Nisin-controlled extracellular production of apidaecin in <Emphasis Type="Italic">Lactococcus lactis</Emphasis> |
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Authors: | Xu-xia Zhou Yan-bo Wang Yuan-jiang Pan Wei-fen Li |
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Institution: | (1) Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Zhejiang University Road 38#, Hangzhou, China;(2) Microbiology Division, the Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Zhejiang University, Hangzhou, 310029, China;(3) Food Quality and Safety Department, Zhejiang Gongshang University, Hangzhou, China |
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Abstract: | 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. |
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Keywords: | Extracellular production Apidaecin Purification Antibacterial activity Cytotoxicity |
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