RESUMO
Plasmin-induced protein hydrolysis significantly compromises the stability of ultrahigh-temperature (UHT) milk. ß-Lactoglobulin (ß-Lg) was observed to inhibit plasmin activity, suggesting that there were active sites as plasmin inhibitors in ß-Lg. Herein, plasmin inhibitory peptides were explored from ß-Lg using experimental and computational techniques. The results revealed that increased denaturation of ß-Lg enhanced its affinity for plasmin, leading to a stronger inhibition of plasmin activity. Molecular dynamics simulations indicated that electrostatic and van der Waals forces were the primary binding forces in the ß-Lg/plasmin complex. Denatured ß-Lg increased hydrogen bonding and reduced the binding energy with plasmin. The sites of plasmin bound to ß-Lg were His624, Asp667, and Ser762. Four plasmin inhibitory peptides, QTMKGLDI, EKTKIPAV, TDYKKYLL, and CLVRTPEV, were identified from ß-Lg based on binding sites. These peptides effectively inhibited plasmin activity and enhanced the UHT milk stability. This study provided new insights into the development of novel plasmin inhibitors to improve the stability of UHT milk.
Assuntos
Fibrinolisina , Lactoglobulinas , Leite , Lactoglobulinas/química , Animais , Leite/química , Fibrinolisina/química , Fibrinolisina/metabolismo , Fibrinolisina/antagonistas & inibidores , Bovinos , Temperatura Alta , Armazenamento de Alimentos , Simulação de Dinâmica Molecular , Antifibrinolíticos/química , Peptídeos/química , Peptídeos/farmacologiaRESUMO
Lactic-acid-bacteria-derived bacteriocins are used as food biological preservatives widely. Little information is available on the impact of bacteriocin intake with food on gut microbiota in vivo. In this study, the effects of fermented milk supplemented with nisin (FM-nisin) or plantaricin Q7 (FM-Q7) from Lactiplantibacillus plantarum Q7 on inflammatory factors and the gut microbiota of mice were investigated. The results showed that FM-nisin or FM-Q7 up-regulated IFN-γ and down-regulated IL-17 and IL-12 in serum significantly. FM-nisin down-regulated TNF-α and IL-10 while FM-Q7 up-regulated them. The results of 16S rRNA gene sequence analysis suggested that the gut microbiome in mice was changed by FM-nisin or FM-Q7. The Firmicutes/Bacteroides ratio was reduced significantly in both groups. It was observed that the volume of Akkermansia_Muciniphila was significantly reduced whereas those of Lachnospiraceae and Ruminococcaceae were increased. The total number of short-chain fatty acids (SCFAs) in the mouse feces of the FM-nisin group and FM-Q7 group was increased. The content of acetic acid was increased while the butyric acid content was decreased significantly. These findings indicated that FM-nisin or FM-Q7 could stimulate the inflammation response and alter gut microbiota and metabolic components in mice. Further in-depth study is needed to determine the impact of FM-nisin or FM-Q7 on the host's health.
Assuntos
Microbioma Gastrointestinal , Lactobacillales , Nisina , Camundongos , Animais , Nisina/metabolismo , Nisina/farmacologia , Leite/metabolismo , RNA Ribossômico 16S/genética , Lactobacillales/metabolismo , Ácido ButíricoRESUMO
Bacteriocins are crucial metabolites of probiotics that display beneficial functions. The intestinal barrier is an important target on which probiotics exert their intestinal health activity. However, the impacts of bacteriocin-producing probiotics on the intestinal barrier are unclear. In this study, the effects of bacteriocin-producing Lactiplantibacillus plantarum Q7 and L. plantarum F3-2 on the intestinal barrier of mice were explored. It was shown that L. plantarum Q7 promoted the expression of mucin MUC2 to enhance the protection provided by the intestinal mucus layer. L. plantarum Q7 up-regulated the gene expression of intestinal tight junction proteins ZO-1 and JAM-1 significantly, and L. plantarum F3-2 up-regulated ZO-1 and Claudin-1 markedly, which exhibited tight junction intestinal barrier function. The two strains promoted the release of IgA and IgG at varying degrees. The antimicrobial peptide gene RegIIIγ was up-regulated markedly, and the gene expression of inflammatory cytokines appeared to exhibit an upward trend with L. plantarum Q7 treatment, so as to enhance intestinal immune regulation function. Furthermore, L. plantarum Q7 and L. plantarum F3-2 increased the abundance of the beneficial bacteria Muribaculaceae, inhibited the growth of the harmful bacteria Parabacteroides, and facilitated the synthesis of total short-chain fatty acids (SCFAs), which seemed to favor the prevention of metabolic diseases. Our results suggested that L. plantarum Q7 and L. plantarum F3-2 showed strain specificity in their protective effects on the intestinal chemical, physical, immunological and biological barriers of mice, which provided theoretical support for the selective utilization of bacteriocin-producing strains to regulate host health.
Assuntos
Bacteriocinas , Intestinos , Animais , Camundongos , Defecação , Peptídeos Antimicrobianos , Bacteriocinas/farmacologia , BacteroidetesRESUMO
Bacteriocins production is one of important beneficial characteristics of probiotics, which has antibacterial property against intestinal pathogens and is helpful for regulating intestinal flora. To investigate the impact of bacteriocin-producing probiotics on gut microecology, bacteriocin-producing Lactiplantibacillus plantarum YRL45 was orally administered to mice. The results revealed that it promoted the release of cytokines and improved the phagocytic activity of peritoneal macrophages to activate the immune regulation system. L. plantarum YRL45 was conducive to maintaining the morphology of colon tissue without inflammation and increasing the ratio of villus height to crypt depth in the ileum. The gene expression levels of Muc2, ZO-1 and JAM-1 were significantly up-regulated in the ileum and colon, and the gene expression of Cramp presented an upward trend with L. plantarum YRL45 intervention. Moreover, L. plantarum YRL45 remarkably enhanced the levels of immunoglobulins sIgA, IgA and IgG in the intestine of mice. The 16S rRNA gene analysis suggested that L. plantarum YRL45 administration up-regulated the relative abundance of the beneficial bacteria Muribaculaceae and Akkermansia, down-regulated the abundance of the pathogenic bacteria Lachnoclostridium, and promoted the production of acetic acid, propionic acid and total short-chain fatty acids (SCFAs) in mice feces. Our findings indicated that L. plantarum YRL45 had the potential to be developed as a novel probiotic to regulate the intestinal barrier by altering gut microbiota to enhance intestinal immunity and ameliorate intestinal flora balance.
Assuntos
Bacteriocinas , Microbioma Gastrointestinal , Lactobacillus plantarum , Probióticos , Animais , Camundongos , Bacteriocinas/farmacologia , Bacteriocinas/genética , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Mucosa Intestinal/metabolismo , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Lactobacillus plantarum/metabolismoRESUMO
Probiotics are gaining attention due to their functions of regulating the intestinal barrier and promoting human health. The production of bacteriocins is one of the important factors for probiotics to exert beneficial properties. This study aimed to screen bacteriocin-producing Lactiplantibacillus plantarum and evaluate the probiotic potential in vitro. It was found that L. plantarum Q7, L. plantarum F3-2 and L. plantarum YRL45 could produce bacteriocins and inhibit common intestinal pathogens. These three strains had probiotic potential with tolerance to the gastrointestinal environmental and colonization in the gut, and exhibited various degrees of anti-inflammatory activity and tight junction function in the intestinal barrier. Particularly, L. plantarum YRL45 could significantly (p < 0.05) reduce the increase in nitric oxide (NO), prostaglandin E2 (PGE2), necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) induced by lipopolysaccharide (LPS), thereby easing inflammatory response. L. plantarum F3-2 could remarkably (p < 0.05) up-regulate the expression levels of ZO-1, Occludin and Claudin-1 in intestinal epithelial injured cells, which was conducive to protecting the intestinal barrier. These findings provided fundamental information about the probiotic properties of bacteriocin-producing L. plantarum, which suggested that L. plantarum Q7, L. plantarum F3-2 and L. plantarum YRL45 had the potential to be used as novel probiotic strains.
RESUMO
Plantaricin Q7 is a bacteriocin produced by Lactobacillus plantarum Q7 with food preservation potential. Low yield is one of the bottlenecks of the wide application of plantaricin Q7. Nontargeted metabolomics was performed to reveal the mechanism of plantaricin Q7 biosynthesis. The results showed that the composition and abundance of intracellular metabolites varied significantly at key time points of plantaricin Q7 synthesis. Differential metabolic pathways were purine metabolism; pyrimidine metabolism; alanine, aspartate, and glutamate metabolism; amino acid biosynthesis; aminoacyl-tRNA biosynthesis; and ABC transporters. Differential metabolites were xanthine, deoxyadenosine, uracil, 5-methylcytosine, α-ketoglutarate, γ-aminobutyric acid, glutamate, glutamine, and tryptophan. Based on metabolomics information, the putative metabolic synthesis pathway of plantaricin Q7 was proposed. Glutamine, glutamate, and 5-methylcytosine could be critical metabolites and simulate plantaricin Q7 biosynthesis significantly (P < 0.05). Bacteriocin production was investigated by comparative metabolomics in this report, which could help to achieve higher plantaricin Q7 yield by metabolic regulation.
Assuntos
Bacteriocinas , Lactobacillus plantarum , Bacteriocinas/metabolismo , Vias Biossintéticas , Conservação de Alimentos , Lactobacillus plantarum/metabolismo , MetabolômicaRESUMO
The 2303 Wetlands of International Importance distribute unevenly in different continents. Europe owns the largest number of sites, while Africa has the largest area of sites. More than half of the sites are affected by three or four impact factors (55%). The most significant impact factors are pollution (54%), biological resources use (53%), natural system modification (53%), and agriculture and aquaculture (42%). The main affected objects are land area and environment of the wetlands, occurred in 75% and 69% of the sites, respectively. The types most affected by land area occupation are river wetlands and lake wetlands, the types with the greatest impact on environment are marine/coastal wetlands and river wetlands, the type with the greatest impact on biodiversity is river wetlands, the types most affected by water resources regulation are marsh wetlands and river wetlands, and the types most affected by climate change are lake wetlands and marine/coastal wetlands. About one-third of the wetland sites have been artificially reconstructed. However, it is found that the proportions of natural wetland sites not affected or affected by only one factor are generally higher than that of wetland sites both containing natural wetlands and human-made wetlands, while the proportions of wetland sites both containing natural wetlands and human-made wetlands affected by three or four factors are generally higher than that of natural wetland sites. Wetland sites in the UK and Ireland are least affected among all countries. Wetland management plans in different regions still have large space for improvement, especially in Africa and Asia. The protection and restoration of global wetlands can be carried out in five aspects, including management and policy, monitoring, restoration, knowledge, and funding.
Assuntos
Biodiversidade , Conservação dos Recursos Naturais , Monitoramento Ambiental , Áreas AlagadasRESUMO
Eosinophils are a type of thymic stromal cell that are present in the thymus of both humans and mice. They participate in regulating T-cell development under non-pathological conditions. However, studies are scarce regarding the role of eosinophils in the development of the thymus in chickens. Therefore, this study investigated the distribution of eosinophils in normal chicken thymi at different stages of development. Seven thymi were obtained from chickens at days 1, 21 and 35 of development. The distribution of eosinophils in the thymi was analyzed by histological and immunohistochemical techniques using Lendrum's chromotrope 2R method and an antibody against eosinophilic cationic protein (ECP), respectively. Eosinophils were constitutively located in the chick thymus. They were mainly distributed in the thymic corticomedullary junction and medulla, especially around vessels and Hassall's corpuscles, and only a few were in the trabeculae among thymic lobules and around vessels. There were none in the cortex. The number of thymic eosinophils decreased with increasing age (P<0.01). These results indicated that eosinophils comprise a type of thymic stromal cells in the chick, which may regulate thymic development, especially during the early stages of development.