Proteomic identification and enzymatic activity of buckwheat (Fagopyrum esculentum) honey based on different assays

Abstract

Buckwheat (Fagopyrum esculentum Moench) blossom honey was analysed for enzymatic activity and protein content. Proteins were separated by one-dimensional denaturing electrophoresis (1DE) in sodium dodecyl sulphate polyacry-lamide gel (SDS-PAGE) or resolved by two-dimensional denaturing gel electrophoresis (2DE). Using different mass spectrometry (MS) techniques, the main proteins of plant and honeybee Apis mellifera carnica, Pollmann origin were identified. Using direct MS analysis of honeybee proteome, a total of 87 proteins were identified in buckwheat honey. Proteins of honeybee origin included major royal jelly protein (MRJP), uncharacterized honeybee protein, high glutamic acid storage protein and enzymes α-glucosidase, α-amylase and glucosylceramidase. Separation of proteins by 1DE and 2DE revealed a smaller number of proteins, including 6 proteins of bee origin and 11 of yeast origin. In addition, catalase activity and glucose oxidase activity in buckwheat honey were estimated on the native PAGE. Our study demonstrates a useful approach to identification of honey proteins, which play an important role in human nutri-tion and immune defence system.