title: Structure and antimicrobial activity relationship of royalisin, an antimicrobial peptide from royal jelly of Apis mellifera.
abstract: Royalisin is a 5.5-kDa antibacterial peptide isolated from the royal jelly of the honeybee (Apis mellifera). The antimicrobial activity of royalisin against fungi, Gram-positive and Gram-negative bacteria has been revealed. Compared with another insect antibacterial peptide, there is an extra stretch of 11 amino acid residues at the C-terminus of royalisin. In this study, a recombinant shortened form of royalisin named as royalisin-D, was constructed without the 11 amino acid residues at the C-terminal of royalisin and linked to the C-terminal of oleosin by an inteinS fragment. The recombinant protein was overexpressed in Escherichia coli, purified by artificial oil body system and subsequently released through self-splicing of inteinS induced by the changes of temperature. The antibacterial activity of royalisin-D was compared with royalisin via minimal inhibitory concentration (MIC) assay, minimal bactericidal concentration (MBC) assay, microbial adhesion to solvents (MATS) methods, and cell membrane permeability. Furthermore, the recombinant royalisin and royalisin-D have also been treated with the reducing agent of disulfide bonds, dithiothreitol (DTT), to investigate the importance of the intra-disulfide bond in royalisin. In our results, royalisin-D exhibited similar antimicrobial activity to royalisin. Royalisin and royalisin D lost their antimicrobial activities when the intra-disulfide bonds were reduced by DDT. The intra-disulfide bond plays a more important role than the extra stretch of 11 amino acid residues at the C-terminus of royalisin in terms of the antimicrobial properties of the native royalisin.
authors.affiliation: Institute of Forest Ecology, Slovak Academy of Sciences, Zvolen, Detasched workplace Department of Molecular Apidology, Bratislava, Slovakia.
authors.forename: Katarina
authors.initials: K
authors.lastname: Bílikova
authors.affiliation: Department of Biotechnology, National Formosa University, Yunlin, Taiwan.
authors.forename: Sheng-Chang
authors.initials: SC
authors.lastname: Huang
authors.affiliation: Department of Biotechnology, National Formosa University, Yunlin, Taiwan; Department of Research and Development, Challenge Bioproducts Co., Ltd., Yunlin, Taiwan.
authors.forename: I-Ping
authors.initials: IP
authors.lastname: Lin
authors.affiliation: Institute of Forest Ecology, Slovak Academy of Sciences, Zvolen, Detasched workplace Department of Molecular Apidology, Bratislava, Slovakia.
authors.forename: Jozef
authors.initials: J
authors.lastname: Šimuth
authors.affiliation: Department of Biotechnology, National Formosa University, Yunlin, Taiwan. Electronic address: bocky@nfu.edu.tw.
authors.forename: Chi-Chung
authors.initials: CC
authors.lastname: Peng
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001516:Bees; D005227:Fatty Acids / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D019476:Insect Proteins / Q000737:chemistry / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins; D008826:Microbial Sensitivity Tests; D015394:Molecular Structure; D010539:Permeability; D011506:Proteins / Q000737:chemistry / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000378:metabolism; D057789:Staphylococcus intermedius / Q000187:drug effects / Q000378:metabolism; D000099219:Royal Jelly
chemical_list: D000900:Anti-Bacterial Agents; D005227:Fatty Acids; D019476:Insect Proteins; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; D000099219:Royal Jelly; C064722:royalisin
keywords: Antimicrobial properties; Cell membrane permeability; Royalisin; Shortened form Royalisin