Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was ...Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was studied in suspension cultures, and it was found that oligosaccharide triggered cell apoptosis. Under transmission electron microscope the following morphological changes were observed: cell shrinkage, condensation of cytoplasm, nuclear fragmentation, and the increase of high electron density bodies in vacuole in great quantity. In oligosaccharide_treated cells, agarose gel electrophoresis revealed that DNA was digested into oligonucleosomal fragments that were times of 200 bp appearing as DNA ladders. Control cells were in normal physiological state, they were intact, abundant in organelle and with integral nucleus DNA, and the rate of taxol biosynthesis in these cells was very low. After the oligosaccharide to the culture system, the defense system of cells was elicited and the secondary metabolism was strengthened, i.e. phenolics were accumulated in the medium, the activity of polyphenol oxidase (PPO) was increased quickly and secondary wall of cells was thickened. The activity of L _phenylalanine ammonia lyase (PAL), the critical enzyme of the phenylpropanoid pathway, was increased promptly 1 h after elicitation. The rate of taxol production was improved sharply and the maximal taxol concentration at 72 h was six times that of control. Appearance of cell apoptosis was accompanied with the highest concentration of taxol in suspension cultures.展开更多
Yeast strain Y68 producing high level of pullulan was isolated from the phyton collected in Toulouse, France. This strain was identified to be Rhodotorula bacarum by BIOLOG analysis. This is the first report that pull...Yeast strain Y68 producing high level of pullulan was isolated from the phyton collected in Toulouse, France. This strain was identified to be Rhodotorula bacarum by BIOLOG analysis. This is the first report that pullulan was produced by Rhodotorula bacarum. The optimal medium (g L -1) for pullulan production by this strain was 80 glucose, 20 soybean cake hydrolysate, 5 K 2HPO 4, 1 NaCl, 0.2 MgSO 4·7H 2O, 0.6 (NH 4) 2SO 4, pH 7.0. Under this condition, 54 g L -1 pullulan was produced within 60 h at 30 ℃. Pullulan is a better starting material for producing marine prodrugs.展开更多
AIM： To isolate and identify the soybean conglycinin peptides that selectively stimulates the growth of bifidobacteria in vitro, and to investigate the effect of soybean conglycinin peptides on intestinal ecosystem i...AIM： To isolate and identify the soybean conglycinin peptides that selectively stimulates the growth of bifidobacteria in vitro, and to investigate the effect of soybean conglycinin peptides on intestinal ecosystem in vivo. METHODS： Soybean conglycinin was purified from soybean seeds by gel filtration （Sepharose-CL-6B）. These proteins were submitted to hydrolysis by pepsin. Several growth-stimulating peptides for bifidobacteria were isolated chromatographically from pepsin hydrolysis of soybean conglycinin and identified by means of matrixassisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. Parallel to in vitro study, in vivo experiments with soybean conglycinin peptides were performed in mice. Ninety male KM mice were randomly assigned into five groups of 16 mice each, and each group was administered for 21d intragastrically with physiological saline （control）, conglycinin, pepsin-treated conglycinin （PTC）, the most active fraction which isolated from pepsin-treated conglycinin （P2-PTC） and HCl-full hydrolysis of conglycinin （HCl-FHC）, respectively. Intestinal microflora were evaluated by standard microbiologic methods and biochemical assays of cecal content samples after treatment. RESULTS： The results showed that the peptides which were isolated from soybean conglycinin could stimulate the growth of bifidobacteria in vitro, and the molecular mass of purified peptides with MALDI-TOF-MS ranged from 693.32 to 1829.55. Compared with control group, in vivo experiments showed that P2-PTC group decreased cecal pH （7.08±0.08 vs7.21±0.09, P〈0.05） and enterococcicounts （5.38±0.26 log10CFU/g vs 5.78±0.19 log10CFU/g, P〈0.05）, significantly increased sIgA level （172.08±35.40 ng/g vs 118.27±33.93 ng/g, P〈0.01） and β-galactosidase activity （1.28±0.23 U/g vs 1.82±0.58 U/g, P〈0.05） CONCLUSION： The results have shown that conglycinin is good source for enzyme-mediated production of peptides which stimulate the growth of bifidobacteria. These peptides are inactive within the sequence of the parent protein but can be released during enzymatic hydrolysis, and in vivo experiments demonstrate that conglycinin peptides may be beneficial for improving gastrointestinal health.展开更多
Soy protein isolate/carboxymethyl chitosan （SPI/CMCH） blended films incorporated with glycerol were prepared using solution casting to investigate the effects of the SPI and CMCH ratios （100：0, 88：12, 67：33, 50...Soy protein isolate/carboxymethyl chitosan （SPI/CMCH） blended films incorporated with glycerol were prepared using solution casting to investigate the effects of the SPI and CMCH ratios （100：0, 88：12, 67：33, 50：50, 33：67, 12：88, 0：100） on the water sorption isotherm. The moisture sorption isotherm of the SPI/CMCH blended films was determined using various relative humidity＇s （16%, 35%, 55% and 76% RH） at 25 ± 1℃. The isotherms showed that the equilibrium moisture content （EMC） of the films increased with increasing CMCH content and the EMC value sharply increased above aw = 0.55. Understanding of sorption isotherms is important for prediction of moisture sorption properties of films via moisture sorption empirical models. The Guggenheim-Oswin, Brunauer-Emmett-Teller （BET）, and Anderson-de Boer （GAB） sorption model predictions were tested against the experimental data. The root mean square （RMS） values from the Oswin, BET, and GAB models respectively ranged from 698.54 to 1,557.54, 38.85 to 58.30, and 52.52 to 95.95. Therefore, the BET model was found to be the best-fit model for SPI/CMCH blended films at 25 ± 1 ℃.展开更多
Objective To explore the pharmacological mechanism of active saponin compounds of Tuchao Baibiandouren(Lablab Semen Album fried with earth,TCBBDR)in treating type 2 diabetes(T2DM)using UHPLC-Q-Exactive Orbitrap MS and...Objective To explore the pharmacological mechanism of active saponin compounds of Tuchao Baibiandouren(Lablab Semen Album fried with earth,TCBBDR)in treating type 2 diabetes(T2DM)using UHPLC-Q-Exactive Orbitrap MS and network pharmacology.Methods UHPLC-Q-Exactive Orbitrap MS was used for a qualitative analysis of saponin compounds in TCBBDR.PharmMapper and CTD were used to screen drug active compounds and disease targets,and an active compound-target network was constructed via Cytoscape 3.8.0.Molecular docking was applied with the drug active compounds and key targets using AutoDock Vina 1.1.2,and a trajectory for the molecular dynamics simulation was completed by GROMACS 2019-3.Results Sixteen saponin compounds were identified from TCBBDR,along with 292 saponin compoud targets and 792 T2DM targets.Through Venn analysis of target saponin constituents and T2DM related targets,a total of 91 intersection targets were screened out in the treatment of T2DM with saponin.The mean values of degree,betweenness centrality and closeness centrality were taken as the thresholds to screen out 22 key genes,among which 4 key proteins namely MAPK1,IGF1 EGFR,PIK3R1 were selected in the top 10 key genes.On this basis,the saponin active constituent-target-signaling pathway network was established.The Gene Ontology(GO)enrichment analysis showed that the related biological modules included activity of steroid hormone receptor,steroid binding,and insulin receptor binding,etc.;the related signaling pathways were EGFR,PI3K-Akt and MAPK,etc.;regulating signaling pathways like MAPK could induce the proliferation,inhibition and apoptosis of pancreaticβcells,increase the quantity of pancreaticβcells,improve the functions of pancreaticβcells and stimulate the insulin secretion.Docking experiment analysis showed that all selected saponin compounds could enter the active sites of targets and form 3–14 hydrogen bonds with residues of the active sites.Moreover,van der Waals forces were present between chemical compounds and active sites.By combining the docking binding energy,we determined that the chemical compounds showed strong binding energy to the targets.Conclusion TCBBDR exerts therapeutic effects on diabetes through multi-compound and multi-target collaboration.Specifically,saponin components mediate pathways including inflammatory reaction and signal transduction to treat T2DM by regulating several key proteins that interact with EGFR and a series of signaling pathways related to disease development.展开更多
Soybean stachyose (SBS) and phytic acid (PA) are anti-nutritional factors (ANF) which have deleterious effects on the growth and digestibility in fish. The present research studied the effects of dietary SBS and PA on...Soybean stachyose (SBS) and phytic acid (PA) are anti-nutritional factors (ANF) which have deleterious effects on the growth and digestibility in fish. The present research studied the effects of dietary SBS and PA on the expression of three serine protease genes in the liver of Japanese flounder (Paralichthys olivaceus). These genes are trypsinogen 1 (poTRY), elastase 1 (poEL) and chymotrypsinogen 1 (poCTRY). Eight artificial diets with graded levels of supplemented ANFs were formulated to 4 levels of SBS (0.00, 0.40, 0.80 and 1.50%), 4 levels of PA (0.00, 0.20, 0.40 and 0.80), respectively.Japanese flounder (initial weight 2.45 g ± 0.01 g) were fed with these diets for 10 weeks with three replications per treatment. At the end of 10 weeks, supplementation of 0.40% of dietary SBS or PA significantly increased the gene expression of poTRY and poCTRY (P<0.05). The same level of dietary SBS significantly decreased the gene expression of poEL. In comparison with the control group (ANF-free),dietary PA (0.2% and 0.8%) significantly decreased the gene expression of poTRY, poCTRY and poEL (P<0.05). However, excessive supplement of dietary SBS (1.5%) has no significant effects on these gene expressions (P>0.05). These results suggested that dietary SBS and dietary PA could directly affect the serine protease genes at the transcriptional level in Japanese flounder, and these genes'expression was more sensitive to dietary PA than to SBS under the current experimental conditions.展开更多
Using Undaria pinnatifida juice and soya bean as main material, glucono delta lactone （GDL） as coagulant, we produced light green and very nourishing Undaria pinnatifida bean curd. Using the single factor experiment...Using Undaria pinnatifida juice and soya bean as main material, glucono delta lactone （GDL） as coagulant, we produced light green and very nourishing Undaria pinnatifida bean curd. Using the single factor experiment to determine the quantity and concentration of Undaria pinnatifida juice milk, process conditions of coagulation is determined by three factors three levels experiment, the best formulation of Undaria pinnatifida tofu is determined by test of four factors and three levels, process conditions are： the addition of Undaria pinnatifida is 20%, soya-bean milk concentration is1：4.5, the amount of GDL is0.30%, solidification time is 15min, solidification temperature is 95 ℃.展开更多
文摘Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was studied in suspension cultures, and it was found that oligosaccharide triggered cell apoptosis. Under transmission electron microscope the following morphological changes were observed: cell shrinkage, condensation of cytoplasm, nuclear fragmentation, and the increase of high electron density bodies in vacuole in great quantity. In oligosaccharide_treated cells, agarose gel electrophoresis revealed that DNA was digested into oligonucleosomal fragments that were times of 200 bp appearing as DNA ladders. Control cells were in normal physiological state, they were intact, abundant in organelle and with integral nucleus DNA, and the rate of taxol biosynthesis in these cells was very low. After the oligosaccharide to the culture system, the defense system of cells was elicited and the secondary metabolism was strengthened, i.e. phenolics were accumulated in the medium, the activity of polyphenol oxidase (PPO) was increased quickly and secondary wall of cells was thickened. The activity of L _phenylalanine ammonia lyase (PAL), the critical enzyme of the phenylpropanoid pathway, was increased promptly 1 h after elicitation. The rate of taxol production was improved sharply and the maximal taxol concentration at 72 h was six times that of control. Appearance of cell apoptosis was accompanied with the highest concentration of taxol in suspension cultures.
基金National Natural Science Foundation of China(Grant No.39970005)for its financial support
文摘Yeast strain Y68 producing high level of pullulan was isolated from the phyton collected in Toulouse, France. This strain was identified to be Rhodotorula bacarum by BIOLOG analysis. This is the first report that pullulan was produced by Rhodotorula bacarum. The optimal medium (g L -1) for pullulan production by this strain was 80 glucose, 20 soybean cake hydrolysate, 5 K 2HPO 4, 1 NaCl, 0.2 MgSO 4·7H 2O, 0.6 (NH 4) 2SO 4, pH 7.0. Under this condition, 54 g L -1 pullulan was produced within 60 h at 30 ℃. Pullulan is a better starting material for producing marine prodrugs.
基金Supported by the National Key Basic Research Development Program of China, 973 Program, No. 2004CB117505
文摘AIM： To isolate and identify the soybean conglycinin peptides that selectively stimulates the growth of bifidobacteria in vitro, and to investigate the effect of soybean conglycinin peptides on intestinal ecosystem in vivo. METHODS： Soybean conglycinin was purified from soybean seeds by gel filtration （Sepharose-CL-6B）. These proteins were submitted to hydrolysis by pepsin. Several growth-stimulating peptides for bifidobacteria were isolated chromatographically from pepsin hydrolysis of soybean conglycinin and identified by means of matrixassisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. Parallel to in vitro study, in vivo experiments with soybean conglycinin peptides were performed in mice. Ninety male KM mice were randomly assigned into five groups of 16 mice each, and each group was administered for 21d intragastrically with physiological saline （control）, conglycinin, pepsin-treated conglycinin （PTC）, the most active fraction which isolated from pepsin-treated conglycinin （P2-PTC） and HCl-full hydrolysis of conglycinin （HCl-FHC）, respectively. Intestinal microflora were evaluated by standard microbiologic methods and biochemical assays of cecal content samples after treatment. RESULTS： The results showed that the peptides which were isolated from soybean conglycinin could stimulate the growth of bifidobacteria in vitro, and the molecular mass of purified peptides with MALDI-TOF-MS ranged from 693.32 to 1829.55. Compared with control group, in vivo experiments showed that P2-PTC group decreased cecal pH （7.08±0.08 vs7.21±0.09, P〈0.05） and enterococcicounts （5.38±0.26 log10CFU/g vs 5.78±0.19 log10CFU/g, P〈0.05）, significantly increased sIgA level （172.08±35.40 ng/g vs 118.27±33.93 ng/g, P〈0.01） and β-galactosidase activity （1.28±0.23 U/g vs 1.82±0.58 U/g, P〈0.05） CONCLUSION： The results have shown that conglycinin is good source for enzyme-mediated production of peptides which stimulate the growth of bifidobacteria. These peptides are inactive within the sequence of the parent protein but can be released during enzymatic hydrolysis, and in vivo experiments demonstrate that conglycinin peptides may be beneficial for improving gastrointestinal health.
文摘Soy protein isolate/carboxymethyl chitosan （SPI/CMCH） blended films incorporated with glycerol were prepared using solution casting to investigate the effects of the SPI and CMCH ratios （100：0, 88：12, 67：33, 50：50, 33：67, 12：88, 0：100） on the water sorption isotherm. The moisture sorption isotherm of the SPI/CMCH blended films was determined using various relative humidity＇s （16%, 35%, 55% and 76% RH） at 25 ± 1℃. The isotherms showed that the equilibrium moisture content （EMC） of the films increased with increasing CMCH content and the EMC value sharply increased above aw = 0.55. Understanding of sorption isotherms is important for prediction of moisture sorption properties of films via moisture sorption empirical models. The Guggenheim-Oswin, Brunauer-Emmett-Teller （BET）, and Anderson-de Boer （GAB） sorption model predictions were tested against the experimental data. The root mean square （RMS） values from the Oswin, BET, and GAB models respectively ranged from 698.54 to 1,557.54, 38.85 to 58.30, and 52.52 to 95.95. Therefore, the BET model was found to be the best-fit model for SPI/CMCH blended films at 25 ± 1 ℃.
基金We thank for the funding support from the Program of Survey of Chinese Medicines of China(No.[2017]66).
文摘Objective To explore the pharmacological mechanism of active saponin compounds of Tuchao Baibiandouren(Lablab Semen Album fried with earth,TCBBDR)in treating type 2 diabetes(T2DM)using UHPLC-Q-Exactive Orbitrap MS and network pharmacology.Methods UHPLC-Q-Exactive Orbitrap MS was used for a qualitative analysis of saponin compounds in TCBBDR.PharmMapper and CTD were used to screen drug active compounds and disease targets,and an active compound-target network was constructed via Cytoscape 3.8.0.Molecular docking was applied with the drug active compounds and key targets using AutoDock Vina 1.1.2,and a trajectory for the molecular dynamics simulation was completed by GROMACS 2019-3.Results Sixteen saponin compounds were identified from TCBBDR,along with 292 saponin compoud targets and 792 T2DM targets.Through Venn analysis of target saponin constituents and T2DM related targets,a total of 91 intersection targets were screened out in the treatment of T2DM with saponin.The mean values of degree,betweenness centrality and closeness centrality were taken as the thresholds to screen out 22 key genes,among which 4 key proteins namely MAPK1,IGF1 EGFR,PIK3R1 were selected in the top 10 key genes.On this basis,the saponin active constituent-target-signaling pathway network was established.The Gene Ontology(GO)enrichment analysis showed that the related biological modules included activity of steroid hormone receptor,steroid binding,and insulin receptor binding,etc.;the related signaling pathways were EGFR,PI3K-Akt and MAPK,etc.;regulating signaling pathways like MAPK could induce the proliferation,inhibition and apoptosis of pancreaticβcells,increase the quantity of pancreaticβcells,improve the functions of pancreaticβcells and stimulate the insulin secretion.Docking experiment analysis showed that all selected saponin compounds could enter the active sites of targets and form 3–14 hydrogen bonds with residues of the active sites.Moreover,van der Waals forces were present between chemical compounds and active sites.By combining the docking binding energy,we determined that the chemical compounds showed strong binding energy to the targets.Conclusion TCBBDR exerts therapeutic effects on diabetes through multi-compound and multi-target collaboration.Specifically,saponin components mediate pathways including inflammatory reaction and signal transduction to treat T2DM by regulating several key proteins that interact with EGFR and a series of signaling pathways related to disease development.
基金financially supported by grant from the National Natural Science Foundation of China(No.31072219)
文摘Soybean stachyose (SBS) and phytic acid (PA) are anti-nutritional factors (ANF) which have deleterious effects on the growth and digestibility in fish. The present research studied the effects of dietary SBS and PA on the expression of three serine protease genes in the liver of Japanese flounder (Paralichthys olivaceus). These genes are trypsinogen 1 (poTRY), elastase 1 (poEL) and chymotrypsinogen 1 (poCTRY). Eight artificial diets with graded levels of supplemented ANFs were formulated to 4 levels of SBS (0.00, 0.40, 0.80 and 1.50%), 4 levels of PA (0.00, 0.20, 0.40 and 0.80), respectively.Japanese flounder (initial weight 2.45 g ± 0.01 g) were fed with these diets for 10 weeks with three replications per treatment. At the end of 10 weeks, supplementation of 0.40% of dietary SBS or PA significantly increased the gene expression of poTRY and poCTRY (P<0.05). The same level of dietary SBS significantly decreased the gene expression of poEL. In comparison with the control group (ANF-free),dietary PA (0.2% and 0.8%) significantly decreased the gene expression of poTRY, poCTRY and poEL (P<0.05). However, excessive supplement of dietary SBS (1.5%) has no significant effects on these gene expressions (P>0.05). These results suggested that dietary SBS and dietary PA could directly affect the serine protease genes at the transcriptional level in Japanese flounder, and these genes'expression was more sensitive to dietary PA than to SBS under the current experimental conditions.
文摘Using Undaria pinnatifida juice and soya bean as main material, glucono delta lactone （GDL） as coagulant, we produced light green and very nourishing Undaria pinnatifida bean curd. Using the single factor experiment to determine the quantity and concentration of Undaria pinnatifida juice milk, process conditions of coagulation is determined by three factors three levels experiment, the best formulation of Undaria pinnatifida tofu is determined by test of four factors and three levels, process conditions are： the addition of Undaria pinnatifida is 20%, soya-bean milk concentration is1：4.5, the amount of GDL is0.30%, solidification time is 15min, solidification temperature is 95 ℃.
