The antibacterial activity of royalisin, an antimicrobial peptide from the royal jelly produced by honeybees, has been addressed extensively. However, its mechanism of action remains unclear. In this study, a recombinant royalisin, RAcc-royalisin from the royal jelly of Asian honeybee Apis cerana cerana, was expressed by fusing with glutathione S-transferase (GST) in Escherichia coli BL21, isolated and purified. The agar dilution assays with inhibition zone showed that RAcc-royalisin, similar to nisin, inhibits the growth of Gram-positive bacteria. The antibacterial activity of RAcc-royalisin was associated with its concentration, and was weakened by heat treatment ranging from 55°C to 85°C for 15 min. Both RAcc-royalisin and nisin exhibited the minimum inhibitory concentrations (MIC) of 62.5 µg/ml, 125 µg/ml, and 250 µg/ml against Gram-positive bacterial strains, Bacillus subtilis and Micrococcus flavus and Staphyloccocus aureus in the microplate assay, respectively. However, RAcc-royalisin did not show antimicrobial activity against tested Gram-negative bacterial and fungal strains. The antibacterial activity of RAcc-royalisin agrees well with the decrease in bacterial cell hydrophobicity, the leakage of 260-nm absorbing materials, and the observation by transmission electron microscopy, all indicating that RAcc-royalisin induced the disruption and dysfunction of cell walls and membranes. This is the first report detailing the antibacterial mechanism of royalisin against Gram-positive bacteria, and provides insight into the application of recombinant royalisin in food and pharmaceutical industries as an antimicrobial agent.
Major royal jelly protein 1 (MRJP1), designated apalbumin 1, has been regarded as a freshness marker of royal jelly (RJ). A MRJP1-specific peptide (IKEALPHVPIFD) identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody (anti-SP-MRJP1 antibody). Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1 (anti-R-MRJP1 antibody) reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay (ELISA) using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ (0 d). Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage (P<0.0001). Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.
We report structural features and multifunctional properties of nanocomposites based on poly(butylene terephthalate) (PBT) and exfoliated graphite (EG), which are manufactured by melt-compounding technique. Scanning electron microscope and X-ray diffraction data exhibit that graphene platelets of EG are well dispersed and exfoliated in the PBT matrix even at high EG content of 7.0 wt %. Raman spectra support that graphene platelets of EG are interacted specifically with phenyl rings of PBT chains at nanocomposite interfaces. DSC cooling and following heating thermograms of the nanocomposites demonstrate that graphene platelets of EG play a role as effective nucleating agents for PBT a-phase crystals and thus lead to accelerating the overall crystallization of the nanocomposites. Thermal stability of PBT/EG nanocomposites is improved substantially due to the gas barrier effect of graphene platelets of EG dispersed in the PBT matrix, especially at the active oxygen gas condition. Dynamic mechanical modulus of the nanocomposites is also enhanced significantly with increasing the EG content. The electrical volume resistivity of the nanocomposites is decreased dramatically from $ 10 18 to $ 10 6 X cm by forming the electrical conduction path at a certain EG content between 3 and 5 wt %. V C 2012 Wiley Periodicals, Inc. J Appl Polym Sci 125: E532-E540, 2012
To improve the physical properties of poly(trimethylene terephthalate) (PTT), a series of nanocomposites based on PTT and exfoliated graphite (EG) are prepared via melt compounding and their structures, thermal stabilities, mechanical, and electrical properties are studied. XRD and SEM show that graphene nanosheets are well dispersed in the PTT matrix without forming crystalline aggregates even at high EG content. Thermal stability and dynamic mechanical moduli of the nanocomposites are substantially improved by EG addition, and a pronounced increase in electrical volume resistivity from an insulator to almost a semiconductor is observed with increasing EG content. The electrical percolation threshold of the nanocomposites is found to be formed at the EG concentration between 3.0 and 5.0 wt.‐%.magnified image
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