Thermal Instability Induced by 4-Hydroxy-2-Nonenal in Beef Myoglobin

Abstract

The secondary products of lipid oxidation, such as 4-hydroxy-2-nonenal (HNE), compromise myoglobin (Mb) redox stability and can thus impact thermal stability. Previous studies examined HNE-induced redox instability in beef Mb, whereas investigations are yet to be undertaken to evaluate the relationship between lipid oxidation and thermal stability of beef Mb. Therefore, the objective of the present study was to investigate the direct influence of HNE on thermal stability of beef Mb at meat conditions. Beef oxymyoglobin (0.15 mM) was incubated with HNE (1.0 mM) at pH 5.6 and 4°C for 21 d in the dark. Metmyoglobin formation, percentage Mb denaturation (PMD), and HNE adduction sites in Mb were examined on days 0, 7, 14, and 21. The experiment was replicated 3 times (n = 3). The data were evaluated using the MIXED procedure of SAS, and the differences among means were detected at the 5% level using the least significant difference test. The HNE-treated samples exhibited greater (P < 0.05) metmyoglobin formation and PMD than the controls. Additionally, the PMD difference between HNE-treated and control samples increased (P < 0.05) over time. Mass spectrometric analyses indicated that the number of HNE adduction sites increased with storage, and 6 histidines (positions 24, 36, 64, 93, 113, and 152) were adducted on day 21. HNE adduction at the distal histidine (position 64), which is critical to he me stability, was observed only on days 14 and 21. An increase in PMD on days 14 and 21 in HNE-treated samples could be partially due to the adduction at distal histidine. These findings indicated that HNE compromises thermal stability of beef Mb, possibly through altering the conformation of the heme protein by nucleophilic adduction.

Keywords

• beef myoglobin
• redox stability
• mass spectrometry
• thermal stability
• lipid oxidation