Cloning and optimization of a nisin biosynthesis pathway for bacteriocin harvest

Wentao Kong, Ting Lu

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)439-445
Number of pages7
JournalACS synthetic biology
Volume3
Issue number7
DOIs
StatePublished - Jul 18 2014

Keywords

  • lactic acid bacteria
  • nisin biosynthesis
  • optimization
  • pathway engineering

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (miscellaneous)
  • Biomedical Engineering

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