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Nisin-selectable food-grade secretion vector for Lactococcus lactis.

Ruiqing Li1, Timo M Takala, Mingqiang Qiao

  • 1College of Life Sciences, Nankai University, Tianjin, China.

Biotechnology Letters
|December 25, 2010
PubMed
Summary

A novel nisin-resistance gene (nsr) enabled the creation of a food-grade secretion plasmid, pLEB690, for Lactococcus lactis. This vector facilitates the expression and secretion of pediocin AcH, offering potential applications in the food industry.

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Area of Science:

  • Food Microbiology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Nisin resistance in Lactococcus lactis is crucial for industrial applications.
  • Development of efficient food-grade secretion vectors is essential for producing antimicrobial peptides.
  • Identification of novel genes conferring nisin resistance can lead to improved genetic tools.

Purpose of the Study:

  • To construct and characterize a novel food-grade secretion plasmid in Lactococcus lactis.
  • To utilize a nisin-resistance gene for developing a selectable marker in a food-grade vector.
  • To demonstrate the functionality of the developed vector for secreting pediocin AcH.

Main Methods:

  • Isolation of a nisin-resistant Lactococcus lactis strain (TML01).

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  • Identification of a nisin-resistance gene (nsr) with high homology to known nsr genes.
  • Construction of a food-grade secretion plasmid (pLEB690) using the identified nsr gene.
  • Expression and secretion of the pediocin AcH gene (papA) using the pLEB690 vector in L. lactis.

    • Main Results:

    • A Lactococcus lactis strain TML01 exhibiting nisin resistance was isolated.
    • A novel nsr gene was identified with 99% homology to existing nsr genes.
    • The constructed pLEB690 plasmid demonstrated functionality as a secretion vector, enabling selection with up to 5 μg nisin/ml.
    • Successful expression and secretion of pediocin AcH were achieved using the pLEB690 vector.

    Conclusions:

    • A functional food-grade secretion vector, pLEB690, was successfully constructed based on a novel nisin-resistance gene.
    • The pLEB690 vector provides a valuable tool for the food industry, facilitating the secretion of antimicrobial peptides like pediocin AcH.
    • This research contributes to the development of advanced genetic tools for food biotechnology.