Expression of Acc-Royalisin gene from royal jelly of Chinese honeybee in Escherichia coli and its antibacterial activity.
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.
Shen
Lirong
L
Department of Food Science and Nutrition, Zhejiang University, 268 Kaixuan Road, Hangzhou, Zhejiang, China. shenlirong@zju.edu.cn
Ding
Meihui
M
Zhang
Liwen
L
Jin
Feng
F
Zhang
Weiguang
W
Li
Duo
D
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
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