AIM： To investigate the capacity of potentially probiotic strains from six bacterial genera to induce cytokine production alone or in combinations in order to identify potential enhancing or synergistic effects in or...AIM： To investigate the capacity of potentially probiotic strains from six bacterial genera to induce cytokine production alone or in combinations in order to identify potential enhancing or synergistic effects in order to select probiotic bacteria for in vivo purposes.METHODS： Cytokine production in human peripheral blood mononuclear cells （PBMC） in response to stimulation with eleven different potentially probiotic bacterial strains from Streptococcus, Lactobacillus, Bifidobacterium, Lactococcus, L euconostoc a n d Propionibacterium genera was analysed. Production and mRNA expression of TNF-α, IL-12, IFN-y and IL-10 were determined by ELISA and Northern blotting, respectively.RESULTS： All tested bacteria induced TNF-α production. The best inducers of Thl type cytokines IL-12 and IFN-y were Streptococcus and Leuconostoc strains. All BiHdobacterium and Propionibacterium strains induced higher IL-IO production than other studied bacteria. Stimulation of PBMC with any bacterial combinations did not result in enhanced cytokine production suggesting that different bacteria whether gram-positive or gram- negative compete with each other during host cell interactions.CONCLUSION： The probiotic S. thermophilus and Leuconostoc strains are more potent inducers of Thl type cytokines IL-12 and IFN-γ than the probiotic Lactobacillus strains. Bacterial combinations did not result in enhanced cytokine production.展开更多
A strain with high production of exopolysaccharide(EPS) was isolated from dried milk cake(a traditional fermented food from Inner Mongolia). The strain was called N21 and later identified as Leuconostoc citreum( Leu. ...A strain with high production of exopolysaccharide(EPS) was isolated from dried milk cake(a traditional fermented food from Inner Mongolia). The strain was called N21 and later identified as Leuconostoc citreum( Leu. citreum). The strain was cultured in Man-Rogosa-Sharpe medium containing 50 g/L of sucrose for 48 h at 30 °C and the EPS purified, with a yield of 24.5 g/L. An average molecular weight of 6.07 × 10~6 g/mol was determined by high-performance size-exclusion chromatography. The structure of the purified EPS was investigated through gas chromatography, ~1H and ^(13)C nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. The results demonstrated a polysaccharide composed of D-glucopyranose units in a linear chain with consecutive α(1 → 6) linkages. No branching was found in the structure of the exopolysaccharide. The purified EPS showed high water solubility and emulsibility. Based on the thermogravimetric curve, the degradation temperature of the EPS was 308.47 °C, which suggested that the dextran in the study exhibited high thermal stability. The results indicated that Leu. citreum N21 could be widely used to produce linear EPS and that the EPS has potential applications in food science and processing as a food additive.展开更多
Exopolysaccharides can be produced by various bacteria and have important biological roles in bacterial survival depend on molecular weight,linkage,and conformation.In this study,Leuconostoc pseudomesenteroides G29 wa...Exopolysaccharides can be produced by various bacteria and have important biological roles in bacterial survival depend on molecular weight,linkage,and conformation.In this study,Leuconostoc pseudomesenteroides G29 was identified and found to produce two types of exopolysaccharides from sucrose including soluble and insoluble a-glucans.By regulation of pH above 5.5,soluble a-glucan production was increased to 38.4 g∙L^(-1) from 101.4 g∙L^(-1) sucrose with fewer accumulation of lactic acid and acetic acid.Simultaneously,the quantity of thick white precipitate,that is insoluble a-glucan,was also increased.Then,a-glucans were prepared by enzymatic reaction with crude glucansucrases from the supernatant of G29 fermentation broth and purified for structure analysis.Based on the integration analysis of FT-IR and NMR,it was observed that soluble a-glucan is a highly linear dextran with α-1,6 glycosidic bonds while the insoluble a-glucan has 93%of α-1,3 and 7%of α-1,6 glycosidic bond.The results extend our understanding of exopolysaccharides production by L.pseudomesenteroides,and this water insoluble α-1,3-glucan might have potential application as biomaterials and/or biochemicals.展开更多
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...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)展开更多
The spatiotemporal distributions of microbes in soil by different methods could affect the efficacy of the microbes to reduce the soil hydraulic conductivity.In this study,the specimens of bio-mediated sands were prep...The spatiotemporal distributions of microbes in soil by different methods could affect the efficacy of the microbes to reduce the soil hydraulic conductivity.In this study,the specimens of bio-mediated sands were prepared using three different methods,i.e.injecting,mixing,and pouring a given microbial so-lution onto compacted sand specimens.The hydraulic conductivity was measured by constant-head tests,while any soil microstructural changes due to addition of the microbes were observed by scan-ning electron microscope(SEM)and mercury intrusion porosimetry(MIP)tests.The amount of dextran concentration produced by microbes in each type of specimen was quantified by a refractometer.Results show that dextran production increased exponentially after 5-7 d of microbial settling with the supply of culture medium.The injection and mixing methods resulted in a similar amount and uniform dis-tribution of dextran in the specimens.The pouring method,however,produced a nonuniform distri-bution,with a higher concentration near the specimen surface.As the supply of culture medium discontinued,the dextran content near the surface produced by the pouring method decreased dramatically due to high competition for nutrients with foreign colonies.Average dextran concentration was negatively and correlated with hydraulic conductivity of bio-mediated soils exponentially,due to the clogging of large soil pores by dextran.The hydraulic conductivity of the injection and mixing cases did not change significantly when the supply of culture medium was absent.展开更多
To study the effects of mixed starter consisted of different fermentative type of lactic acid bacteria(LAB)on the fermentation of suansun,two lactic acid bacteria(Leuconostoc citreum NM-12 and Lactobacillus plantarum ...To study the effects of mixed starter consisted of different fermentative type of lactic acid bacteria(LAB)on the fermentation of suansun,two lactic acid bacteria(Leuconostoc citreum NM-12 and Lactobacillus plantarum L01)isolated from Chinese traditional fermented vegetable were used in the preparation of suansun.The fermentation was carried out at ambient temperature(around 25℃)for 96 h by inoculating different mixing ratios of LAB(inoculated fermentation)or using natural microbes(natural fermentation).The changes of pH,titratable acid(TA),microbe communities,free sugars,organic acids,nitrite and volatile compounds during fermentation were evaluated.Suansun treated with high Leuconostoc citreum ratio inoculation exhibited a quickly change in pH and TA,resulting from the rapid increase in the number of viable cells,at the early stage of fermentation and produced more mannitol(0.12–0.46 mg/mL)and acetic acid(0.93–3.56 mg/mL).However,Suansun treated with high Lactobacillus plantarum ratio inoculation had lower pH and higher TA at the later stage of fermentation and produced more lactic acid(5.32–7.68 mg/mL).No mannitol was detected in suansun when only Lactobacillus plantarum was inoculated in fermentation.No p-cresol was produced in the inoculated fermentation with mixed starter culture,in addition to the production of ethyl acetate and 2.3-butanedione,which had a positive effect on the flavor of suansun.In summary,this study demonstrated the application value of mixed starter consisted of different fermentative type of LAB.LAB types and mixing ratios greatly affected the types and concentration of metabolites in suansun fermentation.展开更多
Leuconostoc spp. （LSland LI1） isolated from sauerkraut and idli batter was selected for dextran production. To enhance the yield of dextran, effects of various parameters such as sucrose concentration, pH, temperatu...Leuconostoc spp. （LSland LI1） isolated from sauerkraut and idli batter was selected for dextran production. To enhance the yield of dextran, effects of various parameters such as sucrose concentration, pH, temperature, incubation and inoculum percentage were analyzed. The optimum sucrose concentration for the Leuconostoc spp. （LS1 and LI1） was found to be 15% and 25% respectively. Isolates produced maximum dextran after 20 h of incubation at 29℃ and the optimum pH was found between 8 and 8.5. The inoculum concentration of 7.5% was more favorable for the production of dextran by Leuconostoc spp. （LS1 and LI1）. The growth kinetic parameters were studied and compared for the strains LS1 and LI1. Mass production of dextran was carried out using a stirred tank batch reactor. FTIR analysis was done to determine the functional groups of dextran, sephadex is prepared by cross linking dextran using epichlorohydrin and the functional groups are determined by FTIR analysis.展开更多
基金The Research Council for Health of the Academy of Finland,the Sigrid Juselius Foundation and Valio Research Centre
文摘AIM： To investigate the capacity of potentially probiotic strains from six bacterial genera to induce cytokine production alone or in combinations in order to identify potential enhancing or synergistic effects in order to select probiotic bacteria for in vivo purposes.METHODS： Cytokine production in human peripheral blood mononuclear cells （PBMC） in response to stimulation with eleven different potentially probiotic bacterial strains from Streptococcus, Lactobacillus, Bifidobacterium, Lactococcus, L euconostoc a n d Propionibacterium genera was analysed. Production and mRNA expression of TNF-α, IL-12, IFN-y and IL-10 were determined by ELISA and Northern blotting, respectively.RESULTS： All tested bacteria induced TNF-α production. The best inducers of Thl type cytokines IL-12 and IFN-y were Streptococcus and Leuconostoc strains. All BiHdobacterium and Propionibacterium strains induced higher IL-IO production than other studied bacteria. Stimulation of PBMC with any bacterial combinations did not result in enhanced cytokine production suggesting that different bacteria whether gram-positive or gram- negative compete with each other during host cell interactions.CONCLUSION： The probiotic S. thermophilus and Leuconostoc strains are more potent inducers of Thl type cytokines IL-12 and IFN-γ than the probiotic Lactobacillus strains. Bacterial combinations did not result in enhanced cytokine production.
基金supported by the National Science and Technology Support Plan Project(No.2015BAD16B01)
文摘A strain with high production of exopolysaccharide(EPS) was isolated from dried milk cake(a traditional fermented food from Inner Mongolia). The strain was called N21 and later identified as Leuconostoc citreum( Leu. citreum). The strain was cultured in Man-Rogosa-Sharpe medium containing 50 g/L of sucrose for 48 h at 30 °C and the EPS purified, with a yield of 24.5 g/L. An average molecular weight of 6.07 × 10~6 g/mol was determined by high-performance size-exclusion chromatography. The structure of the purified EPS was investigated through gas chromatography, ~1H and ^(13)C nuclear magnetic resonance spectroscopy, and Fourier transform infrared spectroscopy. The results demonstrated a polysaccharide composed of D-glucopyranose units in a linear chain with consecutive α(1 → 6) linkages. No branching was found in the structure of the exopolysaccharide. The purified EPS showed high water solubility and emulsibility. Based on the thermogravimetric curve, the degradation temperature of the EPS was 308.47 °C, which suggested that the dextran in the study exhibited high thermal stability. The results indicated that Leu. citreum N21 could be widely used to produce linear EPS and that the EPS has potential applications in food science and processing as a food additive.
基金supported by Jiangsu Key Lab of Biomass-based Green Fuels and Chemicals Foundation(JSBEM2016010),Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture of China.
文摘Exopolysaccharides can be produced by various bacteria and have important biological roles in bacterial survival depend on molecular weight,linkage,and conformation.In this study,Leuconostoc pseudomesenteroides G29 was identified and found to produce two types of exopolysaccharides from sucrose including soluble and insoluble a-glucans.By regulation of pH above 5.5,soluble a-glucan production was increased to 38.4 g∙L^(-1) from 101.4 g∙L^(-1) sucrose with fewer accumulation of lactic acid and acetic acid.Simultaneously,the quantity of thick white precipitate,that is insoluble a-glucan,was also increased.Then,a-glucans were prepared by enzymatic reaction with crude glucansucrases from the supernatant of G29 fermentation broth and purified for structure analysis.Based on the integration analysis of FT-IR and NMR,it was observed that soluble a-glucan is a highly linear dextran with α-1,6 glycosidic bonds while the insoluble a-glucan has 93%of α-1,3 and 7%of α-1,6 glycosidic bond.The results extend our understanding of exopolysaccharides production by L.pseudomesenteroides,and this water insoluble α-1,3-glucan might have potential application as biomaterials and/or biochemicals.
文摘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)
基金The first author(V.Kamchoom)acknowledges the grant(Grant No.FRB66065/0258-RE-KRIS/FF66/53)from King Mongkut’s Insti-tute of Technology Ladkrabang(KMITL)and National Science,Research and Innovation Fund(NSRF)the grant under Climate Change and Climate Variability Research in Monsoon Asia(CMON3)from the National Research Council of Thailand(NRCT)(Grant No.N10A650844)the National Natural Science Foundation of China(NSFC).
文摘The spatiotemporal distributions of microbes in soil by different methods could affect the efficacy of the microbes to reduce the soil hydraulic conductivity.In this study,the specimens of bio-mediated sands were prepared using three different methods,i.e.injecting,mixing,and pouring a given microbial so-lution onto compacted sand specimens.The hydraulic conductivity was measured by constant-head tests,while any soil microstructural changes due to addition of the microbes were observed by scan-ning electron microscope(SEM)and mercury intrusion porosimetry(MIP)tests.The amount of dextran concentration produced by microbes in each type of specimen was quantified by a refractometer.Results show that dextran production increased exponentially after 5-7 d of microbial settling with the supply of culture medium.The injection and mixing methods resulted in a similar amount and uniform dis-tribution of dextran in the specimens.The pouring method,however,produced a nonuniform distri-bution,with a higher concentration near the specimen surface.As the supply of culture medium discontinued,the dextran content near the surface produced by the pouring method decreased dramatically due to high competition for nutrients with foreign colonies.Average dextran concentration was negatively and correlated with hydraulic conductivity of bio-mediated soils exponentially,due to the clogging of large soil pores by dextran.The hydraulic conductivity of the injection and mixing cases did not change significantly when the supply of culture medium was absent.
基金supported by the Guangxi colledge students’innovation and entrepreneurship training projects,China(S202110594095).
文摘To study the effects of mixed starter consisted of different fermentative type of lactic acid bacteria(LAB)on the fermentation of suansun,two lactic acid bacteria(Leuconostoc citreum NM-12 and Lactobacillus plantarum L01)isolated from Chinese traditional fermented vegetable were used in the preparation of suansun.The fermentation was carried out at ambient temperature(around 25℃)for 96 h by inoculating different mixing ratios of LAB(inoculated fermentation)or using natural microbes(natural fermentation).The changes of pH,titratable acid(TA),microbe communities,free sugars,organic acids,nitrite and volatile compounds during fermentation were evaluated.Suansun treated with high Leuconostoc citreum ratio inoculation exhibited a quickly change in pH and TA,resulting from the rapid increase in the number of viable cells,at the early stage of fermentation and produced more mannitol(0.12–0.46 mg/mL)and acetic acid(0.93–3.56 mg/mL).However,Suansun treated with high Lactobacillus plantarum ratio inoculation had lower pH and higher TA at the later stage of fermentation and produced more lactic acid(5.32–7.68 mg/mL).No mannitol was detected in suansun when only Lactobacillus plantarum was inoculated in fermentation.No p-cresol was produced in the inoculated fermentation with mixed starter culture,in addition to the production of ethyl acetate and 2.3-butanedione,which had a positive effect on the flavor of suansun.In summary,this study demonstrated the application value of mixed starter consisted of different fermentative type of LAB.LAB types and mixing ratios greatly affected the types and concentration of metabolites in suansun fermentation.
文摘Leuconostoc spp. （LSland LI1） isolated from sauerkraut and idli batter was selected for dextran production. To enhance the yield of dextran, effects of various parameters such as sucrose concentration, pH, temperature, incubation and inoculum percentage were analyzed. The optimum sucrose concentration for the Leuconostoc spp. （LS1 and LI1） was found to be 15% and 25% respectively. Isolates produced maximum dextran after 20 h of incubation at 29℃ and the optimum pH was found between 8 and 8.5. The inoculum concentration of 7.5% was more favorable for the production of dextran by Leuconostoc spp. （LS1 and LI1）. The growth kinetic parameters were studied and compared for the strains LS1 and LI1. Mass production of dextran was carried out using a stirred tank batch reactor. FTIR analysis was done to determine the functional groups of dextran, sephadex is prepared by cross linking dextran using epichlorohydrin and the functional groups are determined by FTIR analysis.
