title: Expression of Acc-Royalisin gene from royal jelly of Chinese honeybee in Escherichia coli and its antibacterial activity.
abstract: Royalisin is an antibacterial peptide found in Royal Jelly. Two gene fragments of Chinese honeybee (Apis cerana cerana) head, 280 bp cDNA encoding pre-pro-Acc-royalisin (PPAR) of 95 amino acid residues, and 165 bp cDNA encoding mature Acc-royalisin (MAR) of 51 amino acid residues were cloned into the pGEX-4T-2 vector. They were then transformed individually into Escherichia coli for expression. Two expressed fusion proteins, glutathione S-transferase (GST)-PPAR of 36 kDa and GST-MAR of 32 kDa were obtained, which were cross reacted with GST antibody accounting for up to 16.3% and 15.4% of bacterial protein, respectively. In addition, 41% of GST-PPAR and nearly 100% of GST-MAR were soluble proteins. Both lysates of the two purified fusion proteins displayed antibacterial activities, similar to that of nisin against Gram-positive bacteria strains, Staphylococcus aureus, Bacillus subtilis and Micrococcus luteus. MAR peptide released from the thrombin-cleaved GST-MAR fusion protein has a stronger antibacterial activity than that of GST-MAR fusion protein.
authors.affiliation: Department of Food Science and Nutrition, Zhejiang University, 268 Kaixuan Road, Hangzhou, Zhejiang, China. shenlirong@zju.edu.cn
authors.forename: Lirong
authors.initials: L
authors.lastname: Shen
authors.forename: Meihui
authors.initials: M
authors.lastname: Ding
authors.forename: Liwen
authors.initials: L
authors.lastname: Zhang
authors.forename: Feng
authors.initials: F
authors.lastname: Jin
authors.forename: Weiguang
authors.initials: W
authors.lastname: Zhang
authors.forename: Duo
authors.initials: D
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001483:Base Sequence; D001516:Bees / Q000235:genetics / Q000502:physiology*; D002681:China; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D020224:Expressed Sequence Tags; D005227:Fatty Acids / Q000235:genetics*; D005786:Gene Expression Regulation; D006094:Gram-Positive Bacteria / Q000187:drug effects; D006722:Honey; D007301:Insect Hormones; D036341:Intercellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D000099219:Royal Jelly
chemical_list: D000900:Anti-Bacterial Agents; D004247:DNA; D017931:DNA Primers; D005227:Fatty Acids; D007301:Insect Hormones; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D000099219:Royal Jelly; C064722:royalisin