Construction and characterization of nisin-controlled expression vectors for use in Lactobacillus reuteri

Bioscience, Biotechnology, and Biochemistry
Chi-Ming WuTung-Ching Chung

Abstract

The Nisin-controlled gene expression (NICE) system, which was discovered in Lactococcus lactis, was adapted to Lactobacillus reuteri by ligating nisA promoter (PnisA) and nisRK DNA fragments into the Escherichia coli-Lb. reuteri shuttle vector pSTE32. This chimerical plasmid (pNICE) was capable of expressing the heterologous amylase gene (amyL) under nisin induction. Optimization of induction factors for this Lb. reuteri/pNICE system, including nisin concentration (viz. 50 ng/ml), growth phase of culture at which nisin be added (viz. at the early exponential phase), and the best time for analyzing the gene product after inoculation (viz. at the 3rd h), allowed the amylase product to be expressed in high amounts, constituting up to about 18% of the total intracellular protein. Furthermore, the signal peptide (SP) of amyL gene (SPamyL) from Bacillus licheniformis was ligated to the downstream of PnisA in pNICE, upgrading this vector to a NICE-secretion (NIES) level, which was then designated pNIES (Sec+, secretion positive). Characterization of pNIES using an amyL-SPDelta gene (amyL gene lacking its SP) as a reporter revealed the 3rd h after induction as the secretion peak of this system, at which the secretion efficiency and the...Continue Reading

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Citations

Nov 1, 2012·Microbial Cell Factories·Anna A ToymentsevaThorsten Mascher
Mar 8, 2011·Microbial Biotechnology·Tri DuongTodd R Klaenhammer
Apr 21, 2006·Journal of Microbiological Methods·Chi-Ming Wu, Tung-Ching Chung
Feb 27, 2016·Critical Reviews in Biotechnology·José Maria Landete
Jan 1, 2009·Microbial Biotechnology·Trygve BrautasetSvein Valla
Jan 19, 2019·Applied Microbiology and Biotechnology·Tina Vida Plavec, Aleš Berlec
Mar 5, 2020·Applied Microbiology and Biotechnology·Ángela Peirotén, José M Landete
Sep 25, 2017·Microbiology Spectrum·Laura Ortiz-Velez, Robert Britton

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