Abstract
Simple SummaryConsiderable attention has been given to the species Limosilactobacillus reuteri regarding its probiotic potential. Here, the genome-scale metabolic model (GSMM) of L. reuteri KUB-AC5, namely iTN656 was developed and exploited to evaluate the metabolic capability of L. reuteri KUB-AC5 under various carbon sources. The simulated growth behaviors of iTN656 under single and double omission analysis promisingly identified 14 essential single nutrients and 2 essential double nutrients (pairwise glutamine-glutamate and asparagine-aspartate) for L. reuteri KUB-AC5 growth. Moreover, the integrated transcriptomic analysis using iTN656 allowed to probe the potential metabolic routes for enhancing growth of L. reuteri KUB-AC5 involved in sucrose uptake, nucleotide biosynthesis, urea cycle, and glutamine transporter. Overall, iTN656 provides a powerful systems biology platform for gaining insights into cell metabolism of lactic acid bacteria and guiding the rational development of synthetic media design for scale-up of probiotic production in industrial scale.Limosilactobacillus reuteri KUB-AC5 displays the hallmark features of probiotic properties for food and feed industries. Optimization of cultivation condition for the industrial production is important to reach cell concentration and cost reduction. Considering the strain-specific growth physiology, metabolic capability, and essential nutrients of L. reuteri KUB-AC5, the genome-scale metabolic model (GSMM) of L. reuteri KUB-AC5 was developed. Hereby, the GSMM of iTN656 was successfully constructed which contained 656 genes, 831 metabolites, and 953 metabolic reactions. The iTN656 model could show a metabolic capability under various carbon sources and guide potentially 14 essential single nutrients (e.g., vitamin B complex and amino acids) and 2 essential double nutrients (pairwise glutamine-glutamate and asparagine-aspartate) for L. reuteri KUB-AC5 growth through single and double omission analysis. Promisingly, the iTN656 model was further integrated with transcriptome data suggesting that putative metabolic routes as preferable paths e.g., sucrose uptake, nucleotide biosynthesis, urea cycle, and glutamine transporter for L. reuteri KUB-AC5 growth. The developed GSMM offers a powerful tool for multi-level omics analysis, enabling probiotic strain optimization for biomass overproduction on an industrial scale.
Highlights
Limosilactobacillus reuteri KUB-AC5, previously known as Lactobacillus reuteri KUBAC5 [1], belongs to lactic acid bacteria
L. reuteri is one of the predominant species serving for functional foods and feed supplements [2]
The metabolic network of L. reuteri JCM1112 [17] was selected as a template in this study since high genome similarity between L. reuteri KUB-AC5 and L. reuteri
Summary
Limosilactobacillus reuteri KUB-AC5, previously known as Lactobacillus reuteri KUBAC5 [1], belongs to lactic acid bacteria. The strain KUB-AC5 was originally isolated from chicken intestine displayed the hallmark features of probiotics [3]. This strain is able to produce antimicrobial substances which inhibited both of Gram-positive and Gramnegative bacteria, such as Escherichia coli, Salmonella Typhimurium, Salmonella Enteritidis, Bacillus cereus, Bacillus subtilis, Micrococcus luteus, and Staphylococcus aureus [3,4]. According to in vivo assay, strain KUB-AC5 could inhibit Salmonella infection in chicken and mice model [5,6]. This strain attenuated inflammation in both gut and spleen of mice model [6]. According to health benefit properties, L. reuteri KUB-AC5 is potentially useful for the development of new probiotic ingredient in the fields of food and feed stuffs at industrial scale
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