A Novel Class IIb Bacteriocin-Plantaricin EmF Effectively Inhibits Listeria monocytogenes and Extends the Shelf Life of Beef in Combination with Chitosan

Plantaricin EmF separated and identified from L. plantarum 163 was a novel class IIb bacteriocin. The molecular masses of plantaricin Em and F were 1638 and 3702 Da, respectively, with amino acid sequences FNRGGYNFGKSVRH and VFHAYSARGVRNNYKSAVGPADWVISAVRGFIHG, respectively. Plantaricin EmF not only exhibited broad-pH adaptability and thermostability but also showed high efficiency and broad-spectrum antibacterial activity. Its mode of action on L. monocytogenes damaged cell membrane integrity, resulting in the leakage of cytoplasm, changes in cell structure and morphology, and ultimately cell death. Additionally, plantaricin EmF inactivated L. monocytogenes in beef, effectively improving the quality indices of beef, thereby extending its shelf life, especially in combination with chitosan. Plantaricin EmF + 1.0% chitosan extended the shelf life of beef to 15 d, demonstrating its potential application value to replace chemical preservatives to control food-borne pathogenic microorganisms and extend the shelf life of meat and meat products in agriculture and the food industry

Lactobacillus acidophilus NX2‑6 Improved High-Fat Diet-Induced Glucose Metabolism Disorder Independent of Promotion of Insulin Secretion in Mice

High-fat diet (HFD) contributes to metabolic inflammation and glucose metabolism disorder, thereby resulting in the pathogenesis of metabolic syndrome. Accumulating evidence has revealed that some probiotics could improve HFD-induced metabolic inflammation and glucose metabolism disorder. Our previous study has discovered that Lactobacillus acidophilus NX2-6 exhibited in vitro lipid-lowering, antioxidative, and anti-inflammatory activities. This study mainly investigated whether L. acidophilus NX2-6 improved HFD-induced glucose metabolism disorder. The results exhibited that L. acidophilus NX2-6 effectively reduced blood glucose levels and improved glucose tolerance by activating the insulin signaling pathway, promoting glucose uptake, glycolysis, and intestinal gluconeogenesis and suppressing hepatic gluconeogenesis, independent of regulation of glycogen synthesis in the liver and muscle. Enhanced insulin sensitivity was associated with L. acidophilus NX2-6-mediated suppression of inflammatory cascades in the target organs. Meanwhile, L. acidophilus NX2-6 also improved hepatic energy metabolism via the FGF21/AMPKα/PGC-1α/NRF1 pathway. However, L. acidophilus NX2-6 did not affect apoptosis, pyroptosis, inflammation, and endoplasmic reticulum stress in the pancreas of HFD-fed mice. In conclusion, our results indicated that L. acidophilus NX2-6 improved glucose metabolism disorder through enhancing insulin sensitivity, suppressing metabolic inflammation, and promoting energy expenditure

TetR-Type Regulator Lp_2642 Positively Regulates Plantaricin EF Production Based on Genome-Wide Transcriptome Sequencing of Lactiplantibacillus plantarum 163

Whole-genome and transcriptome sequences of Lactiplantibacillus plantarum 163 are provided. There was one circular chromosome and four circular plasmids, with sizes of 3,131,367; 56,674; 49,140; 43,628; and 36,387 bp, respectively, in L. plantarum 163. The regulator Lp_2642 was selected from the genome data, the overexpression of which increased the transcriptional levels of related genes in plantaricin EF biosynthesis and enhanced plantaricin EF production. Its production was 17.30 mg/L in 163 (Lp_2642), which was 1.29-fold higher than that of the original strain. The regulation mechanism demonstrated that Lp_2642 can bind to three sites of plnA promoter, which enhances its transcription and expression, thereby increasing plantaricin EF production. Amino acids Asn-100, Asn-64, and Thr-69 may play a key role in the binding of Lp_2642. These results provide a novel strategy for mass production of plantaricin EF, which facilitates its large-scale production and application in the agriculture and food industries as a preservative

Additional file 8 of Transcriptomic analysis reveals that Bacillomycin D-C16 induces multiple pathways of disease resistance in cherry tomato

Additional file 8: Table S8. Table of transcription factors after Bacillomycin D-C16 treatment versus control at time 12 h

Table1_Improved catalytic performance and molecular insight for lipoxygenase from Enterovibrio norvegicus via directed evolution.DOCX

Lipoxygenase (LOX) holds significant promise for food and pharmaceutical industries. However, albeit its application has been hampered by low catalytic activity and suboptimal thermostability. To address the drawbacks, a directed evolution strategy was explored to enhance the catalytic activity and thermostability of LOX from Enterovibrio norvegicus (EnLOX) for the first time. After two rounds of error-prone polymerase chain reaction (error-prone PCR) and one generations of sequential DNA shuffling, all of four different mutants showed a significant increase in the specific activity of EnLOX, ranging from 132.07 ± 9.34 to 330.17 ± 18.54 U/mg. Among these mutants, D95E/T99A/A121H/S142N/N444W/S613G (EAHNWG) exhibited the highest specific activity, which was 8.25-fold higher than the wild-type enzyme (WT). Meanwhile, the catalytic efficiency (Kcat/Km) of EAHNWG was also improved, which was 13.61 ± 1.67 s−1 μM−1, in comparison to that of WT (4.83 ± 0.38 s−1 μM−1). In addition, mutant EAHNWG had a satisfied thermostability with the t1/2,50 °C value of 6.44 ± 0.24 h, which was 0.4 h longer than that of the WT. Furthermore, the molecular dynamics simulation and structural analysis demonstrated that the reduction of hydrogen bonds number, the enhancement of hydrophobic interactions in the catalytic pocket, and the improvement of flexibility of the lid domain facilitated structural stability and the strength of substrate binding capacity for improved thermal stability and catalytic efficiency of mutant LOX after directed evolution. Overall, these results could provide the guidance for further enzymatic modification of LOX with high catalytic performance for industrial application.</p

Purification, Characterization, and Mode of Action of Plantaricin GZ1-27, a Novel Bacteriocin against <i>Bacillus cereus</i>

Bacillus cereus is an opportunistic pathogen that causes foodborne diseases. We isolated a novel bacteriocin, designated plantaricin GZ1-27, and elucidated its mode of action against B. cereus. Plantaricin GZ1-27 was purified using ammonium sulfate precipitation, gel-filtration chromatography, and RP-HPLC. MALDI-TOF/MS revealed that its molecular mass was 975 Da, and Q-TOF-MS/MS analysis predicted the amino acid sequence as VSGPAGPPGTH. Plantaricin GZ1-27 showed thermostability and pH stability. The antibacterial mechanism was investigated using flow cytometry, confocal laser-scanning microscopy, scanning and transmission electron microscopy, and RT-PCR, which revealed that GZ1-27 increased cell membrane permeability, triggered K+ leakage and pore formation, damaged cell membrane integrity, altered cell morphology and intracellular organization, and reduced the expression of genes related to cytotoxin production, peptidoglycan synthesis, and cell division. These results suggest that plantaricin GZ1-27 effectively inhibits B. cereus at both the cellular and the molecular levels and is a potential natural food preservative targeting B. cereus

Knockout of <i>rapC</i> Improves the Bacillomycin D Yield Based on <i>De Novo</i> Genome Sequencing of Bacillus amyloliquefaciens fmbJ

Bacillus amyloliquefaciens, a Gram-positive and soil-dwelling bacterium, could produce secondary metabolites that suppress plant pathogens. In this study, we provided the whole genome sequence results of B. amyloliquefaciens fmbJ, which had one circular chromosome of 4 193 344 bp with 4249 genes, 87 tRNA genes, and 27 rRNA genes. In addition, fmbJ was found to contain several gene clusters of antimicrobial lipopeptides (bacillomycin D, surfactin, and fengycin), and bacillomycin D homologues were further comprehensively identified. To clarify the influence of <i>rapC</i> regulating the synthesis of lipopeptide on the yield of bacillomycin D, <i>rapC</i> gene in fmbJ was successfully deleted by the marker-free method. Finally, it was found that the deletion of <i>rapC</i> gene in fmbJ significantly improved bacillomycin D production from 240.7 ± 18.9 to 360.8 ± 30.7 mg/L, attributed to the increased the expression of bacillomycin D synthesis-related genes through enhancing the transcriptional level of <i>comA</i>, <i>comP</i>, and <i>phrC</i>. These results showed that the production of bacillomycin D in B. amyloliquefaciens fmbJ might be regulated by the RapC–PhrC system. The findings are expected to advance further agricultural application of Bacillus spp. as a promising source of natural bioactive compounds

Additional file 4 of Transcriptomic analysis reveals that Bacillomycin D-C16 induces multiple pathways of disease resistance in cherry tomato

Additional file 4: Table S4. Table of GO enrichment analysis after Bacillomycin D-C16 treatment versus control at time 12 h

Additional file 1 of Transcriptomic analysis reveals that Bacillomycin D-C16 induces multiple pathways of disease resistance in cherry tomato

Additional file 1: Table S1. Overview of the transcriptome sequencing dataset and quality check

Additional file 3 of Transcriptomic analysis reveals that Bacillomycin D-C16 induces multiple pathways of disease resistance in cherry tomato

Additional file 3: Table S3. Table of RNA-seq data after Bacillomycin D-C16 treatment versus control at time 24 h