Genome Sequence of Lactobacillus mucosae LM1, Isolated from Piglet Feces

Abstract

e Lactobacillus mucosae LM1, isolated from stool samples of a healthy piglet, displays good in vitro mucin adhesion and antimicrobial activity against pathogenic bacteria. To elucidate its antimicrobial effects and to find its epithelial cell and mucin adhesion genes, the genomic sequence of L. mucosae LM1 was investigated.L actobacillus mucosae, found in the mammalian gastrointestinal tract, has been shown to have the ability to adhere to mucosal surfaces (2,3,9). Previous reports have identified L. mucosae as having the ability to attach tightly to the epithelium of the human intestine and to produce antimicrobials and a biofilm when exposed to the physiological conditions of the gut (4). The adhesion of lactobacilli to gastrointestinal mucus makes them a good choice for probiotics since it increases their ability to colonize the gut efficiently, modulate the intestinal immune system, and inhibit pathogenic bacteria (1, 4, 9).L. mucosae LM1 was isolated from stool samples from a healthy piglet (6). Preliminary trials regarding the adhesion and antibacterial activity of L. mucosae LM1 demonstrated good mucin-binding activity in vitro and antibacterial activity against pathogenic bacteria. The genome of L. mucosae LM1 was determined using a Roche 454 GS FLX sequencer and Illumina GA IIx platform. All reads were assembled into 55 contigs by de novo assembly. The initial draft assembly was prepared from the libraries of 22,092,187 reads (950ϫ coverage) using Newbler Assembler 2.3 (Roche), CLC Genomics Workbench 4.8 (CLCbio), and CodonCode Aligner (CodonCode Co.). A functional annotation was performed by the Rapid Annotation using Subsystem Technology (RAST) server and BLASTP-based comparisons with the KEGG and COG databases.The draft genome of L. mucosae LM1 included 2,213,697 bp with a 45.87% GϩC content, 2,039 protein-coding genes, and 56 tRNA-encoding genes. Functions were assigned to 64.6% (1,318) of the total coding sequences; 8.7% (428) were found to be hypothetical proteins that are unique to this strain. A phylogenetic tree produced from the 16S rRNA genes revealed that strain LM1 is most closely related to L. mucosae CCUG 43169 (8). Likewise, 16S rRNA analysis showed strong homology to other Lactobacillus species with completed genomes, including Lactobacillus reuteri DSM 20016 (NCBI reference NC_009513.1), with 94% similarity, and Lactobacillus fermentum IF03956 (GenBank reference AP008937.1), with 95% similarity. The 16S rRNA gene sequence was extracted from whole-genome shotgun assemblies derived from the EzTaxon-e database (5).An analysis of the L. mucosae LM1 genome revealed that LM1 has a specific mucus-binding protein (mub) gene (LBLM1_04370), which showed 95% coverage and 93% similarity to the best-matched L. reuteri mub gene. The mucus-binding activity induced by this mub gene has antimicrobial effects through cell surface protection (7,8). Moreover, the L. mucosae LM1 genome includes a putative ABC transporter and adhesin-like protein (LBLM1_10110) with significant homology (100% coverage and 93...

In vitroevaluation of the mucin-adhesion ability and probiotic potential ofLactobacillus mucosaeLM1

“…Preliminary genome analysis of our strain, Lact. mucosae LM1, supports this finding as we have identified numerous adhesion-related genes (Lee et al 2012) in its genome assembly. Likewise, other probiotic genes were also identified, such as bile salt hydrolases and beta-galactosidases, which lower cholesterol levels ) and ameliorate lactose intolerance in the host (De Vrese et al 2001), respectively, noting its probiotic potential.…”

“…; Lee et al . ). These proteins may bind specifically and adhere to mucus because of O‐glycan chains that contain Lewis‐type or ABO blood group antigens (Van Tassell and Miller ).…”

“…The primary Lactobacillus strain used in this study was Lact. mucosae LM1 (GenBank accession number, AHIT00000000), which was isolated from piglet faeces (Lee et al 2012). Other lactobacilli used in this study for probiotic adhesion assessment and cell-surface characterization included Lact.…”

“…mucosae LM1, supports this finding as we have identified numerous adhesion‐related genes (Lee et al . ) in its genome assembly. Likewise, other probiotic genes were also identified, such as bile salt hydrolases and beta‐galactosidases, which lower cholesterol levels (Chae et al .…”

Increased propionate concentration inLactobacillus mucosae-fermented wet brewers grains and duringin vitrorumen fermentation

“…Lactobacillus mucosae is a species of lactic acid bacteria (LAB) that exhibits antimicrobial activity against pathogenic bacteria (Lee et al . ), as well as the ability to improve intestinal barrier function by decreasing intestinal epithelial permeability (Smokvina and Degivry ).…”