Cloning and optimization of a nisin biosynthesis pathway for bacteriocin harvest.
Nisin is an important antimicrobial peptide that has enormous applications in biotechnology. Despite many encouraging efforts, its overproduction has been a long-standing challenge due to the complexity of the underlying pathway and the difficulty in genetic modification of lactic acid bacteria. Here, we cloned an entire nisin biosynthesis pathway from a nisin-producing strain (Lactococcus lactis K29) into a plasmid and transplanted the plasmid into a nisin deficient strain Lactococcus lactis MG1363, resulting in successful heterologous expression of bioactive recombinant nisin. To increase nisin harvest, we also overexpressed nisA, a gene responsible for nisin precursor production, with a set of constitutive promoters. To further optimize nisin yield, we minimized the metabolic cost of the engineered strains by integrating nisA overexpression cassettes and the recombinant pathway into a single circuit. With our rational construction and optimization, our engineered optimized strain is able to produce bioactive nisin with a yield of 1098 IU/mL, which is more than six times higher than that of the original strain.
Department of Bioengineering and Institute for Genomic Biology, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
Wentao
W
Kong
Ting
T
Lu
D001430:Bacteriocins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism; D009561:Nisin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D053038:Quorum Sensing / Q000187:drug effects; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
D001430:Bacteriocins; D011994:Recombinant Proteins; D009561:Nisin; C102526:nisin A