3:00 PM - 3:15 PM
[3Hp-04] Effect of Recombinant Yeast Overexpression of Wheat LOXⅢ on the Composition Changes of Wheat Flour Dough Glutenin Subunits
Keywords:Lipoxygenase, Glutenin subunit, Solubility, Insolubility
【Objective】
Mechanistic analysis of changes in the composition of high and low molecular weight subunits of Soluble and insoluble glutenin when reacted with wheat flour, using wheat LOXⅢ isozyme overexpressed in recombinant yeast, and identification and structural analysis of LOX-sensitive polypeptides.
【Methods】
Recombinant wheat LOXⅢ isozyme (rLOXⅢ) overexpressed in recombinant yeast was purified. Soluble and insoluble glutenin fractions were prepared when purified rLOXⅢ was reacted with wheat flour, and polypeptides constituting high and low molecular weight subunits were fractionated by two-dimensional electrophoresis and RP-HPLC. LOX-sensitive polypeptides were identified by comparison with the rLOX-free control group.
【Results】
Wheat LOX isozyme III was overexpressed in yeast.
When LOX isozyme III was reacted with wheat flour, a decrease in insoluble glutenin protein and an increase in soluble glutenin protein were observed compared to the control group. At this time, an increase in high molecular weight subunit proteins constituting insoluble glutenin and a decrease in low molecular weight subunit proteins were observed, while the opposite trend was observed in soluble glutenin. The high molecular weight subunit fraction of insoluble glutenin suggested a polypeptide around 90 kDa. RP-HPLC analysis revealed differences in hydrophobic polypeptides constituting high molecular weight subunits in insoluble glutenin, particularly when reacted with LOXⅢ. Additionally, a decrease in hydrophobic polypeptides constituting low molecular weight subunits was observed.
Mechanistic analysis of changes in the composition of high and low molecular weight subunits of Soluble and insoluble glutenin when reacted with wheat flour, using wheat LOXⅢ isozyme overexpressed in recombinant yeast, and identification and structural analysis of LOX-sensitive polypeptides.
【Methods】
Recombinant wheat LOXⅢ isozyme (rLOXⅢ) overexpressed in recombinant yeast was purified. Soluble and insoluble glutenin fractions were prepared when purified rLOXⅢ was reacted with wheat flour, and polypeptides constituting high and low molecular weight subunits were fractionated by two-dimensional electrophoresis and RP-HPLC. LOX-sensitive polypeptides were identified by comparison with the rLOX-free control group.
【Results】
Wheat LOX isozyme III was overexpressed in yeast.
When LOX isozyme III was reacted with wheat flour, a decrease in insoluble glutenin protein and an increase in soluble glutenin protein were observed compared to the control group. At this time, an increase in high molecular weight subunit proteins constituting insoluble glutenin and a decrease in low molecular weight subunit proteins were observed, while the opposite trend was observed in soluble glutenin. The high molecular weight subunit fraction of insoluble glutenin suggested a polypeptide around 90 kDa. RP-HPLC analysis revealed differences in hydrophobic polypeptides constituting high molecular weight subunits in insoluble glutenin, particularly when reacted with LOXⅢ. Additionally, a decrease in hydrophobic polypeptides constituting low molecular weight subunits was observed.