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Background Probiotics are defined as microorganisms that can exert health benefits for the host. Among the recognized probiotics, Lactobacillus paracasei are one of the most frequently used probiotics in humans. The L. paracasei strain M11-4, isolated from fermented rice, which could ferment soymilk within a short curd time, and fermented soymilk presented high viability, acceptable flavor and antioxidant activity, which revealed that the strain maybe have a potential antioxidant value. Therefore, it is necessary to further explore the antioxidant activity of L. paracasei strain M11-4. Results The radical scavenging activities, lipid peroxidation inhibition, and reducing power of L. paracasei M11-4 were the highest in the fermentation culture without cells, while the activities of other antioxidant enzymes of L. paracasei M11-4 were high in the cell-free extract and bacterial suspension. Moreover, L. paracasei M11-4 exerted its antioxidant effect by upregulating the gene expression of its antioxidant enzymes, thioredoxin (Trx) and glutathione (GSH) systems when hydrogen peroxide (H2 O2 ) existed. Supplementation of rats with L. paracasei M11-4 effectively alleviated D-galactose-induced oxidative damage in the liver and serum and prevented D-galactose-induced changes to intestinal microbiota. Supplementation with L. paracasei M11-4 also reduced the elevated expression of Trx and GSH system genes induced by D-galactose. Conclusion L. paracasei M11-4 has good antioxidant properties both in vitro and in vivo and the antioxidant mechanism of it was studied at the molecular level. This article is protected by copyright. All rights reserved.

The quorum-sensing regulation of class II bacteriocin (AcH) synthesis in Lactobacillus plantarum subsp. plantarum Zhang-LL was studied. No detectable inhibition zone was formed by the supernatant of L. plantarum subsp. plantarum Zhang-LL culture in skim milk (SM) with an inoculum size of 7 × 102 CFU/mL after incubation for 36 h. Hence, this culture system was used to investigate the induced regulation mechanism of bacteriocin production in L. plantarum subsp. plantarum Zhang-LL. Bacteriocin production by this bacterium in SM medium was induced by treatment with inactivated culture supernatant from de Man Rogosa Sharpe (MRS) medium (supernatant-MRS). Pediocin AcH encoded by the papA gene in a plasmid in strain Zhang-LL was the inducer present in supernatant-MRS. This is the first report of the role of pediocin AcH in the quorum-sensing regulation of class II bacteriocin synthesis. The mRNA of the papA, papB, papC, and papD genes involved in bacteriocin synthesis by strain Zhang-LL in SM medium was upregulated significantly after being induced by pediocin AcH. This study offers the first evidence that the ABT40_05745, ABT40_05750, and ABT40_11975 components of two-component systems in L. plantarum subsp. plantarum Zhang-LL are involved in the induced regulation of AcH bacteriocin production. HIGHLIGHTS

Plantaricin BM-1, a class IIa bacteriocin produced by Lactobacillus plantarum BM-1, shows obvious antibacterial activity against Escherichia coli. However, the mechanism underlying the action of class IIa bacteriocins against gram-negative bacteria remains to be explored. The purpose of this study was to investigate the role of YbfA, a DUF2517 domain-containing protein, in the response of Escherichia coli K12 to plantaricin BM-1. The growth curve experiment and MIC experiment showed that the sensitivity of E. coli to plantaricin BM-1 was decreased by a ybfA null mutation. Electron microscopy showed that the ybfA null mutation reduced the surface rupture and contraction caused by plantaricin BM-1, and mitigated the effect of plantaricin BM-1 on the morphology of the E. coli cell membrane. Proteomics analysis showed that 323 proteins were differentially expressed in E. coli lacking the ybfA gene (P < 0.05); 118 proteins were downregulated, and 205 proteins were upregulated. The metabolic pathways containing the upregulated proteins mainly included outer membrane proteins, integral components of the plasma membrane, regulation of cell motility, and regulation of locomotion. The metabolic pathways involving the downregulated proteins mainly included outer membrane protein glycine betaine transport, amino-acid betaine transport, and transmembrane signaling receptor activity. The results of the proteomics analysis showed that the protein expression of the BasS/BasR two-component system was significantly increased (P < 0.05). Moreover, the expression levels of downstream proteins regulated by this two-component system were also significantly increased, including DgkA, FliC, and MlaE, which are involved in cell membrane structure and function, and RT-qPCR also confirmed this result. The growth curve showed that the sensitivity of E. coli to plantaricin BM-1 was significantly increased due to deletion of the BasS/BasR two-component system. Thus, deletion of ybfA in E. coli can increase the expression of the BasS/BasR two-component system and positively regulate the structure and function of the cell membrane to reduce the sensitivity to plantaricin BM-1. This will help to explore the mechanism of action of class IIa bacteriocins against gram-negative bacteria.

Probiotics can improve dyslipidemia and promote metabolic control as a therapeutic approach for type 2 diabetes mellitus (T2DM). The hypoglycemic effects of space-induced Lactobacillus plantarum SS18-5 on T2DM were explored in 4-week-old male Sprague Dawley rats. The normal (N) group was fed a basal diet, while the other groups received a high glucose fat diet. T2DM was established by streptozotocin injection and the T2DM rats were randomly divided into three groups, a diabetic (D) group (T2DM rats treated with saline only), GS18 group (T2DM rats treated with 109 CFU/ml of L. plantarum GS18), and SS18-5 group (T2DM rats treated with 109 CFU/ml of L. plantarum SS18-5). After continuous gavage for 6 weeks, blood biochemical indices were measured and livers were collected for histopathological examination. The colon contents were collected for counting of Escherichia coli, Clostridium perfringens, and Lactobacillus sp. The results showed that L. plantarum SS18-5 effectively controlled the weight of rats, reduced levels of fasting blood glucose, glycosylated hemoglobin, and insulin, increased liver glycogen levels, improved abnormal metabolism of blood lipids, enhanced the effect of anti-lipid peroxidation, alleviated chronic inflammation and fatty liver disease, and regulated the intestinal microbiota by reducing the numbers of E. coli and C. perfringens, and increasing the numbers of Lactobacillus sp. From these results, we conclude that space-induced L. plantarum SS18-5 has the potential to improve T2DM by alleviating hypoglycemia and regulating the intestinal microbiota. PRACTICAL APPLICATIONS: With the exploration of the universe, a large number of studies have observed the changes of microorganisms in space flight, which provided a new method for high-quality microbial pharmaceuticals in the space environment. In this study, the space environment mutated. Lactobacillus plantarum SS18-5 can effectively improve the blood glucose of rats with type 2 diabetes, relieve oxidative stress, reduce blood lipid content, enhance immune capacity, and regulate intestinal microflora, which has potential use in the treatment of type 2 diabetes.

In this study, an active antibacterial packaging film was developed by coating a polyethylene terephthalate/polyvinylidene chloride/retort casting polypropylene (PPR) plastic multilayer film with plantaricin BM‐1 and chitosan. The characteristics of the active packaging film and its antibacterial effect for chilled meat preservation were evaluated. Our results indicated that the barrier properties against oxygen were improved significantly and the tensile strength and the elongation at break were changed slightly. The active plantaricin film significantly (P

Bacillus subtilis SH21 was observed to produce an antifungal protein that inhibited the growth of F. solani. To purify this protein, ammonium sulfate precipitation, gel filtration chromatography, and ion-exchange chromatography were used. The purity of the purified product was 91.33% according to high-performance liquid chromatography results. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis revealed that the molecular weight of the protein is 30.72 kDa. The results of the LC–MS/MS analysis and a subsequent sequence-database search indicated that this protein was a chitosanase, and thus, we named it chitosanase SH21. Scanning and transmission electron microscopy revealed that chitosanase SH21 appeared to inhibit the growth of F. solani by causing hyphal ablation, distortion, or abnormalities, and cell-wall depression. The minimum inhibitory concentration of chitosanase SH21 against F. solani was 68 µg/mL. Subsequently, the corresponding gene was cloned and sequenced, and sequence analysis indicated an open reading frame of 831 bp. The predicted secondary structure indicated that chitosanase SH21 has a typical a-helix from the glycoside hydrolase (GH) 46 family. The tertiary structure shared 40% similarity with that of Streptomyces sp. N174. This study provides a theoretical basis for a topical cream against fungal infections in agriculture and a selection marker on fungi.

Plantaricin BM-1, a class IIa bacteriocin produced by Lactobacillus plantarum BM-1, has significant antibacterial activity against Gram-positive and Gram-negative bacteria. This study aimed to explore the role of the Escherichia coli K12 outer membrane (OM) channel protein TolC in the response to plantaricin BM-1. The tolC null mutant (E. coli K12∆tolC) was constructed by Red homologous recombination. The mechanism of tolC regulating the sensitivity of E. coli K12 under plantaricin BM-1 was investigated. tolC null mutation significantly increased the E. coli K12 sensitivity to plantaricin BM-1 and inhibited biofilm formation, and cells ruptured and shrunk. Proteomic analysis showed that the AcrAB-TolC and EmrAB-TolC efflux pumps were significantly (p

Plantaricin BM-1 is a class IIa bacteriocin with a strong bactericidal effect on gram-positive bacteria. Although plantaricin BM-1 also inhibits the growth of some gram-negative bacteria, including Escherichia coli, the mechanism is not clear. In this study, we used tandem mass tag-based quantitative proteomics analysis to examine the inhibitory mechanism of plantaricin BM-1 against E. coli K12, and evaluated the morphological effects by electron microscopy. The results demonstrated that plantaricin BM-1 inhibits the growth of E. coli K12 by bacteriostatic action, mainly acting on the surface of the cell wall, leading to its collapse. Proteomic analysis identified 976 differentially expressed proteins (>1.2-fold change, p < 0.05) under treatment with plantaricin BM-1, including 490 up-regulated proteins and 486 down-regulated proteins. These proteins were mainly involved in peptidoglycan synthesis and energy metabolism pathways, including amino acid, glyoxylate and dicarboxylate, ABC transporter, and quorum-sensing pathways. Specifically, plantaricin BM-1 treatment significantly improved peptidoglycan synthesis and enhanced the tricarboxylic acid cycle in E. coli K12, and altered the expression of cell membrane proteins. These results provide new insight into the inhibition mechanism of class IIa bacteriocins on gram-negative bacteria, which can lay the foundation for its broader use as an alternative to conventional antibiotics.

AIMS The aim of this study was to develop a plantaricin BM-1, a typical IIa bacteriocin produced by Lactocacillus plantarumBM-1, for active polyvinylidene chloride (PVDC) films and to determine the antimicrobial effect of plantaricin BM-1 incorporated into a PVDC film on fresh pork during 7 days of storage at 4°C. METHODS AND RESULTS Plantaricin BM-1 solutions (20 480 AU ml-1 ) that absorbed into the PVDC film increased gradually and reached maximum volumes during exposure for up to 20 h. When soaked in water, the released amount of plantaricin BM-1 from the active PVDC film reached a maximum at 20 h. The plantaricin BM-1 active PVDC film had an obvious antilisterial effect in culture medium and fresh pork inoculated with Listeria monocytogenes. Furthermore, plantaricin BM-1-incorporated PVDC film was also significantly (P

Plantaricin BM-1, a typical IIa bacteriocin, is produced by Lactobacillus plantarum BM-1, which can be isolated from a traditionally fermented Chinese meat product. This bacteriocin exhibits perfect thermal stability and broad inhibitory activity against certain foodborne pathogens. In this study, we investigated the effect of plantaricin BM-1 combined with physicochemical treatments (sodium nitrite, heat treatment, ultrahigh pressure technology) on the control of Listeria monocytogenes populations in cooked ham. According to our results, the addition of sodium nitrite (0.075 or 0.15 mg/mL) significantly inhibited the growth of L. monocytogenes in broth, plantaricin BM-1 combined with heat treatment reduced the viable counts of L. monocytogenes more than plantaricin BM-1 alone, and plantaricin BM-1 combined with sodium nitrite and heat treatment had the greatest antibacterial effect in broth. However, the addition of sodium nitrite or heat treatment alone did not inhibit the growth of L. monocytogenes in cooked ham. The combined application of 5,120 arbitrary units per gram (AU/g) plantaricin BM-1 and 0.075 mg/g sodium nitrite reduced L. monocytogenes significantly more effectively (by 1.97 log CFU/g) than application of 5,120 AU/g plantaricin alone. Plantaricin BM-1 application combined with sodium nitrite and heat treatment led to viable counts of L. monocytogenes below the level of detection for 49 days of storage. Moreover, in cooked ham treated with 5,120 AU/g plantaricin BM-1, 0.075 mg/g sodium nitrite, and ultrahigh pressure technology (400 MPa for 5 min), viable counts of L. monocytogenes were reduced by 5.79 log CFU/g compared with control samples at the end of storage. In conclusion, the combination of plantaricin BM-1 with physicochemical treatments had a synergistic inhibitory effect on L. monocytogenes in cooked ham.

Background Probiotics are defined as microorganisms that can exert health benefits for the host. Among the recognized probiotics, Bifidobacterium are the most frequently used probiotics in humans. The aim of this study was to evaluate the antidiabetic activity of Bifidobacterium strains isolated from breastfed infant faeces, both in vitro, using the Caco-2 monolayer transwell model, and in vivo, using a mice model of impaired glucose tolerance induced by a high-fat diet (HFD). Results The cell-free supernatant of Bifidobacterium lactis A12 showed better inhibitory activity of α-glucosidase and inhibited the glucose absorption and transport than B. lactis BB12, which is a typical probiotic with antidiabetic capabilities. B. lactis A12 improved the impaired glucose intolerance, restored islet function and morphology with insulin resistance induced by the HFD in C57BL/6J mice. Furthermore, in small intestine tissues, the cell-free supernatant of B. lactis A12 decreased the messenger RNA expressions of sucrase-isomaltase, live B. lactis A12 cells decreased glucose transporters 2. B. lactis A12 significantly stimulated the glucagon like peptide-1 (GLP-1) secretion and upregulated proglucagon messenger RNA levels. Conclusion B. lactis A12 protect against the deleterious effects of HFD-induced diabetes by inhibiting the utilization, absorption, and transport of glucose by intestinal epithelial cells and promoting the expression and secretion of GLP-1. © 2020 Society of Chemical Industry.

Escherichia colican induce a group of stress-response proteins, including carbon starvation-induced lipoprotein (Slp), which is an outer membrane lipoprotein expressed in response to stressful environments. In this paper,slpnull mutantE. coliwere constructed by insertion of the group II intron, and then the growth sensitivity of theslpmutant strain was measured under 0.6% (vol/vol) hydrogen peroxide. The changes in resistance to hydrogen peroxide stress were investigated by detecting antioxidant activity and gene expression in theslpmutant strain. The results showed that deletion of theslpgene increased the sensitivity ofE. coliunder 0.6% (vol/vol) hydrogen peroxide oxidative stress. Analysis of the unique mapping rates from the transcriptome libraries revealed that four of thirteen remarkably up/down-regulated genes inE. coliwere involved in antioxidant enzymes after mutation of theslpgene. Mutation of theslpgene caused a significant increase in catalase activity, which contributed to an increase in glutathione peroxidase activity. ThekatGgene was activated by the OxyR regulator, which was activated directly by 0.6% (vol/vol) hydrogen peroxide, and HPI encoded bykatGwas induced against oxidative stress. Therefore, the carbon starvation-induced lipoprotein Slp regulates the expression of antioxidant enzymes and the transcriptional activator OxyR in response to the hydrogen peroxide environment, ensuring that cells are protected from hydrogen peroxide oxidative stress at the level of enzyme activity and gene expression.

To investigate the effects of Kluyveromyces marxianus M3 isolated from Tibetan mushrooms on diet-induced hypercholesterolemia in rats, female Wistar rats were fed a high-cholesterol diet (HCD) for 28 d to generate hyperlipidemic models. Hyperlipidemic rats were assigned to four groups, which were individually treated with three different dosages of K. marxianus M3+HCD or physiological saline+HCD via oral gavage for 28 d. The total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels in the serum and liver of the rats were measured using commercially available enzyme kits. In addition, the liver morphology was also examined using hematoxylin and eosin staining and optical microscopy. According to our results, the serum and liver TC, TG, LDL-C levels and atherogenic index (AI) were significantly decreased in rats orally administered K. marxianus M3 (p

In this study, the regularity of autoinducer-2 (AI-2) secretion during growth and the effect of the addition of various carbohydrates on AI-2 secretion in Bifidobacterium longum subsp. longum BBMN68 were investigated. The results indicated that the AI-2 concentration reached its highest level (2536·60 nmol l-1 ) at the early stationary growth phase, and then decreased to 1263·72 nmol l-1 at the late stationary growth phase in Bifidobacterium cultures. When the density of the cultures which mannose, fructose, sucrose and lactose had been added to reached an OD600 nm of 1·0, the AI-2 concentrations in the cultures were 1953·84, 1637·34, 1200·99 and 1077·60 nmol l-1 , respectively. These concentrations were all significantly higher than that of the control culture (1031·33 nmol l-1 ). Similarly, the addition of fructooligosaccharide significantly increased the AI-2 concentrations to 2094·29 nmol l-1 . This study provides the advanced evidence that certain carbohydrates promote the secretion of AI-2, and that this occurs at the single cell level and is therefore unaffected by cell density. Significance and impact of the study This study provided the advanced data of the regularity of autoinducer-2 (AI-2) secretion during growth and the promotion on AI-2 secretion of different added carbohydrates in Bifidobacterium, which may be a new potential strategy to improve the acid resistance of Bifidobacterium applied in the food industry.

Inducing at sublethal pHs can strengthen the survival rate of Bifidobacterium at the lethal pH. In this study, we determined the optimal inducing condition of Bifidobacterium and analyzed the effect of a low pH environment on fatty acid (FA) composition and thiol content in Bifidobacterium. The result indicates that inducing at pH 4.5 significantly improved the survival rate of Bifidobacterium at pH 3,5 and 3.0. After being induced at pH 4.5, the percentage of C18:1 increased significantly, while the percentage of C16:0 decreased significantly. The results suggest inducing treatment strengthened the membrane fluidity of Bifidobacterium. After being stressed at the lethal pH, the percentage of saturated FA increased significantly in both the control and induced cells, while the percentage of unsaturated FA decreased significantly. The results suggested that, to survive at the lethal pH, the cells strengthened the membrane rigidity and weakened the membrane fluidity to block the entrance of H+ by changing the FA composition in Bifidobacterium. In addition, after being stressed, the thiol content of the control cells was significantly lower than that of the induced cells. This study provided the first data proof that showed FA and thiol participated in response to low pH in Bifidobacterium.

The effects of the Kluyveromyces marxianus M3 strain, isolated from Tibetan mushrooms, on plasma lipids, egg cholesterol level, egg quality, and intestinal health of laying hens were evaluated. In total, 160 Beijing fatty laying hens (43 weeks old) were divided into four groups and fed a basal diet supplemented with 0%, 0.1%, 0.3%, or 0.5% freeze-dried K. marxianus M3 powder for four weeks. The results showed that yeast supplementation reduced serum total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), and very low density lipoprotein-cholesterol (VLDL-C) levels (p

Two application methods were used to investigate the effect of plantaricin BM-1 on the control of Listeria monocytogenes and background spoilage bacteria in sliced vacuum-packaged cooked ham without the addition of any chemical preservatives, including sodium nitrite, during 35 days of storage at 4°C. Regardless of the application method, plantaricin BM-1 treatment (320, 640, or 1,280 arbitrary units [AU]/g of sliced cooked ham) significantly (P < 0.05) reduced the survival of L. monocytogenes (inoculated at 4 log CFU/g of sliced ham) compared with its survival in the control during the first 21 days of storage at 4°C. The inhibitory effect of plantaricin applied to the surface of the ham was significantly better than the same concentration of plantaricin incorporated into the cooked ham (P < 0.0001) during storage. Even 320 AU/g plantaricin applied to the surface exhibited greater inhibition of L. monocytogenes than 1,280 AU/g plantaricin incorporated into the cooked ham on days 1, 14, and 28. A level of 1,280 AU/g plantaricin applied to the surface of the ham reduced L. monocytogenes counts to below the detection limit from the 1st to the 21st day of storage at 4°C. Afterwards, L. monocytogenes was able to regrow, and the viable counts of L. monocytogenes at the end of storage reached 2.76 log CFU/g (6.11 log CFU/g lower than in the control). In the control ham, the counts of background spoilage bacteria increased gradually and surpassed the microbiological spoilage limitation level on the 21st day of storage. However, plantaricin BM-1 treatment significantly (P < 0.05) reduced the survival of background spoilage bacteria in ham compared with their survival in the control from day 21 to 35 of storage at 4°C. A level of 1,280 AU/g plantaricin incorporated into cooked ham was the most effective, reducing the count of background spoilage bacteria count from an initial 2.0 log CFU/g to 1.5 log CFU/g on day 7. This was then maintained for another 14 days and finally increased to 2.76 log CFU/g at the end of the storage at 4°C (2.85 log CFU/g lower than in the control). In conclusion, plantaricin BM-1 application inhibited the growth of L. monocytogenes and background spoilage bacteria in cooked ham during storage at 4°C and could be used as an antimicrobial additive for meat preservation.

The effect of plantaricin BM-1 on the behavior of Listeria Monocytogenes in sliced brined cooked ham during refrigerated storage (4°Cand 9°C) was assessed. The combination of low storage temperature (4°C) and plantaricin can significantly reduced viable counts of L. Monocytogenes during storage (P < 0.05). When the same amount of plantaricin was applied, the growth of L. Monocytogenes in brined cooked ham stored at 4 °C was significantly slower than that in sample stored at 9°C (P < 0.01). Even combination of 1280 AU/g plantaricin and 4°C storage produced greater inhibition of L. monocytogenes than combination of 5120 AU/g plantaricin and 9°C. The combination of 4°C and 5120 AU/g plantaricin was the most effective treatment and can decrease the levels of L. monocytogenes 1.7 Log CFU/g at the end of storage. Keywords-component; refrigerated storage; plantaricin; Listeria Monocytogenes; brined cooked ham

In recent years, bacteriocin as a natural antimicrobial compound, provides enormous promise to be used in food safety preservation. In this work, the polyethylene(PE)-based biological preservative films incorporating plantaricin BM-1, a typical IIa bacteriocin produced by Lactobacillus plantarum BM-1, were developed and characterized . The results showed that polyethylene (PE), low-density polyethylene (LDPE) and high-density polyethylene (HDPE) films soaked in plantaricin BM-1 solution had obvious antimicrobial activities aganist Listeria monocytogenes . And the volume of plantaricin BM-1 solution absorbed by PE, LDPE and HDPE films continued to increase and reached the maximum during exposure for up to 10, 6 and 16 hours, respectively. And the maximum absorption volumes of plantaricin BM-1 solution had no significant difference ( p >0.05) between the PE, LDPE and HDPE films. When soaking in water, the release amount of plantaricin BM-1 from active PE, LDPE and HDPE films reached the maximum potency at 16, 12 and 20 hours respectively. And the maximum release amount of plantaricin BM-1 from PE and LDPE active films were dramatically more than the HDPE active film ( p

Flowering peach trees, whose flowers have not be used and result in lot of waste of resources. This paper detected 18 kinds of amino acids and eight trace elements to provide data support for resource utilization of flowering peach blossom. Results showed that two amino acids (Asp and Pro) were higher than 2.0%, Glu, Lys, Leu, Ala and Val were more than 1.0% in flowers of flowering peach, especially Pro was the highest. Flowers of flowering peach contain many kinds of mineral elements, Mg, Ca, K and Mn were very high, which is beneficial for human health. All these data proved that flowers of flowering peach have high value as foods.