase, xylanase and amylase activities of these fungi were also investigated over a week long solid-substrate fermentation of corn fiber. G. trabeum had the highest activities for starch (160 mg glucose/mg protein.min) and on day three of solid-substrate fermentation. P. chrysosporium had the highest activity for xylan (119 mg xylose/mg protein.min) on day five and carboxymethyl cellulose (35 mg glucose/mg protein.min) on day three of solid-substrate fermentation. T. reesei showed the highest activity for Sigma cell 20 (54.8 mg glucose/mg protein.min) on day 5 of solid-substrate fermentation.
The effect of different pretreatments on SSF of corn fiber by fungal processes was examined. Corn fiber was treated at 30 oC for 2 h with alkali [2% NaOH (w/w)], alkaline peroxide [2% NaOH (w/w) and 1% H2O22 (w/w)], and by steaming at 100 oC for 2 h. Mild pretreatment resulted in improved ethanol yields for brown- and soft-rot SSF, while white-rot and Spezyme CP SSFs showed no improvement in ethanol yields.
We showed that saccharification of lignocellulosic material with a wood-rot fungal process is quite feasible. Corn fiber from wet milling was best degraded to sugars using aerobic solid state fermentation with the soft-rot fungus T. reesei . However, it was shown that both the white-rot fungus P. chrysosporium and brown-rot fungus G. trabeum had the ability to produce additional consortia of hemi/cellulose degrading enzymes. It is likely that a consortium of enzymes from these fungi would be the best approach in saccharification of lignocellulose. In all cases, a subsequent anaerobic yeast process under submerged conditions is required to ferment the released sugars to ethanol.
To our knowledge, this is the first time report on production of cellulolytic enzymes from wet-milled corn fiber using white- and brown-rot fungi for sequential fermentation of corn fiber hydrolyzate to ethanol.
Shrestha, Prachand, "Enhanced bioprocessing of lignocellulose: Wood-rot fungal saccharification and fermentation of corn fiber to ethanol" (2008). Graduate Theses and Dissertations. 11492.