An Inducible Operon Is Involved in Inulin Utilization in Lactobacillus plantarum Strains, as Revealed by Comparative Proteogenomics and Metabolic Profiling.
The draft genomes of Lactobacillus plantarum strains isolated from Asian fermented foods, infant feces, and shrimp intestines were sequenced and compared to those of well-studied strains. Among 28 strains of L. plantarum, variations in the genomic features involved in ecological adaptation were elucidated. The genome sizes ranged from approximately 3.1 to 3.5 Mb, of which about 2,932 to 3,345 protein-coding sequences (CDS) were predicted. The food-derived isolates contained a higher number of carbohydrate metabolism-associated genes than those from infant feces. This observation correlated to their phenotypic carbohydrate metabolic profile, indicating their ability to metabolize the largest range of sugars. Surprisingly, two strains (P14 and P76) isolated from fermented fish utilized inulin. β-Fructosidase, the inulin-degrading enzyme, was detected in the supernatants and cell wall extracts of both strains. No activity was observed in the cytoplasmic fraction, indicating that this key enzyme was either membrane-bound or extracellularly secreted. From genomic mining analysis, a predicted inulin operon of fosRABCDXE, which encodes β-fructosidase and many fructose transporting proteins, was found within the genomes of strains P14 and P76. Moreover, pts1BCA genes, encoding sucrose-specific IIBCA components involved in sucrose transport, were also identified. The proteomic analysis revealed the mechanism and functional characteristic of the fosRABCDXE operon involved in the inulin utilization of L. plantarum The expression levels of the fos operon and pst genes were upregulated at mid-log phase. FosE and the LPXTG-motif cell wall anchored β-fructosidase were induced to a high abundance when inulin was present as a carbon source.



Inulin is a long-chain carbohydrate that may act as a prebiotic, which provides many health benefits to the host by selectively stimulating the growth and activity of beneficial bacteria in the colon. While certain lactobacilli can catabolize inulin, this has not yet been described for Lactobacillus plantarum, and an associated putative inulin operon has not been reported in this species. By using comparative and functional genomics, we showed that two L. plantarum strains utilized inulin and identified functional inulin operons in their genomes. The proteogenomic data revealed that inulin degradation and uptake routes, which related to the fosRABCDXE operon and pstBCA genes, were widely expressed among L. plantarum strains. The present work provides a novel understanding of gene regulation and mechanisms of inulin utilization in probiotic L. plantarum generating opportunities for synbiotic product development.
Department of Industrial Biotechnology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla, Thailand.|Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands.
Nirunya
N
Buntin
Department of Industrial Biotechnology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
Tipparat
T
Hongpattarakere
Department of Veterinary Biosciences, University of Helsinki, Helsinki, Finland.
Jarmo
J
Ritari
Department of Veterinary Biosciences, University of Helsinki, Helsinki, Finland.
François P
FP
Douillard
Institute of Biotechnology, University of Helsinki, Helsinki, Finland.
Lars
L
Paulin
Laboratory of Biochemistry, Wageningen University, Wageningen, The Netherlands.
Sjef
S
Boeren
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands.
Sudarshan A
SA
Shetty
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands willem.devos@wur.nl.|Department of Veterinary Biosciences, University of Helsinki, Helsinki, Finland.|Research Program Unit Immunobiology, Department of Bacteriology and Immunology, University of Helsinki, Helsinki, Finland.
Willem M
WM
de Vos
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007444:Inulin / Q000378:metabolism*; D048191:Lactiplantibacillus plantarum / Q000235:genetics* / Q000378:metabolism; D055442:Metabolome; D009876:Operon; D000071696:Proteogenomics
D001426:Bacterial Proteins; D007444:Inulin
fos operon; genomes; lactic acid bacteria; prebiotic; β-fructosidase