Isolation, purification, and identification of protein associated with corn fiber gum.
Corn fiber gum (CFG), an alkaline hydrogen peroxide extract of the corn kernel milling byproduct "corn fiber", is a proteinaceous arabinoxylan with protein content ranging from ca. 2 to 9% by weight for CFG samples isolated from different corn milling fiber sources. Several studies have suggested that protein associated with CFG could be partly responsible for its excellent emulsifying properties in oil-in-water emulsion systems. Nevertheless, the composition and identity of the protein component has never been determined. In the present study, CFG was deglycosylated by treating with trifluoromethanesulfonic acid, and the resulting proteins were purified by passage through C18 solid phase extraction cartridges. The proteins were then separated and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein band from the gel was treated with a proteolytic enzyme, chymotrypsin, and the resulting peptides were cleaned using C18 Zip Tip pipet tips and analyzed using matrix-assisted laser desorption/ionization with automated tandem time-of-flight mass spectrometry. The partial sequences derived from the mass spectrometry analyses of the resulting chymotryptic peptides were found to be similar to the 22-kDa alpha-zein Z1 (az22z1) protein (a major storage protein in corn endosperm) when queried against the primary sequences from the National Center for Biotechnology Information database. This is the first report that this hydrophobic protein is associated with CFG and may explain why CFG is an excellent emulsifier for oil-in-water emulsion systems.
Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, 600 East Mermaid Lane, Wyndmoor, Pennsylvania 19038, United States. madhav.yadav@ars.usda.gov
Madhav P
MP
Yadav
Alberto
A
Nuñez
Kevin B
KB
Hicks
D000595:Amino Acid Sequence; D002918:Chymotrypsin / Q000378:metabolism; D045703:Emulsifying Agents / Q000737:chemistry; D006031:Glycosylation; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D053149:Plant Gums / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D012639:Seeds / Q000737:chemistry*; D003313:Zea mays / Q000737:chemistry*; D015028:Zein
D045703:Emulsifying Agents; D053149:Plant Gums; D010940:Plant Proteins; D015028:Zein; D002918:Chymotrypsin