Formation of Protein-Bound Maillard Reaction Products during the Storage of Manuka Honey.

Honey from the nectar of the Manuka tree (Leptospermum scoparium) grown in New Zealand contains high amounts of antibacterial methylglyoxal (MGO). MGO can react with proteins to form peptide-bound Maillard reaction products (MRPs) such as Nε-carboxyethyllysine (CEL) and "methylglyoxal-derived hydroimidazolone 1" (MG-H1). To study the reactions of MGO with honey proteins during storage, three manuka honeys with varying amounts of MGO and a kanuka honey (Kunzea ericoides) spiked with various MGO concentrations up to 700 mg/kg have been stored at 37 °C for 10 weeks, and the formation of protein-bound MRPs has been analyzed via high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) following isolation of the protein fraction and enzymatic hydrolysis. During storage, contents of protein-bound CEL and MG-H1 increased continuously, directly depending on the MGO content. For honeys with large amounts of MGO, a slower formation of Nε-fructosyllysine (FL) was observed, indicating competing reactions of glucose and MGO with lysine. Furthermore, the lysine modification increased with storage independently from the MGO concentration. Up to 58-61% of the observed lysine modification was explainable with the formation of CEL and FL, indicating that other reactions, most likely the formation of Heyns products from lysine and fructose, may play an important role. Our results can contribute to the authentication of manuka honey.

3-Phenyllactic Acid and Polyphenols Are Substances Enhancing the Antibacterial Effect of Methylglyoxal in Manuka Honey.

Manuka honey is known for its unique antibacterial activity, which is due to methylglyoxal (MGO). After establishing a suitable assay for measuring the bacteriostatic effect in a liquid culture with a time dependent and continuous measurement of the optical density, we were able to show that honey differs in its growth retardingeffect on Bacillus subtilis despite the same content of MGO, indicating the presence of potentially synergistic compounds. In model studies using artificial honey with varying amounts of MGO and 3-phenyllactic acid (3-PLA), it was shown that 3-PLA in concentrations above 500 mg/kg enhances the bacteriostatic effect of the model honeys containing 250 mg/kg MGO or more. It has been shown that the effect correlates with the contents of 3-PLA and polyphenols in commercial manuka honey samples. Additionally, yet unknown substances further enhance the antibacterial effect of MGO in manuka honey. The results contribute to the understanding of the antibacterial effect of MGO in honey.