Characterization of Lipoxygenases from Soybean Leaves and Stems

Location

CSU 253/254/255

Start Date

12-4-2004 1:45 PM

End Date

12-4-2004 3:15 PM

Student's Major

Chemistry and Geology

Student's College

Science, Engineering and Technology

Mentor's Name

Theresa Salerno

Mentor's Department

Chemistry and Geology

Mentor's College

Science, Engineering and Technology

Description

Lipoxygenases (LOX) are biological enzymes collectively called dioxygenases and they catalyze the peroxidation (addition of molecular oxygen) of fatty acids. LOX enzymes use linolenic acid and linoleic acids as their substrates. The characterization of soybean seed LOX enzymes has been extensively studied, but few studies on the Leaf LOX enzymes have been done. The purpose of this study is to characterize and compare LOX enzymes from leaves and stems from soybeans and specifically to determine if seed LOX-free strains have different leaf and stem LOX enzymes. The characterization will include quantitative and qualitative analysis of these enzymes. These enzymes will be identified based on differences in their isoelectric point (pi) values. Five strains of soybean plants were grown for 7 days after germination. On the seventh day, stems and leaves were harvested and the LOX enzymes were extracted, and separated by charge using Isoelectric Focusing gels. Specific activity staining was done in the presence of linoleic acid as the substrate.. The LOX enzymes were then quantitated by densitometric analysis. Preliminary results show that there are more LOX enzymes in the leaves than in the stems. There are higher relative amounts of LOX enzymes with lower pi values in the leaves. Seed LOX-free strains still have several leaf and stem LOX enzymes. Not all strains show identical LOX enzyme patterns.

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Apr 12th, 1:45 PM Apr 12th, 3:15 PM

Characterization of Lipoxygenases from Soybean Leaves and Stems

CSU 253/254/255

Lipoxygenases (LOX) are biological enzymes collectively called dioxygenases and they catalyze the peroxidation (addition of molecular oxygen) of fatty acids. LOX enzymes use linolenic acid and linoleic acids as their substrates. The characterization of soybean seed LOX enzymes has been extensively studied, but few studies on the Leaf LOX enzymes have been done. The purpose of this study is to characterize and compare LOX enzymes from leaves and stems from soybeans and specifically to determine if seed LOX-free strains have different leaf and stem LOX enzymes. The characterization will include quantitative and qualitative analysis of these enzymes. These enzymes will be identified based on differences in their isoelectric point (pi) values. Five strains of soybean plants were grown for 7 days after germination. On the seventh day, stems and leaves were harvested and the LOX enzymes were extracted, and separated by charge using Isoelectric Focusing gels. Specific activity staining was done in the presence of linoleic acid as the substrate.. The LOX enzymes were then quantitated by densitometric analysis. Preliminary results show that there are more LOX enzymes in the leaves than in the stems. There are higher relative amounts of LOX enzymes with lower pi values in the leaves. Seed LOX-free strains still have several leaf and stem LOX enzymes. Not all strains show identical LOX enzyme patterns.