Steven R. Fox

Comparison of laboratory and pilot-plant corn wet-milling procedures

ABSTRACT One waxy and three regular yellow dent corn hybrids were wet milled by using two scales of laboratory procedures (modified 100-g and 1-kg) and a pilot-plant procedure (10-kg). The modified 100-g and 1-kg laboratory procedures gave similar yields of wet-milling fractions. Starch yields and recoveries were significantly lower for the pilot-plant procedure, whereas gluten and fiber yields were greater because of their high contents of unrecovered starch. Protein contents of the starches obtained by all three procedures were within commercially acceptable limits (<0.50% db for normal dent corn and <0.30% for waxy corn). Rankings for starch yields and starch recoveries for the four hybrids, having very different physical and compositional properties, were the same for all three procedures. The harder the grain, the lower the yield and recovery of starch. Least significant differences (P < 0.05) for starch yield were 0.8% for the modified 100-g procedure, 1.2% for the 1-kg procedure, and 2.0% for the p...

A 10-g Laboratory Wet-Milling Procedure for Maize and Comparison With Larger Scale Laboratory Procedures

ABSTRACT A very small scale laboratory procedure (≈10 g) is needed to test wet-milling characteristics of corn when amounts of corn available for testing are quite limited. The objective of this study was to downscale 100-g laboratory wet-milling methods already widely used to measure wet-milling properties of 10 g of corn. A Standard 100-g procedure, a Modified 100-g procedure, and an Experimental 10-g procedure were compared using three corn hybrids with known differences in wet-milling properties. All three procedures ranked most fraction yields (all except for germ) of the three hybrids the same. Germ separation was conducted differently for each procedure and probably accounts for these differences. Flotation and screening methods were likely affected by germ density and germ size, and hand-picking the germ was efficient in recovering a pure germ fraction. The two 100-g procedures were performed very similarly except for fiber recovery. The Modified 100-g procedure was more efficient in recovering fi...

Fractionation of transgenic corn seed by dry and wet milling to recover recombinant collagen‐related proteins

Corn continues to be considered an attractive transgenic host for producing recombinant therapeutic and industrial proteins because of its potential for producing recombinant proteins at large volume and low cost as coproducts of corn seed‐based biorefining. Efforts to reduce production costs have been primarily devoted to increasing accumulation level, optimizing protein extraction conditions, and simplifying the purification. In the present work, we evaluated two grain fractionation methods, dry milling and wet milling, to enrich two recombinant collagen‐related proteins; thereby, reducing the amount and type of corn‐derived impurities in subsequent protein extraction and purification steps. The two proteins were a full‐length human recombinant collagen type I alpha 1(rCIα1) chain with telopeptides and peptide foldon to effect triple helix formation and a 44‐kDa rCIα1 fragment. For each, ∼60% of the rCIα1s in the seed was recovered in the dry‐milled germ‐rich fractions making up ca. 25% of the total kernel mass. For wet milling, ∼60% of each was recovered in three fractions accounting for 20–25% of the total kernel mass. The rCIα1s in the dry‐milled germ‐rich fractions were enriched three to six times compared with the whole corn kernel, whereas the rCIα1s were enriched 4–10 times in selected wet‐milled fractions. The recovered starch from wet milling was almost free of rCIα1. Therefore, it was possible to generate rCIα1‐enriched fractions by both dry and wet milling along with rCIα1‐free starch using wet milling. Because of its simplicity, the dry milling procedure could be accomplished on‐farm thus minimizing the risk of inadvertent release of viable transgenic seeds.

Recovering corn germ enriched in recombinant protein by wet-fractionation

Corn wet-fractionation processes (quick-germ fractionation and traditional wet milling) were evaluated as means of recovering fractions rich in recombinant collagen-related proteins that were targeted for expression in the germ (embryo) of transgenic corn. Transgenic corn lines accumulating a recombinant full-length human collagen type-I-alpha-1 (full-length rCIalpha1) or a 44-kDa rCIalpha1 fragment targeted for seed expression with an embryo-specific promoter were used. Factors to consider in efficient recovery processes are the distribution of the peptides among botanical parts and process recovery efficiency. Both recombinant proteins were distributed 62-64% in germ comprising about 8.6% of the dry grain mass; 34-38% in the endosperm comprising 84% of the dry grain mass; 1.7% in the pericarp comprising about 5% of the dry mass; and 1% in the tip-cap comprising 1.5-2% of the dry mass. The quick-germ method employed a short steeping period either in water or SO(2)-lactic acid solution followed by wet-milling degermination to recover a germ-rich fraction. Of the total recombinant protein expressed in germ, the quick-germ process recovered 40-43% of the total recombinant protein within 6-8% of the corn mass. The traditional corn wet-milling process produced higher purity germ but with lower recovery (24-26%) of the recombinant protein. The two quick-germ methods, using water alone or SO(2)-lactic acid steeping, did not substantially differ in rCIalpha1 recovery, and the quick-germ processes recovered germ with less leaching and proteolytic losses of the recombinant proteins than did traditional wet milling. Thus, grain fractionation enriched the recombinant proteins 6-fold higher than that of unfractionated kernels. Such enrichment may improve downstream processing efficiency and enable utilizing the protein-lean co-products to produce biofuels and biorenewable chemicals by fermenting the remaining starch-rich fractions.

Wet-Milling Characteristics of 10 Lines from Germplasm Enhancement of Maize Project Compared with Five Corn Belt Lines

ABSTRACT The use of corn (Zea mays L.) hybrids with high grain yield and starch extractability has steadily increased in the processing industry. In light of widespread corn seed industry participation in the Germplasm Enhancement of Maize Project (GEM), which seeks to enhance exotic germplasm, future hybrids may contain more exotic sources in genetic backgrounds. It is necessary to establish and monitor physical, compositional, and milling characteristics of the new exotic breeding materials to determine the processing value. The present study was conducted to determine the wet-milling characteristics of a set of GEM lines compared with typical Corn Belt lines. Ten GEM lines introgressed with exotic materials from Argentina, Chile, Cuba, Florida, and Uruguay and previously identified as having different starch yields, three commercial inbred lines, and two public inbred lines (B73 and Mo17) were analyzed using both near-infrared transmittance (NIT) and a 100-g wet-milling procedure. There were statistica...

Variation Among Physical, Compositional, and Wet-Milling Characteristics of the F1 Generation of Corn Hybrids of Introgressed Exotic and Adapted Inbred Lines

ABSTRACT Hybrids with high grain yield and higher starch, protein, or oil content are available to corn growers; however, they result from crossing adapted Corn Belt inbred lines that rarely include exotic germplasm. This study was conducted to determine whether Corn Belt lines introgressed with exotic materials from Argentina, Chile, Uruguay, Cuba, and Florida have appropriate wet-milling characteristics in their hybrids. Ten lines from the Germplasm Enhancement of Maize (GEM) project with different starch contents were crossed to three adapted inbred lines used as testers. The B73×Mo17 hybrid was used as a control. The F1 generation of these 30 experimental hybrids was analyzed using both near-infrared transmittance (NIT) technology and a 100-g modified wet-milling procedure, and measuring test and 1,000-kernel weight. There was great variation among physical, compositional, and wet-milling characteristics of the experimental hybrids, suggesting that exotic germplasm can be used to improve wet-milling c...

Physical, Compositional, and Wet Milling Characteristics of Grain from Crosses of Corn Inbreds with Exotic and Nonexotic Background

ABSTRACTCorn breeders have developed hybrids with enhanced compositional characteristics, but exotic germplasm represents little of the germplasm base used to produce these hybrids. Effects of the exotic germplasm on physical, compositional, and wet-milling properties as well as the proximate composition of recovered fractions need to be determined before these materials are of value to the corn processing industry. Ten lines from the Germplasm Enhancement of Maize (GEM) project with exotic germplasm introgressed from Argentina, Chile, Uruguay, Cuba, and Florida were crossed to three adapted inbred lines (testers) and grain from the resulting 30 hybrids were evaluated for physical, compositional, and wet-milling characteristics and the expression of heterosis in these variables. The B73xMo17 adapted public hybrid was used as control. Grain obtained by self-pollination of the hybrid plants was analyzed using near-infrared transmittance (NIT) technology and a 100-g wet-milling procedure. There was great var...

Pilot-plant wet-milling process for producing corn gluten meal

ABSTRACT A pilot-plant wet-milling process was specially used to produce corn gluten meal (CGM) with a relatively high protein content (49.8% from Pioneer 3394 and 53.7% from Wilson D110). The protein content of the CGM obtained from a starch table in our pilot-plant procedure was similar to the results obtained by using the hydrocyclone method. Wilson D110 has a higher protein content in its corn kernels, producing higher yield and protein content gluten than the Pioneer 3394, which has lower protein content in the corn. The effects of drying method and temperature on the color of CGM were also discussed.