title: Application of proteomics to hordein screening in the malting process.
abstract: This study was undertaken to investigate the effect of the malting process on hordein composition. For this purpose, combination of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and the method of isotopic peptides labeling iTRAQ was used. Barley proteins are essential components determining the quality of both malt and beer. Since hordeins represent the most abundant proteins accounting for about 40-50% of total protein fraction of mature barley grain, our research was focused on them. In this respect, the proteins of interest were extracted from milled samples of barley grain, germinated barley grain (samples collected at different time intervals), green malt and malt, respectively. Particular hordein extracts were firstly fractionated via SDS- PAGE, which was used as a relatively rapid and reliable technique providing information about hordein profile of analyzed samples. Then, separated proteins were in-gel digested and resulting peptides were measured by mass spectrometry. In addition, the chosen proteins, after in-gel digestion, were subjected to the iTRAQ method and the screening of proteins during malting process was evaluated. Our results have revealed that most of the hordein components present in the barley grain can be found in all stages of the malting process as well as in the final malt. The amount of hordeins decreases during the malting process; in the case of C hordein, the protein decrease is approximately 65%. On the other hand, significant degradation of D hordein was detected. The suggested procedure can be used to follow the development of the hordein profile during germination, which is of great technological importance in beer production.
authors: Institute of Analytical Chemistry of the ASCR, Brno, Czech Republic. flodrova@iach.cz
authors: Dana
authors: D
authors: Flodrová
authors: Jiří
authors: J
authors: Ralplachta
authors: Dagmar
authors: D
authors: Benkovská
authors: Janette
authors: J
authors: Bobálová
mesh_terms: D000595:Amino Acid Sequence; D001515:Beer; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005983:Glutens / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D001467:Hordeum / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D012639:Seeds / Q000737:chemistry / Q000378:metabolism; D020539:Sequence Analysis, Protein / Q000379:methods
chemical_list: D005983:Glutens