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Novel genetic modules encoding high-level antibiotic-free protein expression in probiotic lactobacilli

  • Lactobacilli are ubiquitous in nature, often beneficially associated with animals as commensals and probiotics, and are extensively used in food fermentation. Due to this close-knit association, there is considerable interest to engineer them for healthcare applications in both humans and animals, for which high-performance and versatile genetic parts are greatly desired. For the first time, we describe two genetic modules in Lactiplantibacillus plantarum that achieve high-level gene expression using plasmids that can be retained without antibiotics, bacteriocins or genomic manipulations. These include (i) a promoter, PtlpA, from a phylogenetically distant bacterium, Salmonella typhimurium, which drives up to 5-fold higher level of gene expression compared to previously reported promoters and (ii) multiple toxin-antitoxin systems as a self-contained and easy-to-implement plasmid retention strategy that facilitates the engineering of tuneable transient genetically modified organisms. These modules and the fundamental factors underlying their functionality that are described in this work will greatly contribute to expanding the genetic programmability of lactobacilli for healthcare applications.

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Document Type:Article
Author:Sourik DeyORCiD, Marc Blanch-Asensio, Sanjana Balaji Kuttae, Shrikrishnan SankaranORCiD
Parent Title (English):Microbial Biotechnology
First Page:1264
Last Page:1276
Year of first Publication:2023
Release Date:2023/06/22
Impact:05.830 (2021)
Funding Information:Deutsche Forschungsgemeinschaft. Grant Number 455063657; Collaborative Research Centre, SFB 1027 [Project # 200049484]; Leibniz Science Campus on Living Therapeutic Materials [LifeMat]
Groups:Bioprogrammierbare Materialien
DDC classes:500 Naturwissenschaften und Mathematik / 570 Biowissenschaften, Biologie
Open Access:Open Access
Signature:INM 2023/025
Licence (German):License LogoCreative Commons - CC BY-NC-ND - Namensnennung - Nicht kommerziell - Keine Bearbeitungen 4.0 International