Characterization of Highly Mucus-Adherent Non-GMO Derivatives of Lacticaseibacillus rhamnosus GG

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Rasinkangas , P , Tytgat , H L P , Ritari , J , Reunanen , J , Salminen , S , Palva , A , Douillard , F P & de Vos , W M 2020 , ' Characterization of Highly Mucus-Adherent Non-GMO Derivatives of Lacticaseibacillus rhamnosus GG ' , Frontiers in Bioengineering and Biotechnology , vol. 8 , 1024 . https://doi.org/10.3389/fbioe.2020.01024

Title: Characterization of Highly Mucus-Adherent Non-GMO Derivatives of Lacticaseibacillus rhamnosus GG
Author: Rasinkangas, Pia; Tytgat, Hanne L. P.; Ritari, Jarmo; Reunanen, Justus; Salminen, Seppo; Palva, Airi; Douillard, Francois P.; de Vos, Willem M.
Other contributor: University of Helsinki, Veterinary Biosciences
University of Helsinki, Veterinary Biosciences
University of Helsinki, Department of Food and Nutrition
University of Helsinki, Departments of Faculty of Veterinary Medicine
University of Helsinki, Food Hygiene and Environmental Health
University of Helsinki, Department of Bacteriology and Immunology











Date: 2020-08-19
Language: eng
Number of pages: 11
Belongs to series: Frontiers in Bioengineering and Biotechnology
ISSN: 2296-4185
DOI: https://doi.org/10.3389/fbioe.2020.01024
URI: http://hdl.handle.net/10138/319679
Abstract: Lacticaseibacillus rhamnosusGG is one of the best studied lactic acid bacteria in the context of probiotic effects.L. rhamnosusGG has been shown to prevent diarrhea in children and adults and has been implicated to have mitigating or preventive effects in several disorders connected to microbiota dysbiosis. The probiotic effects are largely attributed to its adhesive heterotrimeric sortase-dependent pili, encoded by thespaCBA-srtC1gene cluster. Indeed, the strain-specific SpaCBA pili have been shown to contribute to adherence, biofilm formation and host signaling. In this work we set out to generate non-GMO derivatives ofL. rhamnosusGG that adhere stronger to mucus compared to the wild-type strain using chemical mutagenesis. We selected 13 derivatives that showed an increased mucus-adherent phenotype. Deep shotgun resequencing of the strains enabled division of the strains into three classes, two of which revealed SNPs (single nucleotide polymorphisms) in thespaAandspaCgenes encoding the shaft and tip adhesive pilins, respectively. Strikingly, the other class derivatives demonstrated less clear genotype - phenotype relationships, illustrating that pili biogenesis and structure is also affected by other processes. Further characterization of the different classes of derivatives was performed by PacBio SMRT sequencing and RNAseq analysis, which resulted in the identification of molecular candidates driving pilin biosynthesis and functionality. In conclusion, we report on the generation and characterization of three classes of strongly adherentL. rhamnosusGG derivatives that show an increase in adhesion to mucus. These are of special interest as they provide a window on processes and genes driving piliation and its control inL. rhamnosusGG and offer a variety of non-GMO derivatives of this key probiotic strain that are applicable in food products.
Subject: pili
mucus
Lacticaseibacillus rhamnosusGG
adhesion
probiotics
non-GMO
ANTIBIOTIC-ASSOCIATED DIARRHEA
SPAC PILIN SUBUNIT
FUNCTIONAL-ANALYSIS
DOUBLE-BLIND
GG
ADHESION
PILUS
CHILDREN
INFECTIONS
VIRULENCE
11832 Microbiology and virology
217 Medical engineering
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