et al., 2011a). Upon decanting of whole stillage, emulsified oil andoil in oleosomes will partition into thin stillage, whereas, oil inunbroken matrices and that adhering to larger particles partitionwith wet cake. Non-starch carbohydrate hydrolysis enzymes, likeBluZy-P XL, can hydrolyze cell walls of unbroken cells and releaseoil from them. Protease can hydrolyze protein particles and freethe attached oil droplets as well as act on stabilizing proteins likeoleosins in thin stillage emulsion. They all contribute to increaseoil partitioning into thin stillage after decanting, and most of oil isrecovered as free oil.3.2.4. Distillers’ wet grain after enzyme treatmentsThe wet distillers grains from enzyme treatments were driedafter centrifugation and distribution of non-starch carbohydratescomponents were determined as weight % of dried samples. Sinceyields of wet distillers’ grains at the end of enzyme treatmentsdiffered based on extent of hydrolysis, we expressed the con-stituent compositions (cellulose, hemicellulose, lignin) as % ofstarting 100 g dry corn. Hydrolysis of non-starch carbohydrateby BluZy-P XL resulted in lower amount of fiber content (bothNDF and ADF), except for Treatment A, which had higher valuesthan control group (Table 3). Control and Treatment A had signifi-cantly higher amount of fiber components than other treatments,whereas, protease/phytase treatment (Treatment D) had lowest.Treatments B and C were better stages than A for BluZy-P XL treat-ment because lower amount of non-digestible carbohydrates werefound in wet grains. Additional protease/phytase addition duringfermentation decreased that even further. It may be explainedby the further structural decomposition leading to improvementin enzymes activity. Protease hydrolyzes protein and expose cellwall carbohydrates to other enzymes like BluZy-P XL. Moreover,phytase can hydrolyze phytic acid, which can chelate with metaland may reduce or inhibit enzyme activity, especially chelationof calcium, which could be detrimental to glycoamylase-type ofenzymes. Decreasing phytic acid content in corn slurry can alsoimprove activity of carbohydrate hydrolysis enzymes (Mikulskiet al., 2014). Cell solubles, which is the most digestible partin DDGS, also decreased with enzyme treatments. BluZy-P XLand protease/phytase treatments degraded larger molecules likepolysaccharides, proteins; partitioning of these lower molecularweight molecules occurred in thin stillage fraction after decanting.Cell solubles, including protein, lipid, sugars and starch may alsohave been released from wet grains during enzyme hydrolysis andmoved to thin stillage. The solid mass distribution in thin stillageand wet cake (Table 2) supports this explanation.4. ConclusionThis study demonstrated that incorporation of hydrolyticenzymes, including non-starch hydrolase, protease, and phytase,at their optimized conditions and process stages can promote fer-mentation performance for corn dry-grind process for ethanol. Itcan also enhance partitioning of solids and oil into liquid fraction(thin stillage) and produce DDGS with lower amounts of nondi-gestible carbohydrates. Best process performance was obtainedwith 1.5 h incubation with BluZy-P XL after liquefaction (50◦C) andprotease/phytase addition during fermentation. This also resultedin favorably modified distillers grains that could be envisioned tohave uses in monogastric feeding.AcknowledgementsWe thank Direvo Inc., and LincolnWay Energy for providing withhydrolytic enzyme mix and other fermentation additives for thisstudy. We also appreciate the Department of Food Engineering andBioprocess Technology, Asian Institute of Technology for facilitat-ing first author’s stay at Iowa State University.