The research was first to perform the utilization of novel and cheap pectin crude extract from Krung Kha Mao leaves （Cissampelos pareira L.） to immobilize cells of Lactobacillus casei subsp, rhamnosus TISTR 108 and ...The research was first to perform the utilization of novel and cheap pectin crude extract from Krung Kha Mao leaves （Cissampelos pareira L.） to immobilize cells of Lactobacillus casei subsp, rhamnosus TISTR 108 and Lactobacillus delbrueckii subsp, bulgaricus TISTR 1339 for lactic acid production, the optimum condition of immobilized cells was produced significantly at 5% of probability, that the highest lactic acid has been 38.50 and 33.66 g/L in steady state of whey medium for 96 and 108 h, respectively. Both strains were immobilized by 4% （w/w） of pectin crude extract from Krung Kha Mao （KKM pectin） leaves, 1.52 mm inner diameters of silicone tube and 5% （v/v） inoculum for immobilization. Efficiency of lactic acid production was compared by immobilized cells ofL. casei subsp, rhamnosus TISTR 108 when KKM pectin, commercial citrus pectin, commercial apple pectin and sodium alginate were used as supporting materials to produce lactic acid 38.50, 38.76, 30.43 and 34.56 g/L, respectively, the productivity of lactic acid has been 0.40, 0.40, 0.36 and 0.36 g/L h, respectively.展开更多
A Lactobacillus brevis CGMCC 1306 isolated from fresh milk without pasteurization was found to have higher glutamate decarboxylase （GAD） activity. An effective isolation and purification procedure of GAD from a cell...A Lactobacillus brevis CGMCC 1306 isolated from fresh milk without pasteurization was found to have higher glutamate decarboxylase （GAD） activity. An effective isolation and purification procedure of GAD from a cell-free extract of Lactobacillus brevis was developed, and the procedure included four steps： 30%-90% saturation （NH4）2SO4 fractional precipitation, Q sepharose FF anion-exchange chromatography, sephacryl S-200 gel filtration, and resource Q anion-exchange chromatography. Using this protocol, the purified GAD was demonstrated to possess electrophoretic homogeneity via SDS-PAGE. The purification fold and activity recovery of GAD were 43.78 and 16.95%, respectively. The molecular weight of the purified GAD was estimated to be approximately 62 kDa via SDS-PAGE. The optimum pH and temperature of the purified GAD were 4.4 and 37℃, respectively. The purified GAD had a half-life of 50rain at 45℃ and the Km value of the enzyme from Lineweaver-Burk plot was found to be 8.22.5＇-pyridoxal phosphate （PLP） had little effect on the regulation of its activity.展开更多
This research focused using novel substrate, longan Lactobacillus casei subsp, rhamnosus TISTR 108. The optimum juice as carbon source for continuous lactic acid production by medium for lactic acid production was pur...This research focused using novel substrate, longan Lactobacillus casei subsp, rhamnosus TISTR 108. The optimum juice as carbon source for continuous lactic acid production by medium for lactic acid production was pure longan juice with 120 g/L sugar concentration and among the different nitrogen sources were added to the longan juice （yeast extract, tryptic soy, urea, （NH4）2SO4 and NaNO3）, yeast extract had the most efficiency. Yeast extract （10 g/L） without adding minerals to longan juice could produced the maximum lactic acid concentration of 38.91 ± 0.190 g/L in 60 h and the yield of 0.460± 0.122 g/g with the productivity of 0.649± 0.002 g/Lh in 2 liters flask. Batch fermentation was conducted in 2 liters fermentor and 41.38± 0.030 g/L lactic acid was produced in 48 h with the yield of 0.398 ± 0.215 g/g and the productivity was 0.862 ± 0.001 g/L h. The continuous fermentation using 2 liters fermentor as a high productivity for lactic acid （1.091 ± 0.001 g/L h） was achieved at dilution rate （D） of 0.0685 h-1.展开更多
L-（＋）-lactic acid production was studied by immobilized Lactobacillus rhamnosus T1STR108 on crude pectin from Krung Kha Mao Leaves. Central composite design was employed to determine the maximum lactic acid product...L-（＋）-lactic acid production was studied by immobilized Lactobacillus rhamnosus T1STR108 on crude pectin from Krung Kha Mao Leaves. Central composite design was employed to determine the maximum lactic acid production of 42.88 g L-1 in predicted model with the factors at 4.11 g L1 of pectin, 6.05 mLLl inoculum and 1.09 mm of bead diameter. Statistical analyses demonstrated very high significance for the regression model, since the F-value computed 116.09 was much higher than the tabulated F-value 2.08 for the lactic acid production at 5% level for linear and quadratic polynomial regression models. The highest experimental lactic acid production was 43.57 g L^-1 at 96 h of fermentation, 1.58% higher than the predicted value.展开更多
Cow＇s milk constitutes a significant food for babies when mother＇s milk is insufficient. The purpose of this work is to offer a fermented milk, potentially hypoallergenic, using selected Lactobaeillus acidophilus an...Cow＇s milk constitutes a significant food for babies when mother＇s milk is insufficient. The purpose of this work is to offer a fermented milk, potentially hypoallergenic, using selected Lactobaeillus acidophilus and Bifidobacteria sp. for their proteolytic activities. The fermentation is evaluated by the rate of lactic acid production and by the bacterial enumeration at 45 ℃. The rate of acidification obtained by mixed cultures （La ＋ B. longum, 63.8 ± 3.5 °D） and （La ＋ B. breve, 43.4 ± 1.67 °D） compared with the control （milk） （18.6 ± 1.31 °D）. The result of the best hydrolysis was obtained by （La＋ B. longum） （154.88 ± 30.33 ug/mg） which corresponded to a better release of the a-NH2 functions （103.32 ± 12.81 umoles/mg） compared with control （307.2 ± 11.54 ug/mg and 10.25 ± 0.44 umoles/mg ）. The best synergy was obtained by （La ＋ B. bifidum） （13 × 10^6 cfu/mL and 60 × 10^6 cfu/mL） and reached 95 × 10^10 cfu/mL and 119 × 10^10 cfu/mg at the end. The electrophoresis of fermented milk revealed the presence of soluble proteins （a-Lactalbumin and β-Lactoglobulin）. The Enzyme Linked Immtmo Sorbent Assay showed the results of residual antigenic activities of β-Lg （1.4 ± 0.23 ug/mg）, a-La （0.012 ± 0.004 ug/mg） and bovin serum albumin （0.014 ± 0.005 ug/mg）by the associations （La ＋ B. longum）, （La ＋ B. bifidum） and （La ＋ B. bifidum） compared with the control （14.43 ± 5.91 ug/mg, 0.183 ± 0.062 gg/mg, 0.05 ± 0.008 ug/mg） respectively. Lactic fermentation reduced to a significant antigenic reactivity of principal whey milk proteins.展开更多
Mutagenesis of Lactobacillus casei subsp, rhamnosus Xl-12 after low power microwave irradiation was investigated. Under a microwave power of 400 W and irradiation length of 3 min, a mutated strain W4-3-9 with high-yie...Mutagenesis of Lactobacillus casei subsp, rhamnosus Xl-12 after low power microwave irradiation was investigated. Under a microwave power of 400 W and irradiation length of 3 min, a mutated strain W4-3-9 with high-yield L-lactic acid was obtained by screening. Compared with the starting strain X1-12, the L-lactic acid production of W4-3-9 was increased by 58.0% at a concentration of 115.8 g/L. The strain maintained the capability of producing a high L-lactic acid level after 10 generations. Cell surface morphology and DNA structures of parental and mutated strains were observed by atomic force microscopy （ AFM ）. Amplified fragment length polymorphism （AFLP） analysis suggested the difference in AFLP band pattern between the mutated and non-mutated strains. Sequencing and BLAST analysis revealed that the catalytic site of lactate dehydrogenase （DHL） was changed due to the microwave induced mutation.展开更多
Coriolus versicolor has been reported to have biological activities such as anticancer, antibacterial, antibiotics and immunopotentiation. The authors tried to ferment using lactobacilli associated with various biolog...Coriolus versicolor has been reported to have biological activities such as anticancer, antibacterial, antibiotics and immunopotentiation. The authors tried to ferment using lactobacilli associated with various biological functions and expected that fermented extract by lactobacilli will be superior to non-fermented extract in antioxidant activity. C. versicolor was extracted with 70% ethanol and then it was fermented by Lactobacillus acidophilus KCTC3164 and Lactobacillus plantarum KCTC3099, respectively. 1,l-dipheny｜-2-picrylhydrazyl （DPPH） scavenging activity, superoxide dismutase （SOD）-like activity and tetrazolium salt 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assays were performed. DPPH scavenging activity of fermented extract by L. acidophilus was higher than fermented extract by L. plantarum and antioxidant effect of fermented extract was 20%-30% higher than non-fermented extract. SOD-like activity was represented 18% in fermented extract by L. acidophilus, and was 27% in fermented extract by L. plantarum higher than non-fermented extract. As a result, this study suggests that it may need an efficient protocol for dominant bio-physiological activities such as extract volume or appropriate working volume of cultured cell or mixture condition. The fermented C. versicolor extract should provide with useful antioxidant agent in the human skin.展开更多
L-（＋）-lactic acid production was studied by immobilized Lactobacillus rhamnosus TISTR108 on crude pectin from Krung Kha Mao （Cissampelospareira L.） leaves. Central composite design was employed to determine the m...L-（＋）-lactic acid production was studied by immobilized Lactobacillus rhamnosus TISTR108 on crude pectin from Krung Kha Mao （Cissampelospareira L.） leaves. Central composite design was employed to determine the maximum lactic acid production of 45.40 g/L in predicted model （Y = 43.98 - 2.43X1 ＋ 1.02X2 ＋ 2.96X3 - 8.72X1^2 - 3.99X2^2 - 1.74X3^2） with the factors at 5.9 of cultural medium pH, 37.6 ℃ of process temperature and 202 rpm of liquid agitation. Statistical analyses demonstrated very high significance for the regression model fitted the data adequately and explained the lactic acid production, since the F-value computed 54.89 was much higher than the tabulated F-value 2.08 for the lactic acid production at 5% level for linear and quadratic polynomial regression models. The highest experimental lactic acid production was 46.91 g/L at 72 h of fermentation.展开更多
The ingestion of probiotic lactic acid bacteria has been evaluated and noted that it has an effect on the balance of desirable microbiota in the gastrointestinal tract. Lactobacillus gasseri demonstrates good survival...The ingestion of probiotic lactic acid bacteria has been evaluated and noted that it has an effect on the balance of desirable microbiota in the gastrointestinal tract. Lactobacillus gasseri demonstrates good survival in the gastrointestinal tract, and it has been associated with a variety of probiotic activities and roles, including the reduction of fecal mutagenic enzymes, the production of bacteriocins and the stimulation of macrophages immunomodulation. The aim of the study was to evaluate the effects of a pool of L. gasseri strains isolated from the feces of breastfed infants added in the human milk of healthy women. The milk was both pasteurized and unpasteurized, to verify the cell cytotoxicity of macrophages and to quantify the production of immunologic mediators such as IL-4, IL-6, IFN-γ, TNF-α, NO and oxygen intermediary compounds （H2O2）. The administration of raw human milk and pasteurized human milk to infants is a regular, encouraged practice in units of intensive therapy （UITs） and our present investigation verified the beneficial effect of addition of a pool of L. gasseri to pasteurized human milk （PHML）. Our results show that probiotic supplementation helped to maintain cell viability, reduced IL-6 and IFN-γ production and stimulated TNF-α, NO, H2O2, IL-4 production. Nevertheless, the results indicate that the addition of lactobacillus to human milk was not a determinant in the production of TNF-α. L. gasseri added to breast milk did not present a cytotoxic risk, and the addition ofL. gasseri to pasteurized milk of human milk bank would benefit newborns that depend on milk banks for the colonization of more desirable microbiota.展开更多
Naturally fermented pickles harbour many lactic acid bacteria （LAB）. Forty-three LAB strains with conjugated linoleic acid （CLA）-producing ability were isolated from three naturally fermented pickle brines. Of the...Naturally fermented pickles harbour many lactic acid bacteria （LAB）. Forty-three LAB strains with conjugated linoleic acid （CLA）-producing ability were isolated from three naturally fermented pickle brines. Of these isolates, Ip15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability （26.1% conversion） at 48 h in de Man Rogosa Sharpe （MRS） broth in the presence of 100 IJg/ml of linoleic acid （LA）. Compared to other strains, L. plantarum strain Ip15 showed the highest tolerance upon increased levels of LA in the medium, i.e., up to 600 μg/ml. This strain converted about 25% of LA into CLA isomers [predominantly cis-9, trans-11 CLA （9-CLA） and trans-lO, cis-12 CLA （10-CLA）], of which 75% was 9-CLA. Interestingly, though the conversion rate of LA into CLA by Ip15 remained stable between 100 to 600μg/ml LA levels in the medium, it dropped sharply at 1000 μg/ml. Taken together, the Ip15 strain displayed relatively high LA tolerance with higher conversion rate, which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.展开更多
文摘The research was first to perform the utilization of novel and cheap pectin crude extract from Krung Kha Mao leaves （Cissampelos pareira L.） to immobilize cells of Lactobacillus casei subsp, rhamnosus TISTR 108 and Lactobacillus delbrueckii subsp, bulgaricus TISTR 1339 for lactic acid production, the optimum condition of immobilized cells was produced significantly at 5% of probability, that the highest lactic acid has been 38.50 and 33.66 g/L in steady state of whey medium for 96 and 108 h, respectively. Both strains were immobilized by 4% （w/w） of pectin crude extract from Krung Kha Mao （KKM pectin） leaves, 1.52 mm inner diameters of silicone tube and 5% （v/v） inoculum for immobilization. Efficiency of lactic acid production was compared by immobilized cells ofL. casei subsp, rhamnosus TISTR 108 when KKM pectin, commercial citrus pectin, commercial apple pectin and sodium alginate were used as supporting materials to produce lactic acid 38.50, 38.76, 30.43 and 34.56 g/L, respectively, the productivity of lactic acid has been 0.40, 0.40, 0.36 and 0.36 g/L h, respectively.
基金Supported by the National Natural Science Foundation of China (No.30570411)the Research Plan of Zhejiang Province, China.
文摘A Lactobacillus brevis CGMCC 1306 isolated from fresh milk without pasteurization was found to have higher glutamate decarboxylase （GAD） activity. An effective isolation and purification procedure of GAD from a cell-free extract of Lactobacillus brevis was developed, and the procedure included four steps： 30%-90% saturation （NH4）2SO4 fractional precipitation, Q sepharose FF anion-exchange chromatography, sephacryl S-200 gel filtration, and resource Q anion-exchange chromatography. Using this protocol, the purified GAD was demonstrated to possess electrophoretic homogeneity via SDS-PAGE. The purification fold and activity recovery of GAD were 43.78 and 16.95%, respectively. The molecular weight of the purified GAD was estimated to be approximately 62 kDa via SDS-PAGE. The optimum pH and temperature of the purified GAD were 4.4 and 37℃, respectively. The purified GAD had a half-life of 50rain at 45℃ and the Km value of the enzyme from Lineweaver-Burk plot was found to be 8.22.5＇-pyridoxal phosphate （PLP） had little effect on the regulation of its activity.
文摘This research focused using novel substrate, longan Lactobacillus casei subsp, rhamnosus TISTR 108. The optimum juice as carbon source for continuous lactic acid production by medium for lactic acid production was pure longan juice with 120 g/L sugar concentration and among the different nitrogen sources were added to the longan juice （yeast extract, tryptic soy, urea, （NH4）2SO4 and NaNO3）, yeast extract had the most efficiency. Yeast extract （10 g/L） without adding minerals to longan juice could produced the maximum lactic acid concentration of 38.91 ± 0.190 g/L in 60 h and the yield of 0.460± 0.122 g/g with the productivity of 0.649± 0.002 g/Lh in 2 liters flask. Batch fermentation was conducted in 2 liters fermentor and 41.38± 0.030 g/L lactic acid was produced in 48 h with the yield of 0.398 ± 0.215 g/g and the productivity was 0.862 ± 0.001 g/L h. The continuous fermentation using 2 liters fermentor as a high productivity for lactic acid （1.091 ± 0.001 g/L h） was achieved at dilution rate （D） of 0.0685 h-1.
文摘L-（＋）-lactic acid production was studied by immobilized Lactobacillus rhamnosus T1STR108 on crude pectin from Krung Kha Mao Leaves. Central composite design was employed to determine the maximum lactic acid production of 42.88 g L-1 in predicted model with the factors at 4.11 g L1 of pectin, 6.05 mLLl inoculum and 1.09 mm of bead diameter. Statistical analyses demonstrated very high significance for the regression model, since the F-value computed 116.09 was much higher than the tabulated F-value 2.08 for the lactic acid production at 5% level for linear and quadratic polynomial regression models. The highest experimental lactic acid production was 43.57 g L^-1 at 96 h of fermentation, 1.58% higher than the predicted value.
文摘Cow＇s milk constitutes a significant food for babies when mother＇s milk is insufficient. The purpose of this work is to offer a fermented milk, potentially hypoallergenic, using selected Lactobaeillus acidophilus and Bifidobacteria sp. for their proteolytic activities. The fermentation is evaluated by the rate of lactic acid production and by the bacterial enumeration at 45 ℃. The rate of acidification obtained by mixed cultures （La ＋ B. longum, 63.8 ± 3.5 °D） and （La ＋ B. breve, 43.4 ± 1.67 °D） compared with the control （milk） （18.6 ± 1.31 °D）. The result of the best hydrolysis was obtained by （La＋ B. longum） （154.88 ± 30.33 ug/mg） which corresponded to a better release of the a-NH2 functions （103.32 ± 12.81 umoles/mg） compared with control （307.2 ± 11.54 ug/mg and 10.25 ± 0.44 umoles/mg ）. The best synergy was obtained by （La ＋ B. bifidum） （13 × 10^6 cfu/mL and 60 × 10^6 cfu/mL） and reached 95 × 10^10 cfu/mL and 119 × 10^10 cfu/mg at the end. The electrophoresis of fermented milk revealed the presence of soluble proteins （a-Lactalbumin and β-Lactoglobulin）. The Enzyme Linked Immtmo Sorbent Assay showed the results of residual antigenic activities of β-Lg （1.4 ± 0.23 ug/mg）, a-La （0.012 ± 0.004 ug/mg） and bovin serum albumin （0.014 ± 0.005 ug/mg）by the associations （La ＋ B. longum）, （La ＋ B. bifidum） and （La ＋ B. bifidum） compared with the control （14.43 ± 5.91 ug/mg, 0.183 ± 0.062 gg/mg, 0.05 ± 0.008 ug/mg） respectively. Lactic fermentation reduced to a significant antigenic reactivity of principal whey milk proteins.
基金Sponsored by the National Natural Science Foundation of China(Grant No.50778053)
文摘Mutagenesis of Lactobacillus casei subsp, rhamnosus Xl-12 after low power microwave irradiation was investigated. Under a microwave power of 400 W and irradiation length of 3 min, a mutated strain W4-3-9 with high-yield L-lactic acid was obtained by screening. Compared with the starting strain X1-12, the L-lactic acid production of W4-3-9 was increased by 58.0% at a concentration of 115.8 g/L. The strain maintained the capability of producing a high L-lactic acid level after 10 generations. Cell surface morphology and DNA structures of parental and mutated strains were observed by atomic force microscopy （ AFM ）. Amplified fragment length polymorphism （AFLP） analysis suggested the difference in AFLP band pattern between the mutated and non-mutated strains. Sequencing and BLAST analysis revealed that the catalytic site of lactate dehydrogenase （DHL） was changed due to the microwave induced mutation.
文摘Coriolus versicolor has been reported to have biological activities such as anticancer, antibacterial, antibiotics and immunopotentiation. The authors tried to ferment using lactobacilli associated with various biological functions and expected that fermented extract by lactobacilli will be superior to non-fermented extract in antioxidant activity. C. versicolor was extracted with 70% ethanol and then it was fermented by Lactobacillus acidophilus KCTC3164 and Lactobacillus plantarum KCTC3099, respectively. 1,l-dipheny｜-2-picrylhydrazyl （DPPH） scavenging activity, superoxide dismutase （SOD）-like activity and tetrazolium salt 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assays were performed. DPPH scavenging activity of fermented extract by L. acidophilus was higher than fermented extract by L. plantarum and antioxidant effect of fermented extract was 20%-30% higher than non-fermented extract. SOD-like activity was represented 18% in fermented extract by L. acidophilus, and was 27% in fermented extract by L. plantarum higher than non-fermented extract. As a result, this study suggests that it may need an efficient protocol for dominant bio-physiological activities such as extract volume or appropriate working volume of cultured cell or mixture condition. The fermented C. versicolor extract should provide with useful antioxidant agent in the human skin.
文摘L-（＋）-lactic acid production was studied by immobilized Lactobacillus rhamnosus TISTR108 on crude pectin from Krung Kha Mao （Cissampelospareira L.） leaves. Central composite design was employed to determine the maximum lactic acid production of 45.40 g/L in predicted model （Y = 43.98 - 2.43X1 ＋ 1.02X2 ＋ 2.96X3 - 8.72X1^2 - 3.99X2^2 - 1.74X3^2） with the factors at 5.9 of cultural medium pH, 37.6 ℃ of process temperature and 202 rpm of liquid agitation. Statistical analyses demonstrated very high significance for the regression model fitted the data adequately and explained the lactic acid production, since the F-value computed 54.89 was much higher than the tabulated F-value 2.08 for the lactic acid production at 5% level for linear and quadratic polynomial regression models. The highest experimental lactic acid production was 46.91 g/L at 72 h of fermentation.
文摘The ingestion of probiotic lactic acid bacteria has been evaluated and noted that it has an effect on the balance of desirable microbiota in the gastrointestinal tract. Lactobacillus gasseri demonstrates good survival in the gastrointestinal tract, and it has been associated with a variety of probiotic activities and roles, including the reduction of fecal mutagenic enzymes, the production of bacteriocins and the stimulation of macrophages immunomodulation. The aim of the study was to evaluate the effects of a pool of L. gasseri strains isolated from the feces of breastfed infants added in the human milk of healthy women. The milk was both pasteurized and unpasteurized, to verify the cell cytotoxicity of macrophages and to quantify the production of immunologic mediators such as IL-4, IL-6, IFN-γ, TNF-α, NO and oxygen intermediary compounds （H2O2）. The administration of raw human milk and pasteurized human milk to infants is a regular, encouraged practice in units of intensive therapy （UITs） and our present investigation verified the beneficial effect of addition of a pool of L. gasseri to pasteurized human milk （PHML）. Our results show that probiotic supplementation helped to maintain cell viability, reduced IL-6 and IFN-γ production and stimulated TNF-α, NO, H2O2, IL-4 production. Nevertheless, the results indicate that the addition of lactobacillus to human milk was not a determinant in the production of TNF-α. L. gasseri added to breast milk did not present a cytotoxic risk, and the addition ofL. gasseri to pasteurized milk of human milk bank would benefit newborns that depend on milk banks for the colonization of more desirable microbiota.
基金Project (No. 2007AA100402) supported by the National High-Tech R&D Program (863) of China
文摘Naturally fermented pickles harbour many lactic acid bacteria （LAB）. Forty-three LAB strains with conjugated linoleic acid （CLA）-producing ability were isolated from three naturally fermented pickle brines. Of these isolates, Ip15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability （26.1% conversion） at 48 h in de Man Rogosa Sharpe （MRS） broth in the presence of 100 IJg/ml of linoleic acid （LA）. Compared to other strains, L. plantarum strain Ip15 showed the highest tolerance upon increased levels of LA in the medium, i.e., up to 600 μg/ml. This strain converted about 25% of LA into CLA isomers [predominantly cis-9, trans-11 CLA （9-CLA） and trans-lO, cis-12 CLA （10-CLA）], of which 75% was 9-CLA. Interestingly, though the conversion rate of LA into CLA by Ip15 remained stable between 100 to 600μg/ml LA levels in the medium, it dropped sharply at 1000 μg/ml. Taken together, the Ip15 strain displayed relatively high LA tolerance with higher conversion rate, which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.
