Nisin-controlled extracellular production of apidaecin in Lactococcus lactis.
Apidaecins are heat-stable, nonhelical antibacterial peptides isolated from lymph fluid of the honeybee (Apis mellifera). These peptides are active against a wide range of gram-negative bacteria and they are the most prominent components of the honeybee humoral defense against microbial invasion. In the present study, one isoform of apidaecin, apidaecin Ho, was expressed extracellularly in the food-grade bacterium Lactococcus lactis. Results showed that expression driven by the lactococcal nisA promoter and Usp45 signal peptide resulted in efficient secretion of apidaecin in L. lactis subsp. cremoris NZ9000. Recombinant apidaecin was purified by gel filtration and semipreparative RP-HPLC, and about 10 mg active recombinant apidaecin was obtained from 1,000 ml culture. This is the first report on the nisin-controlled extracellular production of active apidaecin in L. lacits. The expression and delivery of apidaecin in the food-grade L. lactis may provide a clue to facilitate the widespread application of apidaecin in the control and prevention of gram-negative bacteria infections of human and animals.
Zhou
Xu-Xia
XX
Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou, China.
Wang
Yan-bo
YB
Pan
Yuan-jiang
YJ
Li
Wei-fen
WF
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D001516:Bees / Q000737:chemistry; D018938:Caco-2 Cells; D004926:Escherichia coli / Q000187:drug effects; D005818:Genetic Engineering; D005822:Genetic Vectors; D006801:Humans; D013294:Lactococcus lactis / Q000235:genetics* / Q000378:metabolism*; D013058:Mass Spectrometry; D009561:Nisin / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D011994:Recombinant Proteins; C061361:apidaecin; D009561:Nisin