Effects of Heating Treatment on the Activity and Conformation of Soybean Lipoxygenase
In this research,the effects of heating treatment conditions on LOX activity and conformation were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),circular dichroism spectroscopy,surface hy-drophobicity,fluorescence spectroscopy and molecular docking.The results showed that 120 ℃ 80 s+140 ℃ 15 s and 100 ℃ 10 min heating treatment could inhibit 67.47％and 60.53％of LOX activity.SDS-PAGE results showed that the molecular mass of the enzyme did not change significantly with heating conditions,but the band strength decreased.Moreover,heating treatment caused the decrease in α-helix and β-sheet,and increased in fluorescence intensity and surface hydrophobicity,suggesting that the greatly changed secondary and tertiary structures of LOX lead to a rapid de-cline in LOX activity.Molecular docking determined that the microenvironment surrounding the non heme iron at the ac-tive center of LOX was susceptible to temperature changes.Therefore,it was speculated that high-temperature heating treatment may affect the LOX activity by destroying the non-covalent interaction force that maintains the structure of en-zyme.High-temperature short-time elevated-temperature inactivation of LOX is an appropriate heating method.The find-ings could provide a theoretical reference for the widespread application of LOX activity control in soybean processing.
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