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

BACKGROUND: Bacillomycin D-C16 can induce resistance in cherry tomato against pathogens; however, the underlying molecular mechanism is poorly understood. Here, the effect of Bacillomycin D-C16 on induction of disease resistance in cherry tomato was investigated using a transcriptomic analysis. RESULTS: Transcriptomic analysis revealed a series of obvious enrichment pathways. Bacillomycin D-C16 induced phenylpropanoid biosynthesis pathways and activated the synthesis of defense-related metabolites including phenolic acids and lignin. Moreover, Bacillomycin D-C16 triggered a defense response through both hormone signal transduction and plant-pathogen interactions pathways, and increased the transcription of several transcription factors (e.g., AP2/ERF, WRKY and MYB). These transcription factors might contribute to the further activated the expression of defense-related genes (PR1, PR10 and CHI) and stimulated the accumulation of H2O2. CONCLUSION: Bacillomycin D-C16 can induce resistance in cherry tomato by activating the phenylpropanoid biosynthesis pathway, hormone signal transduction pathway and plant-pathogen interactions pathway, thus activating comprehensive defense reaction against pathogen invasion. These results provided a new insight into the bio-preservation of cherry tomato by the Bacillomycin D-C16.

In-Field Tobacco Leaf Maturity Detection with an Enhanced MobileNetV1: Incorporating a Feature Pyramid Network and Attention Mechanism.

The maturity of tobacco leaves plays a decisive role in tobacco production, affecting the quality of the leaves and production control. Traditional recognition of tobacco leaf maturity primarily relies on manual observation and judgment, which is not only inefficient but also susceptible to subjective interference. Particularly in complex field environments, there is limited research on in situ field maturity recognition of tobacco leaves, making maturity recognition a significant challenge. In response to this problem, this study proposed a MobileNetV1 model combined with a Feature Pyramid Network (FPN) and attention mechanism for in situ field maturity recognition of tobacco leaves. By introducing the FPN structure, the model fully exploits multi-scale features and, in combination with Spatial Attention and SE attention mechanisms, further enhances the expression ability of feature map channel features. The experimental results show that this model, with a size of 13.7 M and FPS of 128.12, performed outstandingly well on the task of field maturity recognition of tobacco leaves, achieving an accuracy of 96.3%, superior to classical models such as VGG16, VGG19, ResNet50, and EfficientNetB0, while maintaining excellent computational efficiency and small memory footprint. Experiments were conducted involving noise perturbations, changes in environmental brightness, and occlusions to validate the model's robustness in dealing with the complex environments that may be encountered in actual applications. Finally, the Score-CAM algorithm was used for result visualization. Heatmaps showed that the vein and color variations of the leaves provide key feature information for maturity recognition. This indirectly validates the importance of leaf texture and color features in maturity recognition and, to some extent, enhances the credibility of the model. The model proposed in this study maintains high performance while having low storage requirements and computational complexity, making it significant for in situ field maturity recognition of tobacco leaves.

Acetate and auto-inducing peptide are independent triggers of quorum sensing in Lactobacillus plantarum.

The synthesis of plantaricin in Lactobacillus plantarum is regulated by quorum sensing. However, the nature of the extra-cytoplasmic (EC) sensing domain of the histidine kinase (PlnB1) and the ability to recognize the auto-inducing peptide PlnA1 is not known. We demonstrate the key motif Ile-Ser-Met-Leu of auto-inducing peptide PlnA1 binds to the hydrophobic region Phe-Ala-Ser-Gln-Phe of EC loop 2 of PlnB1 via hydrophobic interactions and hydrogen bonding. Moreover, we identify a new inducer, acetate, that regulates the synthesis of plantaricin by binding to a positively charged region (Arg-Arg-Tyr-Ser-His-Lys) in loop 4 of PlnB1 via electrostatic interaction. The side chain of Phe143 on loop 4 determined the specificity and affinity of PlnB1 to recognize acetate. PlnA1 activates quorum sensing in log phase growth and acetate in stationary phase to maintain the synthesis of plantaricin under conditions of reduced growth. Acetate activation of PlnB was also evident in four types of PlnB present in different Lb. plantarum strains. Finally, we proposed a model to explain the developmental regulation of plantaricin synthesis by PlnA and acetate. These results have potential applications in improving food fermentation and bacteriocin production.

Plantaricin A reverses resistance to ciprofloxacin of multidrug-resistant Staphylococcus aureus by inhibiting efflux pumps.

Overexpression of Staphylococcus aureus efflux pumps is commonly associated with antibiotic resistance, causing conventional antibiotics to be unsuccessful in combating multidrug-resistant bacterial infections. Reducing the activity of the efflux pump is an urgently required to tackle this problem. Here, we found that plantaricin A (PlnA), an antimicrobial peptide derived from Lactobacillus plantarum, had a synergistic effect with ciprofloxacin (CIP), reducing the IC90 of CIP by eight times. Subsequently, changes in membrane permeability, membrane potential, and reactive oxygen species (ROS) were determined; changes that did not explain the synergistic effect were previously observed. Ethidium bromide intake and efflux experiments showed that PlnA inhibited the function of the efflux pump by binding it and altering the structure of MepA, NorA, and LmrS. Then, a series of PlnA mutants were designed to explore the underlying mechanism; they showed that the charge and foaming of PlnA were the predominant factors affecting the structure of NorA. In a skin wound infection model, PlnA significantly reduced the dose of CIP, relieved inflammation, and promoted wound healing, indicating that PlnA and CIP synergy persisted in vivo. Overall, PlnA reduced the use of CIP for combination therapy, and allowing the continued used of CIP to kill MDR S. aureus. Multidrug-resistant Staphylococcus aureus threatens our life as a tenacious pathogen, which causes infections in hospitals, communities and animal husbandry. Various studies have showed that efflux pump inhibitors (EPIs) have been considered potential therapeutic agents for rejuvenating the activity of antibiotics. Unfortunately, small molecule EPIs exhibit several side effects that limit their use for clinical application. The present study showed a new EPI (plantaricin A) produced by Lactobacillus plantarum, which has low cytotoxicity and haemolysis and powerful inhibitory activity on efflux pumps. Therefore, it helps the design of new EPIs and controls the infection of MDR S. aureus.

Plantaricin A, Derived from Lactiplantibacillus plantarum, Reduces the Intrinsic Resistance of Gram-Negative Bacteria to Hydrophobic Antibiotics.

The outer membrane of Gram-negative bacteria is one of the major factors contributing to the development of antibiotic resistance, resulting in a lack of effectiveness of several hydrophobic antibiotics. Plantaricin A (PlnA) intensifies the potency of antibiotics by increasing the permeability of the bacterial outer membrane. Moreover, it has been proven to bind to the lipopolysaccharide of Escherichia coli via electrostatic and hydrophobic interactions and to interfere with the integrity of the bacterial outer membrane. Based on this mechanism, we designed a series of PlnA1 analogs by changing the structure, hydrophobicity, and charge to enhance their membrane-permeabilizing ability. Subsequent analyses revealed that among the PlnA1 analogs, OP4 demonstrated the highest penetrating ability, weaker cytotoxicity, and a higher therapeutic index. In addition, it decelerated the development of antibiotic resistance when the E. coli cells were continuously exposed to sublethal concentrations of erythromycin and ciprofloxacin for 30 generations. Further in vivo studies in mice with sepsis showed that OP4 heightens the potency of erythromycin against E. coli and relieves inflammation. In summary, our results showed that the PlnA1 analogs investigated in the present study, especially OP4, reduce the intrinsic antibiotic resistance of Gram-negative pathogens and expand the antibiotic sensitivity spectrum of hydrophobic antibiotics in Gram-negative bacteria. IMPORTANCE Antibiotic resistance is a global health concern due to indiscriminate use of antibiotics, resistance transfer, and intrinsic resistance of certain Gram-negative bacteria. The asymmetric bacterial outer membrane prevents the entry of hydrophobic antibiotics and renders them ineffective. Consequently, these antibiotics could be employed to treat infections caused by Gram-negative bacteria, after increasing their outer membrane permeability. As PlnA reportedly penetrates outer membranes, we designed a series of PlnA1 analogs and proved that OP4, one of these antimicrobial peptides, effectively augmented the permeability of the bacterial outer membrane. Furthermore, OP4 effectively improved the potency of erythromycin and alleviated inflammatory responses caused by Escherichia coli infection. Likewise, OP4 curtailed antibiotic resistance development in E. coli, thereby prolonging exposure to sublethal antibiotic concentrations. Thus, the combined use of hydrophobic antibiotics and OP4 could be used to treat infections caused by Gram-negative bacteria by decreasing their intrinsic antibiotic resistance.

Lacticaseibacillus rhamnosus Fmb14 prevents purine induced hyperuricemia and alleviate renal fibrosis through gut-kidney axis.

Hyperuricemia is a critical threat to human health, and conventional medical treatment only aims to treat acute gouty arthritis. Purine diet-mediated chronic hyperuricemia and related syndromes are neglected in clinical therapeutics. In this study, the prevention ability of Lacticaseibacillus rhamnosus Fmb14, screened from Chinese yogurt, was evaluated in chronic purine-induced hyperuricemia (CPH) mice. After 12 weeks of Fmb14 administration, serum uric acid (SUA) in CPH mice decreased by 36.8 %, from 179.1 to 113.2 µmol/L, and the mortality rate decreased from 30 % to 10 %. The prevention role of Fmb14 in CPH was further investigated, and the reduction of uric acid by Fmb14 was attributed to the reduction of XOD (xanthine oxidase) in the liver and URAT1 in the kidney, as well the promotion of ABCG2 in the colon. Fmb14 administration Increased ZO-1 and Occludin expression in the colon and decreased fibrosis degree in the kidney indicated that Fmb14 administration had preventive effects through the gut-kidney axis in CPH. In specific, Fmb14 administration upregulated the diversity of gut microbiota, increased short-chain fatty acids (SCFA) by 35 % in colon materials and alleviated the inflammatory response by reducing biomarkers levels of IL-1ß, IL-18 and TNF-α at 11.6 %, 21.7 % and 26.5 % in serum, compared to CPH group, respectively. Additionally, 16 S rRNA sequencing showed 31.5 % upregulation of Prevotella, 20.5 % and 21.6 % downregulation of Ruminococcus and Suterella at the genus level, which may be a new gut microbial marker in hyperuricemia. In conclusion, Fmb14 ameliorated CPH through the gut-kidney axis, suggesting a new strategy to prevent hyperuricemia.

An endolysin Salmcide-p1 from bacteriophage fmb-p1 against gram-negative bacteria.

AIMS: A novel endolysin Salmcide-p1 was developed as a promising candidate of new preservative and a supplement to effective enzyme preparations against gram-negative bacterial contaminations. METHODS AND RESULTS: Salmcide-p1 was identified by complementing the genomic sequence of a virulent Salmonella phage fmb-p1. Salmcide-p1 of 112 µg ml-1 could quickly kill Salmonella incubated with 100 mmol l-1 EDTA, with no haemolytic activity. Meanwhile, Salmcide-p1 had a high activity of lysing Salmonella cell wall peptidoglycan. At different temperatures (4-75°C), pH (4-11) and NaCl concentration (10-200 mmol l-1 ), the relative activity of Salmcide-p1 was above 60%. At 4°C, the combination of Salmcide-p1 and EDTA-2Na could inhibit the number of Salmonella Typhimurium CMCC 50115 in skim milk to less than 4 log CFU ml-1 by 3 days, and the number of Shigella flexneri CMCC 51571 was lower than 4 log CFU ml-1 by 9 days. CONCLUSIONS: Salmcide-p1 had a wide bactericidal activity against gram-negative bacteria and showed a broader anti-Salmonella spectrum than the phage fmb-p1. The combination strategy of Salmcide-p1 and EDTA-2Na could significantly inhibit the growth of gram-negative bacteria inoculated in skim milk. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteriophage endolysin as an antibacterial agent is considered to be a new strategy against bacterial contamination.

Surfactin-oleogel with therapeutic potential for inflammatory acne vulgaris induced by Propionibacterium acnes.

Accumulating evidence suggested that suppression of Propionibacterium acnes-induced inflammation was a promising strategy to alleviate acne vulgaris. This study evaluated the alleviating effect of surfactin-oleogel on P. acnes-induced inflammatory acne vulgaris in mice. Epidermis morphology and histopathological examination showed that surfactin-oleogel effectively ameliorated the P. acnes-induced epidermis swelling and erythema. Surfactin-oleogel reduced the epidermis thickness to 48.52% compared to the model control group. The colony of P. acnes in the epidermis was decreased by 1 log CFU/mL after receiving surfactin-oleogel treatment. Furthermore, surfactin-oleogel attenuated oxidative stress in the epidermis by increasing the activities of superoxide dismutase, catalase, and glutathione peroxidase. In addition, the expression of inducible nitric oxide synthase, nitric oxide, cyclooxygenase-2, pro-inflammatory cytokines (e.g. tumour necrosis factor-α and interleukin-1ß), and nuclear factor kappa-B in the epidermis were reduced after treating with surfactin-oleogel. Moreover, total cholesterol and free fatty acids were decreased, whereas the treatment of surfactin-oleogel increased triglycerides and linoleic acid content. Besides, immunohistochemical assay and real-time PCR analysis indicated that surfactin-oleogel blocked the TLR2-mediated NF-κB signalling pathways in the epidermis. Consequently, our results demonstrated that surfactin-oleogel had antibacterial and anti-inflammation activities to treat P. acnes-induced inflammatory acne vulgaris.Key points• Surfactin-oleogel effectively relieves inflammation and oxidative stress caused by P. acnes.• Surfactin-oleogel effectively reduced the P. acnes colony.• Surfactin-oleogel relieves P. acnes-induced inflammation by inactivated the TLR-mediated NF-κB.

Bacillomycin D-C16 inhibits growth of Fusarium verticillioides and production of fumonisin B1 in maize kernels.

Fusarium verticillioides causes ear and kernel rot in maize and produces mycotoxins, like fumonisin B1 (FB1). Bacillomycin D-C16 is a natural antimicrobial lipopeptide produced by Bacillus subtilis. In this study, the inhibitory effects of Bacillomycin D-C16 on the growth of F. verticillioides and on the production of FB1 in maize were investigated. Bacillomycin D-C16 displayed strong fungicidal activity against F. verticillioides, with a minimum inhibitory concentration (MIC) of 32 g/L. Scanning electron microscopy (SEM) showed that Bacillomycin D-C16 altered the morphology of F. verticillioides mycelia. Bacillomycin D-C16 reduced the ergosterol content, increased the release of nucleic acids and proteins, and increased the levels of reactive oxygen species (ROS) in fungal mycelia. Bacillomycin D-C16 also significantly inhibited the production of FB1 by inhibiting mycelial growth and decreasing the levels of fumonisin biosynthetic genes 1 (fum1), fum6 and fum14. The application of Bacillomycin D-C16 on maize kernels prior to storage inhibited the growth of F. verticillioides and the production of FB1. Our results suggested that Bacillomycin D-C16 has a significant antifungal activity that could be used as a potential natural antimicrobial agent to control food contamination and to ensure food safety.

Surfactin Mitigates a High-Fat Diet and Streptozotocin-Induced Type 2 Diabetes through Improving Pancreatic Dysfunction and Inhibiting Inflammatory Response.

Surfactin from Bacillus amyloliquefaciens fmb50 was utilized to treat mice with type 2 diabetes (T2DM) induced by a high-fat diet/streptozotocin (HFD/STZ). Our group's earlier research indicated that surfactin could lower blood glucose and mitigate liver dysfunction to further improve HFD/STZ-induced T2DM through modulating intestinal microbiota. Thus, we further investigated the effects of surfactin on the pancreas and colon in mice with T2DM to elucidate the detailed mechanism. In the present study, mice with HFD/STZ-induced T2DM had their pancreatic and colon inflammation, oxidative stress, and endoplasmic reticulum stress (ERS) reduced when given oral surfactin at a dose of 80 mg/kg body weight. According to further research, surfactin also improved glucose metabolism by activating the phosphatidylinositol kinase (PI3K)/protein kinase B (Akt) signaling pathway, further protecting islets ß-cell, promoting insulin secretion, inhibiting glucagon release and mitigating pancreas dysfunction. Additionally, after surfactin treatment, the colon levels of the tight junction proteins Occludin and Claudin-1 of T2DM mice were considerably increased by 130.64% and by 36.40%, respectively. These findings revealed that surfactin not only ameliorated HFD/STZ-induced pancreas inflammation and dysfunction and preserved intestinal barrier dysfunction and gut microbiota homeostasis but also enhanced insulin sensitivity and glucose homeostasis in T2DM mice. Finally, in the further experiment, we were able to demonstrate that early surfactin intervention might delay the development of T2DM caused by HFD/STZ, according to critical biochemical parameters in serum.

Combating biofilms of foodborne pathogens with bacteriocins by lactic acid bacteria in the food industry.

Most foodborne pathogens have biofilm-forming capacity and prefer to grow in the form of biofilms. Presence of biofilms on food contact surfaces can lead to persistence of pathogens and the recurrent cross-contamination of food products, resulting in serious problems associated with food safety and economic losses. Resistance of biofilm cells to conventional sanitizers urges the development of natural alternatives to effectively inhibit biofilm formation and eradicate preformed biofilms. Lactic acid bacteria (LAB) produce bacteriocins which are ribosomally synthesized antimicrobial peptides, providing a great source of nature antimicrobials with the advantages of green and safe properties. Studies on biofilm control by newly identified bacteriocins are increasing, targeting primarily onListeria monocytogenes, Staphylococcus aureus, Salmonella, and Escherichia coli. This review systematically complies and assesses the antibiofilm property of LAB bacteriocins in controlling foodborne bacterial-biofilms on food contact surfaces. The bacteriocin-producing LAB genera/species, test method (inhibition and eradication), activity spectrum and surfaces are discussed, and the antibiofilm mechanisms are also argued. The findings indicate that bacteriocins can effectively inhibit biofilm formation in a dose-dependent manner, but are difficult to disrupt preformed biofilms. Synergistic combination with other antimicrobials, incorporation in nanoconjugates and implementation of bioengineering can help to strengthen their antibiofilm activity. This review provides an overview of the potential and application of LAB bacteriocins in combating bacterial biofilms in food processing environments, assisting in the development and widespread use of bacteriocin as a promising antibiofilm-agent in food industries.

A mini-review: mechanism of antimicrobial action and application of surfactin.

Surfactin, an antibacterial lipopeptide produced by different strains of Bacillus subtilis, is a powerful biosurfactant. It also has multiple biological activities including antiviral, anti-mycoplasma and antiprotozoal activities, in addition to the broad-spectrum antimicrobial activities against Gram-positive bacteria, Gram-negative bacteria and fungi. Surfactin may be one of the promising alternatives to antibiotics. Surfactin's chemical structure and physicochemical properties are briefly discussed in this mini-review. Surfactin's antibacterial mechanism is mainly outlined as follows: (1) attacking pathogenic bacteria's cell membrane, causing cell membrane disintegration or osmotic pressure imbalance; (2) inhibiting pathogenic bacteria's protein synthesis, preventing cell reproduction; (3) inhibiting pathogenic bacteria's enzyme activity, affecting normal cell metabolism. This provides basis for the further research and application of surfactin. Finally, the application of surfactin in food and its prospect are summarized in brief.

Promoted Spore Formation of Bacillus amyloliquefaciens fmbJ by its Secondary Metabolite Bacillomycin D Coordinated with Mn2.

In Bacillus, the spore formation process is associated with the synthesis and release of secondary metabolites. A large number of studies have been conducted to systematically elucidate the pathways and mechanisms of spore formation. However, there are no studies have explored the relationship between secondary metabolites and spores. In this study, we investigated the relationship between its secondary metabolite bacillomycin D (BD) and spores using the simpler dipicolonic acid fluorimetry assay for spore counting in Bacillus amyloliquefaciens fmbJ. Our results showed that BD could promote the spore formation of B. amyloliquefaciens fmbJ and had a synergistic effect with certain concentrations of Mn2+. When 15.6 mg/L of BD and 1 mM of Mn2+ were added, the number of fmbJ spores increased from 1.42 × 108 CFU/mL to 2.02 × 108 CFU/mL after 36 h of incubation. The expressions of spore formation (kinA, kinB, kinC, kinD, kinE and spo0A) and Mn-related genes (mntA, mntH, mneS, mneP) were studied by RT-PCR. The results indicated that BD and Mn2+ promoted the spore formation of fmbJ by stimulating the transcription of kinB, kinD and increasing the influence of spo0F-spo0A phosphorylation transmission. This study provided a new idea to improve the spore production of B. amyloliquefaciens and laid the foundation for its industrial production. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01026-9.

Acetate Activates Lactobacillus Bacteriocin Synthesis by Controlling Quorum Sensing.

Bacteriocins are useful for controlling the composition of microorganisms in fermented food. Bacteriocin synthesis is regulated by quorum sensing mediated by autoinducing peptides. In addition, short-chain fatty acids, especially acetic acid, reportedly regulate bacteriocin synthesis. Five histidine kinases that regulated the synthesis of bacteriocins were selected to verify their interactions with acetate. Acetate activated the kinase activity of PlnB, SppK, and HpK3 in vitro and increased the yield of their cognate bacteriocins plantaricin EF, sakacin A, and rhamnosin B in vivo. The antimicrobial activity against Staphylococcus aureus of the fermentation supernatants of Lactobacillus plantarum, Lactobacillus sakei, and Lactobacillus rhamnosus with addition of acetate increased to 298%, 198%, and 289%, respectively, compared with that in the absence of acetate. Our study elucidated the activation activity of acetate in bacteriocin synthesis, and it might provide a potential strategy to increase the production of bacteriocin produced by Lactobacillus. IMPORTANCE Bacteriocins produced by lactic acid bacteria (LAB) are particularly useful in food preservation and food safety. Bacteriocins might increase bacterial competitive advantage against the indigenous microbiota of the intestines; at the same time, bacteriocins could limit the growth of undesired microorganisms in yogurt and other dairy products. This study confirmed that three kinds of histidine kinases were activated by acetate and upregulated bacteriocin synthesis both in vitro and in vivo. The increasing yield of bacteriocins reduced the number of pathogens and increased the number of probiotics in milk. Bacteriocin synthesis activation by acetate may have a broad application in the preservation of dairy products and forage silage.

Broccoli microgreens juice reduces body weight by enhancing insulin sensitivity and modulating gut microbiota in high-fat diet-induced C57BL/6J obese mice.

PURPOSE: This study aimed to explore the protective effect of broccoli microgreens juice (BMJ) during C57BL/6J mice obesity development. METHODS: The obese model mice, induced by feeding high-fat diet (HFD), were treated with BMJ by gavage for 10 weeks. Melbine was gavaged at 300 mg/(kg bw)/d, as a positive control group. RESULTS: BMJ supplementation significantly reduced white adipose tissues (WAT) mass, the body weight and adipocyte size, and increased water intake in HFD-fed mice. Moreover, it improved glucose tolerance, reduced insulin level and HOMA-IR value, and alleviated insulin resistance. Compared with the HFD group, BMJ supplementation significantly increased the relative abundance of Bacteroidetes and decreased the ratio of Firmicutes to Bacteroidetes at the phylum level, and enriched Bacteroides_acidifaciens at the species level. These changes in the composition of gut microbiota are associated with the production of short-chain fatty acids (SCFAs), and reduced LPS levels, and had an obvious anti-inflammatory effect. CONCLUSIONS: These findings suggested that the protective effects of BMJ on diet-induced obesity may be involved in gut microbiota-SCFAs-LPS-inflammatory axis. In addition, BMJ can enhance liver antioxidant capacity and reduce liver fat accumulation. Consequently, these results sustain BMJ as a novel functional food for obesity, on the basis of its opposing effects on HFD-induced obesity in mice.

The determination of antibacterial mode for cationic lipopeptides brevibacillins against Salmonella typhimurium by quantum chemistry calculation.

Brevibacillins are broad-spectrum cationic antimicrobial lipopeptides produced by Brevibacillus laterosporus fmb70 CGMCC 18426. The antibacterial mode of brevibacillins against Salmonella typhimurium CICC 21493 was investigated by quantum chemistry calculation in this study. The addition of LPS, Mg2+, and Ca2+ partially reduced the antimicrobial activity of brevibacillin and brevibacillin V against S. typhimurium, which indicated that the two cationic lipopeptides could bind to LPS and displaced the divalent cations on the LPS network. Release of LPS from S. typhimurium by brevibacillin and brevibacillin V resulted in destroying the dense LPS network and increasing the permeability of the outer membrane. Quantum chemistry calculation analysis revealed that Lys7 is the most critical amino acid residue to destroy the outer membrane. The total average N-H charge difference of the three protonated amino groups (Orn3-NH3, Lys7-NH3, and Lys10-NH3) determined the ability of brevibacillin V to bind LPS stronger than brevibacillin. Calcein complete leakage from liposomes and release of DiSC3-5 from the cytoplasmic membrane (CM) indicated that brevibacillin and brevibacillin V may destroy the CM. Brevibacillin and brevibacillin V exhibited their antimicrobial activities through membrane damages, where the OM permeability with high concentration of 64-256 µg/mL and membrane damage of CM with a low concentration of 4 µg/mL. Our finding might be helpful to understand the broad-spectrum antimicrobial mechanism of cationic lipopeptide and to design the novel antimicrobial peptide. KEY POINTS: • Brevibacillin V had stronger affinity for LPS than brevibacillin. • The N-H charge difference was the key of the difference in the affinity to LPS. • Brevibacillins inhibited Salmonella by displacing the divalent cations on the LPS.

Maltose effective improving production and regulatory biosynthesis of plantaricin EF in Lactobacillus plantarum 163.

Plantaricin EF, a kind of natural antibacterial substance, has shown inhibitory effect on most pathogen and spoilage microorganisms, which possessed great potential in food preservation. However, the lower production of plantaricin EF has limited its large-scale production and application. In this study, the effect of maltose on plantaricin EF production and its regulation mechanism in Lactobacillus plantarum 163 were investigated. Maltose significantly improved the biomass and plantaricin EF production, which increased by 3.35 and 3.99 times comparing to the control without maltose, respectively. The maximum production of plantaricin E and F in fed-batch fermentation were 10.55 mg/L and 22.94 mg/L, respectively. Besides, qPCR results showed that maltose remarkably improved transcription of plnA, plnB, plnD, plnE, plnF, plnG1 and plnH, and heighten transcription of lamR, lamK, hpk6 and rrp6. These results provided an effective method to enhance plantaricin EF production and revealed a possible regulatory mechanism from transcriptome results that hpk6, rrp6, lamK and lamR were relative to plantaricin EF production. Genes, hpk6 and rrp6, promote transcription of plnG1, whereas lamK and lamR enhance transcription of plnA, plnB and plnD, which increased plantaricin EF production. KEYPOINTS: • Maltose was proved to be effective in promoting the biosynthesis of plantaricin EF. • Maltose promoted the transcription of biosynthesis and secretion genes of plantaricin EF. • Up-regulation of genes lamR, lamK, hpk6 and rrp6 heightened the plantaricin EF production.

Preharvest UVB Application Increases Glucosinolate Contents and Enhances Postharvest Quality of Broccoli Microgreens.

Broccoli microgreens have shown potential health benefits due to their high glucosinolate (GL) levels. Previously, we observed that postharvest UVB treatment did not have much effect on increasing GLs in broccoli microgreens. In this study, we investigated the influence of preharvest UVB irradiation on GL levels in broccoli microgreens. UHPLC-ESI/ITMS analysis showed that preharvest UVB treatments with UVB 0.09 and 0.27 Wh/m2 significantly increased the glucoraphanin (GLR), glucoerucin (GLE), and total aliphatic GL levels by 13.7 and 16.9%, respectively, in broccoli microgreens when measured on harvest day. The nutritional qualities of UVB-treated microgreens were stable during 21-day storage, with only small changes in their GL levels. Broccoli microgreens treated before harvest with UVB 0.27 Wh/m2 and 10 mM CaCl2 spray maintained their overall quality, and had the lowest tissue electrolyte leakage and off-odor values during the storage. Furthermore, preharvest UVB 0.27 Wh/m2 treatment significantly increased GL biosynthesis genes when evaluated before harvest, and reduced the expression level of myrosinase, a gene responsible for GL breakdown during postharvest storage. Overall, preharvest UVB treatment, together with calcium chloride spray, can increase and maintain health-beneficial compound levels such as GLs and prolong the postharvest quality of broccoli microgreens.

Iturin A Induces Resistance and Improves the Quality and Safety of Harvested Cherry Tomato.

The soft rot disease caused by Rhizopus stolonifer is an important disease in cherry tomato fruit. In this study, the effect of iturin A on soft rot of cherry tomato and its influence on the storage quality of cherry tomato fruit were investigated. The results showed that 512 µg/mL of iturin A could effectively inhibit the incidence of soft rot of cherry tomato fruit. It was found that iturin A could induce the activity of resistance-related enzymes including phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), glucanase (GLU), and chitinase (CHI), and active oxygen-related enzymes including ascorbate peroxidases (APX), superoxide dismutases (SOD), catalases (CAT), and glutathione reductase (GR) of cherry tomato fruit. In addition, iturin A treatment could slow down the weight loss of cherry tomato and soften the fruit. These results indicated that iturin A could retard the decay and improve the quality of cherry tomato fruit by both the inhibition growth of R. stolonifera and the inducing the resistance.

Genome Mining, Heterologous Expression, Antibacterial and Antioxidant Activities of Lipoamides and Amicoumacins from Compost-Associated Bacillus subtilis fmb60.

Bacillus subtilis fmb60, which has broad-spectrum antimicrobial activities, was isolated from plant straw compost. A hybrid NRPS/PKS cluster was screened from the genome. Sixteen secondary metabolites produced by the gene cluster were isolated and identified using LC-HRMS and NMR. Three lipoamides D-F (1-3) and two amicoumacin derivatives, amicoumacins D, E (4, 5), were identified, and are reported here for the first time. Lipoamides D-F exhibited strong antibacterial activities against harmful foodborne bacteria, with the MIC ranging from 6.25 to 25 µg/mL. Amicoumacin E scavenged 38.8% of ABTS+ radicals at 1 mg/mL. Direct cloning and heterologous expression of the NRPS/PKS and ace gene cluster identified its importance for the biosynthesis of amicoumacins. This study demonstrated that there is a high potential for biocontrol utilization of B. subtilis fmb60, and genome mining for clusters of secondary metabolites of B. subtilis fmb60 has revealed a greater biosynthetic potential for the production of novel natural products than previously anticipated.