Major royal jelly proteins as markers of authenticity and quality of honey / Glavni proteini matične mliječi kao markeri izvornosti i kakvoće meda

Abstract

Until now, the properties of honey have been defined based exclusively on the content of plant components in the nectar of given plant. We showed that apalbumin1, the major royal jelly (RJ) protein, is an authentic and regular component of honey. Apalbumin1 and other RJ proteins and peptides are responsible for the immunostimulatory properties and antibiotic activity of honey. For the quantification of apalbumin1, an enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal anti-apalbumin1 antibody. The method is suitable for honey authenticity determination; moreover it is useful for detection of the honey, honeybee pollen and RJ in products of medicine, pharmacy, cosmetics, and food industry, where presences of these honeybee products are declared. Results from the analysis for presence and amount of apalbumin1 in honeys will be used for high-throughput screening of honey samples over the world. On the basis of our experiments which show that royal jelly proteins are regular and physiologically active components of honey we propose to change the definition of honey (according to the EU Honey Directive 2001/110/EC) as follows: Honey is a natural sweet substance produced by honey bees from nectar of plants or from secretions of plants, or excretions of plant sucking insects, which honey bees collect, transform by combining with major royal jelly proteins and other specific substances of their own, deposit, dehydrate, store and leave in the honey comb to ripen and mature.

An Effective Method of Isolating Honey Proteins

“…Obtained one-dimensional protein maps (Figure 3) have sufficient resolution and quality and they do not deviate from recently published standards [6,7,8,15] and may even exceed them. We cannot explain exactly why all lanes, with proteins obtained after phenol extraction, are the most intense on gels (Figure 3).…”

“…These regulations describe sugar and 5-hydroxymethylfurfural (HMF) content, acidity, diastase activity, and moisture as the maturity indicators, and as purity control parameters: ash content, electrical conductivity and insoluble solids in water [19]. However, recently also protein content and variability are seen as another parameter for quality [15,20], origin [12,21] and authenticity [6,8]. Increasing interest in honey proteins seems to be a very important factor leading to the improvement of the honey protein analysis methodology.…”

“…The second identified protein was major royal jelly protein 1, one of the most abundant bee-origin proteins in honey. This protein is also proposed as a marker of honey authenticity and quality [8] and a few years ago even ELISA test using MRJP (Major Royal Jelly Protein) was developed for screening of honey adulteration based on the addition of industrial sugar syrups and/or of honey from bees fed with sucrose syrup [50].…”

“…The most abundant among them are major royal jelly proteins [4,5,6,7] but there are also present enzymes such as α- and β- glucosidase (invertase), α- and β- amylase (diastase), glucose oxidase, and others [3]. The proteins present in honey are widely used as markers of honey authenticity and adulteration tries [6,8] and also as a quality indicator [8,9]. Nevertheless, generally, this proteome is still very poorly researched, mostly because of the low amount of protein in honey (0.1–0.5%) [4].…”

The Effect of Diet on the Composition and Stability of Proteins Secreted by Honey Bees in Honey

“…The second group of proteins contains major royal jelly proteins 1–5 (mostly MRJP1 [10]), royalisin (known as defensin-1 [11]) and apisimin [12]. These proteins are important for honey’s antimicrobial activity and are the most abundant proteins of the total protein quantity [13]. The antimicrobial activity of MRJPs, combined with substances (e.g., H 2 O 2 , gluconic acid) generated during sugar conversion, are thought to prevent honey from microbial spoilage.…”