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Construction of a shuttle expression vector for lactic acid bacteria.

Tejinder Kaur1, Praveen P Balgir2, Baljinder Kaur2

  • 1Department of Biotechnology, Punjabi University, Patiala, 147002, India. tejinder.kaur.6@gmail.com.

Journal, Genetic Engineering & Biotechnology
|November 19, 2019
PubMed
Summary

A new shuttle vector, pPBT-GFP, was developed for lactic acid bacteria (LAB) to enhance industrial strains. This vector enables the expression of pediocin, green fluorescent protein (GFP), and bile salt hydrolase (BSH) in LAB hosts.

Keywords:
Bile salt hydrolaseExpression vectorGreen fluorescent proteinHeterologous gene expressionLactic acid bacteriaPlasmid vector

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Lactic acid bacteria (LAB) are crucial in food fermentation, with genetic engineering offering potential for strain improvement.
  • Biosafety concerns necessitate well-characterized cloning and expression systems derived from LAB plasmids.
  • Developing robust expression systems is key to enhancing the industrial applications of LAB.

Purpose of the Study:

  • To develop a novel LAB/LAB shuttle expression vector, pPBT-GFP.
  • To enable efficient cloning and expression of heterologous genes in LAB.
  • To provide a tool for the genetic improvement of commercially valuable LAB strains.

Main Methods:

  • Construction of the pPBT-GFP shuttle vector utilizing characterized LAB plasmids.
  • Transformation of Pediococcus acidilactici MTCC 5101 and Lactobacillus brevis MTCC 1750.
  • Expression analysis of pediocin, green fluorescent protein (GFP), and heterologous bile salt hydrolase (BSH).

Main Results:

  • The pPBT-GFP vector demonstrated replication in both P. acidilactici and L. brevis with low copy numbers.
  • Pediocin and GFP were successfully expressed as selectable markers.
  • Efficient expression and high specific activity of heterologous BSH were observed in both host strains.

Conclusions:

  • The developed pPBT-GFP vector is capable of replication in multiple LAB hosts.
  • pPBT-GFP facilitates the use of pediocin and GFP as selectable markers.
  • The vector's stability and efficiency make it a valuable tool for genetic engineering of LAB for commercial applications.