Kikuji Yamaguchi Principles of Natural Beekeeping: A Novel Bio-Method of Natural Beekeeping for High Quality Royal Jelly Production

Many serious problems, such as artificial control and overworking of honey bee colonies, deterioration of bee products due to incorrect treatment and inadequate environments, have been postulated in recent beekeeping which should be resolved for sustainable development of modern highly profitable beekeeping in the future. Thus, a novel beekeeping method, Kikuji Yamaguchi Method of Natural Beekeeping (KYAMENABEE), was established for the natural royal jelly (RJ) preparation and the biological and pharmacological properties were examined for evaluation of the authenticity of royal jelly products. RJ samples prepared by KYAMENABEE and ordinal beekeeping were subjected to the quantitative analyses of 10HDA and MRJP1 multimer, identification of functional substance based on the effective growth and development of queen bees, stability of the functional substance, proliferative activity of human and animal cells. The content of 10HDA and MRJP1 multimer in RJ prepared by KYAMENABEE were significantly higher than that prepared by ordinal beekeeping. The biological and pharmacological activities were also superior for RJ prepared by KYAMENABEE than that by ordinal beekeeping. Thus, it might be important to use a novel beekeeping method, KYAMENABEE, in order to produce high quality RJ for sustainable development of biopharmaceutical beekeeping

Glavni proteini matične mliječi kao markeri izvornosti i kakvoće meda

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.Do sada su svojstva meda bila definirana isključivo na temelju sadržaja komponenti nektara određene biljke. Mi smo pokazali da je apalbumin1, glavni protein matične mliječi, izvoran i uobičajeni sastojak meda. Apalbumin1, ostali proteini matične mliječi i peptidi odgovorni su za imunostimulatorna svojstva i antibiotsko djelovanje meda. Korištenjem poliklonalnog anti-apalbumin 1 protutijela osmišljen je imunoenzimski test (ELISA) za kvantifikaciju apalbumina 1. Metoda je ne samo prikladna za utvrđivanje izvornosti meda nego i korisna za detekciju meda, peluda i matične mliječi u medicinskim, farmaceutskim, kozmetičkim i prehrambenim proizvodima na kojima je naznačena prisutnost pčelinjih proizvoda. Rezultati analize prisutnosti i količine apalbumina 1 koristit će se za probir velike količine uzoraka meda diljem svijeta. Na temelju naših eksperimenata, koji pokazuju da su proteini matične mliječi uobičajene i fiziološki aktivne komponente meda, predlažemo izmjenu definicije meda (na temelju Direktive EU-a o medu 2001/110/EC): Med je prirodna slatka tvar koju pčele proizvode od nektara ili izlučevina biljaka ili izlučevina insekata koji sišu biljke. Nju pčele skupljaju, pretvaraju kombinacijom glavnih proteina matične mliječi i ostalih vlastitih specifičnih tvari, polažu, dehidriraju, pohranjuju i ostavljaju u saću da sazrije

A rapid method to isolate soluble royal jelly proteins

AuthorSoluble royal jelly (RJ) proteins (SRJPs) include the major RJ protein (MRJP) family, which contribute to the physiological actions of RJ. Although SRJPs are prepared using conventional methods involving dialysis and centrifugation, dialysis is a time-consuming process. We have therefore developed a simple method to isolate SRJPs from RJ. This new method produces 20-fold higher levels of SRJPs than that of the conventional procedure; hence, the levels obtained by the new and existing methods were compared. A 1-h ultracentrifugation separated SRJPs in the supernatant into upper, middle and lower layers. Each layer was analyzed by size-exclusion HPLC, SDS–PAGE and 2-DE. The upper and middle layers contained MRJP2 (52 kDa) and MRJP3 (60–70 kDa), while the lower layer contained MRJP1 (290 kDa). In nature, MRJP1 is a monomer and/or oligomer. When the lower layer was analyzed by Superose 12 HPLC, MRJP1 was predominantly an oligomer. Our MRJP isolation method reduces the procedure time by using ultracentrifugation without dialysis to obtain SRJPs and produces layers containing MRJP1 oligomers, MRJP2 and MRJP3