Fig. 2: Screening, preparation, and characterization of MnLR. | Nature Communications

Fig. 2: Screening, preparation, and characterization of MnLR.

From: Manganese-engineered Lactobacillus Reuteri with enhanced antitumor and immunomodulatory activities for colorectal cancer prevention and treatment

Fig. 2: Screening, preparation, and characterization of MnLR.

a The schematic showing the Mn-enhanced proliferation and glycerol metabolism of L. reuteri. b, c The proliferation (measured by the optical density (OD) at 600 nm) and reuterin yield of L. reuteri cultured with different metal ions (Na+, Mg2+, K+, Ca2+, Mn2+, Fe2+, Cu2+, and Zn2+; 500 μM)) for 18 hours (for proliferation analysis) or 4 hours (for reuterin yield measurement), respectively (n = 4). d The proliferation and reuterin yield of L. reuteri cultured with Mn2+ at different concentrations. The inset, MRS platting assay for analyzing the L. reuteri amount after a 24-hour culture with Mn2+ (0, 300, and 800 μM) (n = 4). e The proliferation and reuterin yield of L. reuteri of 3rd or 50th generations cultured with or without Mn2+ (n = 5). f The proliferation and reuterin yield of L. reuteri cultured in SIF with or without Mn2+ (n = 5). g Principal component analysis (PCA) of gene profiles of untreated and Mn-treated L. reuteri. h The top nine enriched pathways in Mn-treated L. reuteri versus untreated control from Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. i The quantification of ribosomal proteins of L. reuteri with or without Mn2+ treatment by LC-MS/MS (n = 3). j The schematic presenting the fabrication of MnLR. k The representative scanning electronic microscopy (SEM) (Scale bar 1 μm), fluorescence (Scale bar 5 μm), and elemental mapping images of MnLR. CY 3 red fluorescence labels TA; and SYTO 9 green fluorescence labels nucleus. The proliferation and reuterin yield of raw L. reuteri and MnLR (l, n = 4; m, n = 3). n, o The representative images and corresponding colony counts of raw L. reuteri and MnLR cultured in SGF or SIF for 2 or 4 hours (n = 3). The “n” represents the number of biologically independent samples. Data are shown as mean ± SD; p-value (compared to Ctrl in b, c, i); one-way ANOVA (one-tailed), Tukey’s multiple comparisons test, or Student’s t-test for (e, i, l, m). Source data are provided as a Source Data file.

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