University of Oulu

Rasinkangas P, Tytgat HLP, Ritari J, Reunanen J, Salminen S, Palva A, Douillard FP and de Vos WM (2020) Characterization of Highly Mucus-Adherent Non-GMO Derivatives of Lacticaseibacillus rhamnosus GG. Front. Bioeng. Biotechnol. 8:1024. doi: 10.3389/fbioe.2020.01024

Characterization of highly mucus-adherent non-GMO derivatives of Lacticaseibacillus rhamnosus GG

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Author: Rasinkangas, Pia1; Tytgat, Hanne L. P.2; Ritari, Jarmo1;
Organizations: 1Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland
2Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands
3Functional Foods Forum, Faculty of Medicine, University of Turku, Turku, Finland
4Human Microbiome Research Program, Faculty of Medicine, University of Helsinki, Helsinki, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 6.1 MB)
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Language: English
Published: Frontiers Media, 2020
Publish Date: 2021-02-19


Lacticaseibacillus rhamnosus GG is one of the best studied lactic acid bacteria in the context of probiotic effects. L. rhamnosus GG 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 the spaCBA-srtC1 gene 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 of L. rhamnosus GG 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 the spaA and spaC genes 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 adherent L. rhamnosus GG 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 in L. rhamnosus GG and offer a variety of non-GMO derivatives of this key probiotic strain that are applicable in food products.

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Series: Frontiers in bioengineering and biotechnology
ISSN: 2296-4185
ISSN-E: 2296-4185
ISSN-L: 2296-4185
Volume: 8
Article number: 1024
DOI: 10.3389/fbioe.2020.01024
Type of Publication: A1 Journal article – refereed
Field of Science: 3111 Biomedicine
Funding: This project, including the efforts of PR, FD, JRe, and JRi, were funded by European Research Council (Grant 250172, Microbes Inside to WV) and Academy of Finland (Grants 252123 to FD, 299749 to JRe, and 137389 to WV). Work of HT and WV was partly funded by the SIAM Gravity Grant 024.002.002 and Spinoza Award of the Netherlands Organization for Scientific Research of WV.
Academy of Finland Grant Number: 299749
Detailed Information: 299749 (Academy of Finland Funding decision)
Copyright information: © 2020 Rasinkangas, Tytgat, Ritari, Reunanen, Salminen, Palva, Douillard and de Vos. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.