Multi-omics analysis reveals the molecular regulatory network underlying the prevention of Lactiplantibacillus plantarum against LPS-induced salpingitis in laying hens

Background Salpingitis is one of the common diseases in laying hen production, which greatly decreases the economic outcome of laying hen farming. Lactiplantibacillus plantarum was effective in preventing local or systemic inflammation, however rare studies were reported on its prevention against salpingitis. This study aimed to investigate the preventive molecular regulatory network of microencapsulated Lactiplantibacillus plantarum(MLP) against salpingitis through multi-omics analysis, including microbiome, transcriptome and metabolome analyses.Results The results revealed that supplementation of MLP in diet significantly alleviated the inflammation and atrophy of uterus caused by lipopolysaccharide(LPS) in hens(P < 0.05). The concentrations of plasma IL-2 and IL-10 in hens of MLP-LPS group were higher than those in hens of LPS-stimulation group(CN-LPS group)(P < 0.05). The expression levels of TLR2, MYD88, NF-κB, COX2, and TNF-α were significantly decreased in the hens fed diet supplemented with MLP and suffered with LPS stimulation(MLP-LPS group) compared with those in the hens of CN-LPS group(P < 0.05). Differentially expressed genes(DEGs) induced by MLP were involved in inflammation, reproduction, and calcium ion transport. At the genus level, the MLP supplementation significantly increased the abundance of Phascolarctobacterium, whereas decreased the abundance of Candidatus_Saccharimonas in LPS challenged hens(P < 0.05). The metabolites altered by dietary supplementation with MLP were mainly involved in galactose, uronic acid, histidine, pyruvate and primary bile acid metabolism. Dietary supplementation with MLP inversely regulates LPSinduced differential metabolites such as Lyso PA(24:0/0:0)(P < 0.05).Conclusions In summary, dietary supplementation with microencapsulated Lactiplantibacillus plantarum prevented salpingitis by modulating the abundances of Candidatus_Saccharimonas, Phascolarctobacterium, Ruminococcus_torques_group and Eubacterium_hallii_group while downregulating the levels of plasma metabolites, p-tolyl sulfate, o-cresol and N-acetylhistamine and upregulating S-lactoylglutathione, simultaneously increasing the expressions of CPNE4, CNTN3 and ACAN genes in the uterus, and ultimately inhibiting oviducal inflammation.

Lactiplantibacillus plantarum FRT4 attenuates high-energy low-protein dietinduced fatty liver hemorrhage syndrome in laying hens through regulating gut-liver axis

Background Fatty liver hemorrhage syndrome(FLHS)becomes one of the most major factors resulting in the laying hen death for caged egg production.This study aimed to investigate the therapeutic effects of Lactiplantibacillus plan-tarum(Lp.plantarum)FRT4 on FLHS model in laying hen with a focus on liver lipid metabolism,and gut microbiota.Results The FLHS model of laying hens was established by feeding a high-energy low-protein(HELP)diet,and the treatment groups were fed a HELP diet supplemented with differential proportions of Lp.plantarum FRT4.The results indicated that Lp.plantarum FRT4 increased laying rate,and reduced the liver lipid accumulation by regulating lipid metabolism(lipid synthesis and transport)and improving the gut microbiota composition.Moreover,Lp.plan-tarum FRT4 regulated the liver glycerophospholipid metabolism.Meanwhile,“gut-liver”axis analysis showed that there was a correlation between gut microbiota and lipid metabolites.Conclusions The results indicated that Lp.plantarum FRT4 improved the laying performance and alleviated FLHS in HELP diet-induced laying hens through regulating“gut-liver”axis.Our findings reveal that glycerophospholipid metabolism could be the underlying mechanism for the anti-FLHS effect of Lp.plantarum FRT4 and for future use of Lp.plantarum FRT4 as an excellent additive for the prevention and mitigation of FLHS in laying hens.

Lactiplantibacillus plantarum AR113 alleviates microbiota dysbiosis of tongue coating and cerebral ischemia/reperfusion injury in rat

Stroke is one of the leading causes of death and disability worldwide.However,information on stroke-related tongue coating microbiome(TCM)is limited,and whether TCM modulation could benefit for stroke prevention and rehabilitation is unknown.Here,TCM from stroke patients(SP)was characterized using molecular techniques.The occurrence of stroke resulted in TCM dysbiosis with significantly reduced species richness and diversity.The abundance of Prevotella,Leptotrichia,Actinomyces,Alloprevotella,Haemophilus,and TM7_[G-1]were greatly reduced,but common infection Streptococcus and Pseudomonas were remarkably increased.Furthermore,an antioxidative probiotic Lactiplantibacillus plantarum AR113 was used for TCM intervention in stroke rats with cerebral ischemia/reperfusion(I/R).AR113 partly restored I/R induced change of TCM and gut microbiota with significantly improved neurological deficit,relieved histopathologic change,increased activities of antioxidant enzymes,and decreased contents of oxidative stress biomarkers.Moreover,the gene expression of antioxidant-related proteins and apoptosis-related factors heme oxygenase-1(HO-1),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),nuclear factor erythroid 2-related factor 2(Nrf2),NAD(P)H:quinone oxidoreductase-1(NQO-1),and Bcl-2 was significantly increased,but cytochrome C,cleaved caspase-3,and Bax were markedly decreased in the brain by AR113 treatment.The results suggested that AR113 could ameliorate cerebral I/R injury through antioxidation and anti-apoptosis pathways,and AR113 intervention of TCM may have the application potential for stroke prevention and control.

Modulatory effects of Lactiplantibacillus plantarum on chronic metabolic diseases

The increased global incidence of chronic metabolic diseases,a vital threat to human health and a burden on our healthcare systems,includes a series of clinical metabolic syndromes such as obesity,diabetes,hypertension,and dyslipidemia.One of the well-known probiotic microorganisms,Lactiplantibacillus plantarum plays an important role in promoting human health,including inhibiting the occurrence and development of a variety of chronic metabolic diseases.The present study provides an overview of the preventive and therapeutic effects of L.plantarum on diabetes,obesity,non-alcoholic fatty liver disease,kidney stone disease,and cardiovascular diseases in animal models and human clinical trials.Ingesting L.plantarum demonstrated its ability to reduce inflammatory and oxidative stress levels by regulating the production of cytokines and short-chain fatty acids(SCFAs),the activity of antioxidant enzymes,and the balance of intestinal microbial communities to alleviate the symptoms of chronic metabolic diseases.Furthermore,updated applications and technologies of L.plantarum in food and biopharmaceutical industries are also discussed.Understanding the characteristics and functions of L.plantarum will guide the development of related probiotic products and explore the modulatory benefit of L.plantarum supplementations on the prevention and treatment of multiple chronic metabolic diseases.

Efficacy and dose response of Lactiplantibacillus plantarum in diarrhea-predominant irritable bowel syndrome

BACKGROUND Probiotics have shown promise in alleviating symptoms of diarrhea-predominant irritable bowel syndrome(IBS-D);however,the certainty of evidence is low.Wellpowered randomized controlled dose-ranging trials are warranted on promising single-strain candidates.AIM To investigate the clinical efficacy of Lactiplantibacillus plantarum(L.plantarum)Lpla33(DSM34428)in adults with IBS-D.METHODS This is a randomized,double-blind,placebo-controlled,multi-center,and doseranging study.Three hundred and seven adults,18-70 years of age,with IBS-D,according to Rome IV criteria,were allocated(1:1:1)to receive placebo or L.plantarum Lpla33 at 1×10^(9)(1B)or 1×10^(10)(10B)colony-forming units/d over an 8-wk intervention period.The primary outcome was the change in IBS severity scoring system(IBS-SSS)total score after 8 wk,while secondary and exploratory outcomes included abdominal pain severity,IBS related quality of life,stool and microbial profile,and perceived stress.RESULTS IBS-SSS was significantly reduced,after 8 wk,in participants receiving L.plantarum 1B(-128.45±83.30;P<0.001)and L.plantarum 10B(-156.77±99.06;P<0.001),compared to placebo(-58.82±74.75).Further,a dose-ranging effect was observed,with a greater absolute reduction in the L.plantarum 10B group(P<0.05).A reduction in sub-scores related to abdominal pain,abdominal distension,bowel habits,and quality of life was observed in both L.plantarum groups compared to placebo(P<0.001).Further,62.5%and 88.4%of participants administered L.plantarum 1B and 10B,respectively,were classified as stool consistency responders based on a reduction in diarrheal stool form,as compared to 26.3%in the placebo group(P<0.001).In contrast,no significant shifts were observed in microbial diversity.CONCLUSION L.plantarum Lpla33(DSM34428)is well tolerated and improves IBS symptom severity with a dose-ranging effect and a corresponding normalization of bowel habits in adults with IBS-D.

Dietary Lactiplantibacillus plantarum KX041 attenuates colitis-associated tumorigenesis and modulates gut microbiota

Colorectal cancer(CRC)is one of the most common cancers and supplementation of probiotics may be a promising intervention method. The present study aimed to investigate the anti-CRC effects of Lactiplantibacillus plantarum KX041 on a CRC mouse model. The CRC mice were induced by 10 mg/kg azoxymethane and 2% dextran sulfate sodium. L. plantarum KX041 was orally administrated once daily(1 × 10^(9) CFU/mouse). Results showed that L. plantarum KX041 could significantly inhibit inflammation, tumor formation, and induce tumor cells apoptosis. Moreover, this probiotic could ameliorate the damage of intestinal barrier by recovering tight junction protein expression(like Occludin, Claudin-1, and ZO-1)and preventing goblet cell loss. Furthermore, the oxidative stress was alleviated by increasing the level of antioxidant mediators(like GSH and SOD)and reducing the level of oxidative mediators(like MDA and MPO). In addition, treatment with L. plantarum KX041 could directly regulate gut microbiota, thereby increasing the abundance of beneficial bacteria(like SCFAs-producing bacteria, Akkermansia)and decreasing the abundance of harmful bacteria(like pro-inflammatory bacteria, Parasutterella), which in turn raised SCFAs levels and lowered LPS levels. In conclusion, L. plantarum KX041 could effectively ameliorate CRC via reshaping intestinal microenvironment, alleviating inflammation, maintaining intestinal permeability, and attenuating oxidative stress.

The protective effect of oleic acid on Lactiplantibacillus plantarum during freeze-drying

Membrane characteristics are determined by the fatty acids composition,which affects survival rates after freeze-drying.However,it is unknown which composition provides the greatest effect.In this study,we found that the addition of Tween 80 and Tween 20 significantly increased survival rates of Lactiplantibacillus plantarum,which reached a maximum of 93.1%.Conversely,Tween 60 caused a significant decrease.We further found that the difference between the effects of adding Tween 80 and Tween 60 was the change in oleic acid contents.To verify the role of oleic acid,we used CRISPR-Cas9 to knock-out the key synthesis gene cla-er.The survival rates of L.plantarum AR113Δcla-er declined to 5.48%.The addition of oleic acid restored the rates to those of wild type strains.Moreover,the membrane integrity and fl uidity of knockout strains markedly decreased.This is the fi rst confi rmation that Tween 80 or Tween 20 increases the survival rate by increasing the content of oleic acid in the cell membrane.These fi ndings also indicated that oleic acid in cell membranes has a substantial protective effect on L.plantarum during freeze-drying.

Influences of Lactiplantibacillus plantarum and Saccharomyces cerevisiae fermentation on the nutritional components,flavor property and lipid-lowering effect of highland barley

Highland barley is a well-known cereal in Qinghai-Tibet Plateau area with high nutritional value,which has been reported to be a health-promoting grain for the obesity and the diabetes.Fermentation by certain microbiota can improve the flavor property and nutritional characteristics.In the present study,Lactiplantibacillus plantarum and Saccharomyces cerevisiae were singly or jointly applied to ferment highland barley,and the profile of volatile substances and lipid-lowering effects of the respective extracts were analyzed.Results indicated that either L.plantarum or S.cerevisiae or co-fermentation could consume the polysaccharides of highland barley to provide energy,and dramatically increase the contents of total protein and polyphenol.Gas chromatography-mass spectrometry(GC-MS)analysis revealed that the presence of S.cerevisiae was critical for production of the pleasant flavors,especially for the ethyl ester substances including hexadecanoic acid ethyl,hexanoic acid ethyl ester and so on.Meanwhile,we found that fermented highland barley extracts by L.plantarum exhibited stronger lipid-lowering effects in Caenorhabditis elegans than that by S.cerevisiae,while the co-fermentation not only emitted pleasant odors but also exerted high hypolipidemic function.In all,co-fermentation by L.plantarum and S.cerevisiae was proposed to be a promising processing to improve the flavor and functional properties of highland barley.

Multi-omics reveals the mechanisms underlying Lactiplantibacillus plantarum P8-mediated attenuation of oxidative stress in broilers challenged with dexamethasone

Oxidative stress is a common phenomenon in poultry production.Several molecules,including antioxidant genes,miRNAs,and gut microbiota metabolites,have been reported to participate in redox regulation.Lactiplantibacillus plantarum P8(P8)was shown to improve the antioxidant capacity of chickens,but the specific molecular mechanisms remain unclear.In this study,400 broilers were allocated to 4treatment groups:control diet(Con group),control diet+dexamethasone injection(DEX group),control diet containing 1×108CFU/g P8(P8 group),and control diet containing 1×108CFU/g P8+DEX injection(DEX_P8 group).Integrated analysis of the microbiome,metabolomics,and miRNAomics was conducted to investigate the roles of P8 in oxidative stress in broilers.Results demonstrated that P8supplementation significantly improved growth performance,jejunal morphology,and antioxidant function in DEX-treated broilers.Analysis of the gut microbiota revealed a higher abundance of Barnesiella(P=0.01)and Erysipelatoclostridium(P=0.05)in the DEX_P8 group than in the DEX group.Functional prediction indicated that certain pathways,including the phenylacetate degradation pathway,were enriched in the DEX_P8 group compared to the DEX group.Metabolites in the cecal contents were distinct between the groups.P8 supplementation increased the content of metabolites with antioxidant capacity,e.g.,urobilinogen(P<0.01),and decreased that of metabolites related to oxidative stress,e.g.,genistein(P<0.01).Functional prediction indicated that metabolites that differed between the DEX_P8and DEX groups were enriched in pathways including“tryptophan metabolism”and“primary bile acid biosynthesis”.The miRNAomics analysis further showed that,compared to the DEX group,several miRNAs in the jejunum,such as gga-miR-21-3p(P=0.03),were increased,whereas gga-miR-455-3p(P=0.02)was decreased in the DEX_P8 group.The PI3K-Akt,Ras,and Rap1 signaling pathways were enriched in the DEX_P8 group compared to the DEX group through KEGG analysis.Correlation analysis revealed potential interactions between growth performance,oxidation/antioxidation,jejunal morphology,gut microbiota,cecal content metabolites,and jejunal miRNAs.Overall,our results indicate that P8 supplementation may improve the growth performance,jejunal morphology and antioxidant capacity of DEX-treated broilers by regulating gut microbiota,its metabolites,and intestinal miRNAs.

A micro-sized vaccine based on recombinant Lactiplantibacillus plantarum fights against SARS-CoV-2 infection via intranasal immunization

COVID-19 has globally spread to burden the medical system.Even with a massive vaccination,a mucosal vaccine offering more comprehensive and convenient protection is imminent.Here,a micro-sized vaccine based on recombinant Lactiplantibacillus plantarum(rLP)displaying spike or receptor-binding domain(RBD)was characterized as microparticles,and its safety and protective effects against SARS-CoV-2 were evaluated.We found a 66.7%mortality reduction and 100%protection with rLP against SARS-CoV-2 in a mouse model.The histological analysis showed decreased hemorrhage symptoms and increased leukocyte infiltration in the lung.Especially,rLP:RBD significantly decreased pulmonary viral loads.For the first time,our study provides a L.plantarum-vectored vaccine to prevent COVID-19 progress and transmission via intranasal vaccination.

Lactiplantibacillus plantarum L47 and inulin alleviate enterotoxigenic Escherichia coli induced ileal inflammation in piglets by upregulating the levels of a-linolenic acid and 12,13-epoxyoctadecenoic acid

Alternatives to antibiotics for preventing bacteria-induced inflammation in early-weaned farm animals are sorely needed. Our previous study showed that Lactiplantibacillus plantarum L47 and inulin could alleviate dextran sulfate sodium(DSS)-induced colitis in mice. To explore the protective effects of L. plantarum L47 and inulin on the ileal inflammatory response in weaned piglets challenged with enterotoxigenic Escherichia coli(ETEC), 28 weaned piglets were assigned into four groups, namely, CON group—orally given 10 mL/d phosphate buffer saline(PBS), LI47 group—orally given a mixture of 10 m L/d L. plantarum L47 and inulin, ECON group—orally given 10 mL/d PBS and challenged by ETEC, and ELI47group—orally given 10 mL/d L. plantarum L47 and inulin mixture and challenged by ETEC. The results demonstrated that the combination of L. plantarum L47 and inulin reduced inflammatory responses and relieved the inflammatory damage caused by ETEC, including ileal morphological damage, reduced protein expression of ileal tight junction, decreased antioxidant capacity, and decreased antiinflammatory factors. Transcriptome analysis revealed that L. plantarum L47 and inulin up-regulated the gene expression of phospholipase A2 group IIA(PLA2G2A)(P < 0.05) as well as affected alphalinolenic acid(ALA) metabolism and linoleic acid metabolism. Moreover, L. plantarum L47 and inulin increased the levels of ALA(P < 0.05), lipoteichoic acid(LTA)(P < 0.05), and 12,13-epoxyoctadecenoic acid(12,13-EpOME)(P < 0.05) and the protein expression of Toll-like receptor 2(TLR2)(P = 0.05) in the ileal mucosa. In conclusion, L. plantarum L47 and inulin together alleviated ETEC-induced ileal inflammation in piglets by up-regulating the levels of ALA and 12,13-EpOME via the LTA/TLR2/PLA2G2A pathway.

Lactobacillus plantarum AR113 attenuates liver injury in D-galactose-induced aging mice via the inhibition of oxidative stress and endoplasmic reticulum stress

Probiotics could effectively eliminate excess reactive oxygen species(ROS)generated during aging or lipid metabolism disorders,but their mechanism is unclear.The major purpose of this study was to investigate the mechanism of Lactiplantibacillus plantarun AR113 alleviating oxidative stress injury in the D-galactose induced aging mice.The result showed that pretreatment with L.plantarun AR113 significantly relieving H_(2)O_(2)induced cytotoxicity in HepG2 cells by maintain cell membrane integrity and increasing antioxidant enzyme activities.In D-galactose induced aging mice,L.plantarun AR113 could significantly attenuate liver damage and inflammatory infiltration by promoting endogenous glutathione(GSH)synthesis and activating the Nrf2/Keap1 signaling pathway in mice,and increasing the expression of regulated phaseⅡdetoxification enzymes and antioxidant enzymes.Further analysis shown that gavage of L.plantarun AR113 could significantly reduce the expression of G protein-coupled receptor 78(GPR78)and C/EBP homologous protein(CHOP)proteins,and promote the restoration of endoplasmic reticulum(ER)homeostasis,thereby activating cell anti-apoptotic pathways.These results were also confirmed in H_(2)O_(2)-treated HepG2 experiments.It indicated that L.plantarun AR113 could inhibit D-galactose-induced liver injury through dual inhibition of ER stress and oxidative stress.L.plantarun AR113 have good application potential in anti-aging and alleviating metabolic disorders.

Oral administration of Lactiplantibacillus plantarum 22A-3 exerts anti-allergic activity against intestinal food allergy mouse models sensitized and challenged with ovalbumin

In recent years,researches on food components with anti-allergic effects have been gathering attention,because of the expectation for the establishment of a safe and effective treatment for food allergy.Previous studies have reported that Lactiplantibacillus plantarum 22A-3 (LP22A3) inhibited degranulation of mast cells and reduced IgE production.We have developed a gastrointestinal allergy system in which mice are sensitized by intraperitoneal and oral administration of OVA and then challenged by oral administration of high doses of OVA.As a result,an increase in the amount of IgE in the blood and a decrease in the temperature of the colon were confirmed,and it was clarified that food allergy was induced by oral administration of high dose of OVA as the challenge.Oral administration of LP22A3 ameliorated allergic responses significantly by reducing the amount of IgE in the blood and to recovered the decrease of rectal temperature.However,LP22A3 did not affect the intestinal barrier function.Administered LP22A3 significantly suppressed mRNA expression of OX40L and IL-4.These results suggested that LP22A3 suppressed Th2 differentiation and IL-4 production via downregulation of OX40L,and consequently suppressed IgE production.LP22A3 might provide a safe and effective treatment for allergic diseases due to ability modulating intestinal immune system.

Prevalence of Clostridium spp.,in Kashar cheese and efficiency of Lactiplantibacillus plantarum and Lactococcus lactis subsp.lactis mix as a biocontrol agents for Clostridium spp.

Clostridium sporogenes and Clostridium tepidium strains were detected in traditional Kashar cheese samples with late blowing characteristics.To control Clostridium spp.,in Kashar cheese,dairy originated Lactic Acid Bacteria(LAB)strains were tested under in vitro conditions and during Kashar production.Two strains,Lactiplantibacillus plantarum and Lactococcus lactis subsp.lactis demonstrated anticlostridial activity in vitro and the co-inoculum of these two strains(107 cfu g^(-1))were tested during the challenge test on Kashar cheese production in which a contamination ratio of 10^(4) cfu g^(-1) with spores of Cl.sporogenes were applied.Kashar samples were stored at 4℃ and 25℃ during 40 days of storage period and microbiological and physicochemical properties of Kashar samples were determined during this period.A decrement of nearly 1 log cfu g^(-1) in Cl.sporogenes numbers was observed in Kashar samples produced with co-inoculum of Lb.plantarum and L.lactis subsp.lactis stored at 4℃ but this was not the case for the Kashar samples stored at 25℃.This study revealed the potential of distinct LAB strains to control Cl.sporogenes spores in semi-hard cheese samples as biocontrol agents at 4℃ storage.

Characterization of genomic,physiological,and probiotic features Lactiplantibacillus plantarum DY46 strain isolated from traditional lactic acid fermented shalgam beverage

Lactiplantibacillus plantarum is a significant probiotic where it could be found in ubiquitous niches.In this study,a new Lb.plantarum strain DY46 was isolated from a traditional lactic-acid-fermented beverage called shalgam.The whole genome of the DY46 was sequenced and obtained sequences were assembled into a 3.32 Mb draft genome using PATRIC(3.6.8.).The DY46 genome consists of a single circular chromosome of 3,332,827 bp that is predicted to carry 3219 genes,including 61 tRNA genes,2 rRNA operons.The genome has a GC content of 44.3%includes 98 predicted pseudogenes,25 complete or partial transposases and 3 intact prophages.The genes encoding enzymes related in the intact EMP(Embden-Meyerhof-Parnas)and PK(phosphoketolase)pathways were predicted using BlastKOALA which is an indicator of having facultative heterofermentative pathways.DY46 genome also predicted to carry genes of Pln E,Pln F and Pln K showing the antimicrobial potential of this bacterium which can be linked to in vitro antagonism tests that DY46 can inhibit S.enterica sv.Typhimurium ATCC14028,K.pneumonie ATCC13883,and P.vulgaris ATCC8427.Moreover,it is determined that all resistome found in its genome were intrinsically originated and the strain was found to be tolerant to acid and bile concentrations by mimicking human gastrointestinal conditions.In conclusion,L.plantarum DY46 is a promising bacterium that appears to have certain probiotic properties,confirmed by“in vitro”and“in silico”analyses,and is a potential dietary supplement candidate that may provide functional benefits to the host.

Methionine addition improves the acid tolerance of Lactiplantibacillus plantarum by altering cellular metabolic flux,energy distribution,lipids composition

This paper reported a wine-derived lactic acid bacterium,Lactiplantibacillus plantarum XJ25,which exhibited higher cell viability under acid stress upon methionine supplementation.Cellular morphology and the composition of the cytomembrane phospholipids revealed a more solid membrane architecture presented in the acid-stressed cells treated with methionine supplementation.Transcriptional analysis showed L.plantarum XJ25 reduced methionine transport and homocysteine biosynthesis under acid stress.Subsequent overexpression assays proved that methio-nine supplementation could alleviate the cell toxicity from homocysteine accumulation under acid stress.Finally,L.plantarum XJ25 employed energy allocation strategy to response environmental changes by balancing the uptake methionine and adjusting saturated fatty acids(SFAs)in membrane.These data support a novel mechanism of acid resistance involving methionine utilization and cellular energy distribution in LAB and provide crucial theoretical clues for the mechanisms of acid resistance in other bacteria.

Proteomics analysis of the hypothalamus of high-fat diet fed mice after Lactiplantibacillus plantarum Y44 administration

Probiotics could regulate human health via changing gut microbiota.Our previous studies found that Lactiplantibacillus plantarum Y44 (L.plantarum Y44) was shown to reverse obesity-related symptoms of high-fat diet fed mice by influencing gut microbiota,which ameliorated lipid metabolism disorders and intestinal inflammation.In this study,the effects of L.plantarum Y44 on high-fat fed mice and the involved mechanisms were investigated through hypothalamus proteomics analysis,which revealed 22 downregulated and 21 upregulated proteins.Differentially expressed proteins between L.plantarum Y44 intervention group and high-fat fed group in hypothalamus of mice involved in metabolic pathways were concentrated in endocrine regulation of carbohydrate,amino acid and energy generation pathways.The hypothalamus proteomics results about the influences on most of differentially expressed proteins of L.plantarum Y44 administering high-fat fed mice has been verified.Therefore,L.plantarum Y44 would be a potential functional strain for the amelioration of high-fat diet associated health problems.

Carotenoid productivity in human intestinal bacteria Eubacterium limosum and Leuconostoc mesenteroides with functional analysis of their carotenoid biosynthesis genes

The human intestinal microbiota that comprise over 1,000 species thrive in dark and anaerobic environments.They are recognized for the production of diverse low-molecular-weight metabolites crucial to human health and diseases.Carotenoids,low-molecular-weight pigments known for their antioxidative activity,are delivered to humans through oral intake.However,it remains unclear whether human intestinal bacteria biosynthesize carotenoids as part of the in-situ microbiota.In this study,we investigated carotenoid synthesis genes in vari-ous human gut and probiotic bacteria.As a result,novel candidates,the crtM and crtN genes,were identified in the carbon monoxide-utilizing gut anaerobe Eubacterium limosum and the lactic acid bacterium Leuconostoc mesenteroides subsp.mesenteroides.These gene candidates were isolated,introduced into Escherichia coli,which synthesized a carotenoid substrate,and cultured aerobically.Structural analysis of the resulting carotenoids re-vealed that the crtM and crtN gene candidates of E.limosum and L.mesenteroides mediate the production of 4,4′-diaponeurosporene through 15-cis-4,4′-diapophytoene.Evaluation of the crtE-homologous genes in these bacteria indicated their non-functionality for C40-carotenoid production.E.limosum and L.mesenteroides,along with the known carotenogenic lactic acid bacterium Lactiplantibacillus plantarum,were observed to produce no carotenoids under strictly anaerobic conditions.The two lactic acid bacteria synthesized detectable levels of 4,4′-diaponeurosporene under semi-aerobic conditions.The findings highlight that the obligate anaerobe E.limo-sum retains aerobically functional C30-carotenoid biosynthesis genes,potentially with no immediate self-utility,suggesting an evolutionary direction in carotenoid biosynthesis.(229 words)