Abstract
The manufacturing processes of commercial probiotic strains may be affected in different ways in the attempt to optimize yield, costs, functionality, or stability, influencing gene expression, protein patterns, or metabolic output. Aim of this work is to compare different samples of a high concentration (450 billion bacteria) multispecies (8 strains) formulation produced at two different manufacturing sites, United States of America (US) and Italy (IT), by applying a combination of functional proteomics, metabolomics, and in vivo analyses. Several protein-profile differences were detected between IT- and US-made products, with Lactobacillus paracasei, Streptococcus thermophilus, and Bifidobacteria being the main affected probiotics/microorganisms. Performing proton nuclear magnetic spectroscopy (1H-NMR), some discrepancies in amino acid, lactate, betaine and sucrose concentrations were also reported between the two products. Finally, we investigated the health-promoting and antiaging effects of both products in the model organism Caenorhabditis elegans. The integration of omics platforms with in vivo analysis has emerged as a powerful tool to assess manufacturing procedures.
Highlights
Probiotics are defined by the Food and Agriculture Organization of the United Nation/World Health Organization (FAO/WHO) as “live microorganisms, which when administered in adequate amounts, confer a health benefit on the host” [1]
A total of 3685 proteins at a false discovery rate of 1% were identified by nano LC-MS/MS analysis; among these, 2553 proteins could be consistently quantified in all the replicates of at least one sample (Table S2)
Quantitative differences occurring among proteomic profiles of different lots were detected by comparing their protein intensities with those from the United States of America (US)-4 product that was used as control group, being the most recent lot produced at Danisco/Dupont
Summary
Probiotics are defined by the Food and Agriculture Organization of the United Nation/World Health Organization (FAO/WHO) as “live microorganisms, which when administered in adequate amounts, confer a health benefit on the host” [1]. A number of variables are involved in the industrial production of bacterial species, i.e., growth conditions, substrates, cryoprotectants and storage conditions, all of which affect strains’ probiotic properties by influencing gene expression, protein patterns, and/or metabolic output. Genomics is a powerful tool to identify bacterial genus/species and to detect the presence of drug resistance and virulence genes, but it cannot ascertain whether or not genetically related strains have the same probiotic properties. Changes in the industrial processes have been shown to affect protein composition, chemical characteristics of the cell wall, and anti-pathogen activity of the probiotic strain Lactobacillus rhamnosus Lcr35 [2]. Grzeskowiak studied several L. rhamnosus GG isolates from different probiotic products and confirmed that some beneficial properties were lost depending on the manufacturing site, notwithstanding the genetic identity of the strains tested [3]. Some publications discussed the equivalence of the VSL#3 probiotic formulation produced in two different manufacturing facilities [4,5,6]
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