Construction of a Shuttle Expression Vector for Lactic Acid Bacteria

Overview

Journal J Genet Eng Biotechnol

Specialty Biotechnology

Date 2019 Nov 19

PMID 31736018

Citations 4

Authors

Tejinder Kaur

Praveen P Balgir

Baljinder Kaur

Affiliations

Soon will be listed here.

Abstract

Background: Lactic acid bacteria (LAB) are a diverse group of Gram-positive bacteria, which are widely distributed in various diverse natural habitats. These are used in a variety of industrial food fermentations and carry numerous traits with utmost relevance to the food industry. Genetic engineering has emerged as an effective means to improve and enhance the potential of commercially important bacterial strains. However, the biosafety of recombinant systems is an important concern during the implementation of such technologies on an industrial scale. In order to overcome this issue, cloning and expression systems have been developed preferably from fully characterized and annotated LAB plasmids encoding genes with known functions.

Results: The developed shuttle vector pPBT-GFP contains two theta-type replicons with a copy number of 4.4 and 2.8 in Pediococcus acidilactici MTCC 5101 and Lactobacillus brevis MTCC 1750, respectively. Antimicrobial "pediocin" produced by P. acidilactici MTCC 5101 and green fluorescent protein (GFP) of Aequorea victoria were successfully expressed as selectable markers. Heterologous bile salt hydrolase (BSH) from Lactobacillus fermentum NCDO 394 has been efficiently expressed in the host strains showing high specific activity of 126.12 ± 10.62 in P. acidilactici MTCC 5101 and 95.43 ± 4.26 in the case of L. brevis MTCC 1750, towards glycine-conjugated bile salts preferably as compared to taurine-conjugated salts.

Conclusion: The present article details the development of a LAB/LAB shuttle expression vector pPBT-GFP, capable of replication in LAB hosts, P. acidilactici MTCC 5101, and L. brevis MTCC 1750. Pediocin and GFP have been used as selectable markers with the efficient production of heterologous extracellular bile salt hydrolase. Thus, the constructed vector pPBT-GFP, with its ability to replicate in multiple hosts, low copy number, and stability in host cells, may serve as an ideal tool for improving LAB strains of commercial value using genetic engineering.

Citing Articles

Plasmid-Based Gene Expression Systems for Lactic Acid Bacteria: A Review.

Kazi T, Acharya A, Mukhopadhyay B, Mandal S, Arukha A, Nayak S Microorganisms. 2022; 10(6).

PMID: 35744650 PMC: 9229153. DOI: 10.3390/microorganisms10061132.

Genome-wide high-throughput signal peptide screening via plasmid pUC256E improves protease secretion in Lactiplantibacillus plantarum and Pediococcus acidilactici.

Chen B, Loo B, Cheng Y, Song P, Fan H, Latypov O BMC Genomics. 2022; 23(1):48.

PMID: 35021997 PMC: 8756648. DOI: 10.1186/s12864-022-08292-3.

Production of Functional Buttermilk and Soymilk Using BD16 ().

Sharma A, Noda M, Sugiyama M, Ahmad A, Kaur B Molecules. 2021; 26(15).

PMID: 34361824 PMC: 8347781. DOI: 10.3390/molecules26154671.

Correction to: Construction of a shuttle expression vector for lactic acid bacteria.

Kaur T, Balgir P, Kaur B J Genet Eng Biotechnol. 2020; 18(1):38.

PMID: 32749538 PMC: 7403355. DOI: 10.1186/s43141-020-00056-4.

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