This study aimed to investigate the influence of pH,heat and enzymatic treatments on the activity of antibacterial substance in MRS,skim soy milk and bovine milk media fermented by a potential probiotic Lactobacillus ...This study aimed to investigate the influence of pH,heat and enzymatic treatments on the activity of antibacterial substance in MRS,skim soy milk and bovine milk media fermented by a potential probiotic Lactobacillus fermentum F6.The antibacterial activity of the culture supernatant of L.fermentum F6 was tested against a wide range of Gram-positive and Gram-negative pathogenic bacteria including Staphylococcus aureus,Escherichia coli,Listeria monocytogenes,Salmonella typhimurium,and Shigella flexneri.Different antibacterial activities were detected in MRS and milk but not in soy milk.We presumed that three kinds of probable components including organic acids,heat-stable and heat-labile proteinaceous compounds were involved in antibacterial activity of fermented MRS and milk.The influence of acids on antibacterial activity was pH-dependent and this effect multiplied with thermal treatments seemed complex.Data analyses showed various significant differences of antibacterial activity among five pathogens were observed in pH and heat treatments.The untreated fermented milk showed higher inhibitory activity to Gram-positive pathogenic bacteria than Gram-negative bacteria （P 〈 0.05）,indicating antibacterial substances of fermented milk are greatly different from fermented MRS.An accurate stimulated gastrointestinal transit showed antibacterial substances would have no influence on intestinal flora.Acidic range of antibacterial substances from pH 2.0 to 6.0 can be potentially used as food biopreservatives and alternative therapeutics.展开更多
Dynamic manipulation of enzymatic activity is a challenging task for applications in chemical and pharmaceutical industries due to the difficult modification and variable conformation of various enzymes.Here, we repor...Dynamic manipulation of enzymatic activity is a challenging task for applications in chemical and pharmaceutical industries due to the difficult modification and variable conformation of various enzymes.Here, we report a new strategy for reversible dynamic modulation of enzymatic activity by near-infrared light-induced photothermal conversion based on polyphenol-functionalized liquid metal nanodroplets(LM). The metal-phenolic nanocoating not only provides colloidal stability of LM nanodroplets but also generates nanointerfaces for the assembly of various enzymes on the LM nanodroplets. Upon near infrared(NIR) irradiation, the localized microenvironmental heating through photothermal effect of the LM nanodroplets allows tailoring the enzymatic activity without affecting the bulk temperature. A library of functional enzymes, including proteinase K, glucoamylase, glucose oxidase, and Bst DNA polymerase, is integrated to perform a reversible control and enhanced activities even after five times of cycles, demonstrating great potential in bacterial fermentation, bacteriostasis, and target gene amplification.展开更多
The effect of superheated steam(SHS)treatment on the quality characteristics of rape bee pollen were studied,and the efficiency of inactivation and inhibition of lipid oxidation were analyzed to investigate the differ...The effect of superheated steam(SHS)treatment on the quality characteristics of rape bee pollen were studied,and the efficiency of inactivation and inhibition of lipid oxidation were analyzed to investigate the differences between SHS and cobalt-60 isotope(^(60)Co)radiation treatment.The number of total plate count(TPC)and mold colonies(MC)remained within the limits of the standards after SHS treatment at 140℃for 2 min.Neither TPC nor MC were detected after^(60)Co irradiation.Peroxidase(POD)and polyphenol oxidase(PPO)activities significantly decreased with increasing temperature and duration of SHS,while^(60)Co radiation completely inactivated PPO.Compared to^(60)Co radiation,SHS treatment inhibited the deterioration of rape bee pollen by avoiding hydroperoxide production and lipid oxidation due to lack of oxygen.These results suggested SHS under 140℃for 2 min was the most suitable to inactivate the microorganisms and enzymes in rape bee pollen with minimal lipid oxidation.展开更多
Film mulching system is a widely employed agricultural practice worldwide. However, the effects of different planting and mulching patterns on soil nutrient content and enzymatic activity have not been well documented...Film mulching system is a widely employed agricultural practice worldwide. However, the effects of different planting and mulching patterns on soil nutrient content and enzymatic activity have not been well documented. In this study, we examined the impact of four planting and mulching patterns（including control, flat planting without mulching; M1, flat planting with film mulching; M2, ridge-furrow planting with film mulching on both ridges and furrows; and M3, ridge-furrow planting with film mulching on continuous ridges） on the seed yield of winter oilseed rape, soil moisture, soil temperature, soil organic carbon（SOC） content, soil nutrient content, and soil enzymatic activity over three growing seasons from 2012 to 2015 in a winter oilseed rape field in the semi-arid area of Northwest China. Seed yield of winter oilseed rape, soil moisture, soil temperature, enzymatic activities, and contents of nitrate-nitrogen, available phosphorus, and available potassium were all significantly higher in mulching treatments（M1, M2 and M3） than in control treatment over the three growing seasons, whereas SOC content was significantly lower in mulching treatments than in control treatment during 2013–2014 and 2014–2015. Among the three mulching treatments（M1, M2 and M3）, the M3 treatment showed consistently higher seed yield, SOC content, nutrient contents, and enzymatic activities than the other two treatments. Seed yield of winter oilseed rape was 41.1% and 15.0% higher in M3 than in M1 and M2, respectively. SOC content and soil enzymatic activities in the top 0–20 cm soil layers and nitrate-nitrogen content in the top 0–30 cm soil layers were all significantly higher in M3 than in M1 and M2. Therefore, we advise the ridge-furrow planting with film mulching on continuous ridges（i.e., M3） as an efficient planting and mulching pattern for sustainably improving the seed yield of winter oilseed rape and preserving soil fertility in the semi-arid area of Northwest China.展开更多
Liquidambar formosana Hance is an attractive landscape tree species because its leaves gradually change from green to red,purple or orange in autumn.In this study,the red variety of L.formosana was used to establish a...Liquidambar formosana Hance is an attractive landscape tree species because its leaves gradually change from green to red,purple or orange in autumn.In this study,the red variety of L.formosana was used to establish a quantitative model of leaf color.Physiological changes in leaf color,pigment levels,enzyme activity,photosynthetic fluorescence characteristics and chloroplast ultrastructure were monitored.The relationship between leaf color and physiological structure indices was quantitatively analyzed to systematically explore the mechanisms behind leaf color.Our data showed that with a decrease in external temperatures,chloroplast numbers and sizes gradually decreased,thylakoid membranes became distorted,and chlorophyll synthesis was blocked and gradually decreased.As a result,chloroplast membranes could not be biosynthesized normally;net photosynthesis,maximum and actual photochemical efficiency,and rate of electron transfer decreased rapidly.Excess light energy caused leaf photoinhibition.With intensification of photoinhibition,leaves protected themselves using two mechanisms.In the first,anthocyanin synthesis was promoted by increasing chalcone isomerase and flavonoid glycosyltransferase activities and soluble sugar content so as to increase anthocyanin to filter light and eliminate reactive oxygen species to reduce photoinhibition.In the second,excessive light energy was consumed in the form of heat energy by increasing the non-photochemical quenching coefficient.These processes tuned the leaves red.展开更多
Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 in...Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 pmol.mg-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.展开更多
The application of enzymatic extracts and conidia of Beauveria bassiana in Metamasius spinolae and Cyclocephala lunulata was evaluated. The variables were mortality and time of death. In M. spinolae, mortality with ex...The application of enzymatic extracts and conidia of Beauveria bassiana in Metamasius spinolae and Cyclocephala lunulata was evaluated. The variables were mortality and time of death. In M. spinolae, mortality with extracts 29%, conidia 27% and the combination of both 31%, all had a time of death of four days. Although with different symptoms, used enzymatic extracts: contraction and softening of the joints;by conidia: mycelium in the joints;in the combination of conidia and enzymatic extracts: abundant aerial mycelium. In C. lunulata, 100% mortality in all treatments;Time of death: enzymatic extracts and extracts with conidia 1.2 days;conidia 2.8 days. Symptoms were different, enzymatic extracts: melanization and internal tissue lysis;enzymatic extract and conidia: mycelium emerged and melanization;conidia: mycelium emerged. Enzymatic extracts showed insecticidal activity in M. spinolae and C. lunulata. These results suggest the potential of enzymatic extracts as biocontrol agents to improve the use of entomopathogenic fungi.展开更多
C-Oligosaccharides are rare in nature and possess diverse bioactivities.However,their chemical synthesis faces many challenges.In this work,enzymatic introduction of C-linked sugar chains to target aglycones was succe...C-Oligosaccharides are rare in nature and possess diverse bioactivities.However,their chemical synthesis faces many challenges.In this work,enzymatic introduction of C-linked sugar chains to target aglycones was successfully achieved by multi-enzymatic cascade reactions.A C-glycosyltransferase from Aloe barbadensis was employed to introduce the first C-linked glucose and then a cyclomaltodextrin glucanotransferase from Bacillus licheniformis was used to extend the sugar chain.A total of twenty C-oligosaccharides with 2-6 sugars were synthesized from scale-up reactions and exhibited good water solubility and sodium-dependent glucose transporter 2(SGLT2)inhibitory activity.Furthermore,a glucoamylase was used to control the length of the sugar chain and the C-maltosides were efficiently synthesized.These findings not only expanded the structural diversity of C-oligosaccharides,but also provided a strategy for the modification of C-glycoside drugs to improve the druggability.展开更多
The H-K-ATPase (HKA), a potassium-dependent proton transporter in the outer medullary collecting duct (OMCD) plays an important role in acid-base homeostasis. The OMCD contains two HKA isoforms;gastric (HKAα1), domin...The H-K-ATPase (HKA), a potassium-dependent proton transporter in the outer medullary collecting duct (OMCD) plays an important role in acid-base homeostasis. The OMCD contains two HKA isoforms;gastric (HKAα1), dominant under normal dietary conditions (ND), and colonic (HKAα2), induced under a K-free diet (KD). The enzymatic activity (EA) of HKA in the OMCD is incompletely understood. The focus of the present study is elucidating the EA of the HKA in HKAα1 and HKAα2 knockout (KO) mice under ND and KD. KO mice were subjected to ND or KD for 10 days. Ten OMCD tubules were extracted, half placed in potassium-free media (Solution 2), half in potassium-containing media (Solution 3). Fluorescence measurements are based on the hydrolysis of ATP to ADP, coupled with the oxidation of NADH. ADP is determined by a decrease in NADH fluorescence. In K presence, NADH fluorescence of HKAα1 KO mice read 13.5 ± 0.7 ppm for ND and 10.3 ± 0.2 ppm for KD, indicating stimulation of the colonic isoform. HKAα2 KO mice averaged 6.8 ± 0.3 ppm for ND and 5.4 ± 0.3 ppm for KD in solution 2 (p p α2 isoform. A significant difference in ATP production in HKAα2 KO mice is likely due to enhanced EA of H-ATPase under potassium depletion.展开更多
Enzymatic activities of beta-gincosidase （ β-GLC ）, （ LAP）, and alkaline phosphatase （ APA）, corresponding to nutrient eliminations, and the microbial community structures were analyzed in an anaerobic-anoxic-a...Enzymatic activities of beta-gincosidase （ β-GLC ）, （ LAP）, and alkaline phosphatase （ APA）, corresponding to nutrient eliminations, and the microbial community structures were analyzed in an anaerobic-anoxic-aerobic reactor system. Results showed that most activity of β-GLC （64.2 μmoi/（ L · h ）） associated with the largest fraction of small- molecular-weight carbohydrates was found in the aerobic reactor, indicating the existence of coupled hydrolysis-uptake mechanism in the aerobic bacteria. Similar activities of LAP presented in the anoxic and aerobic environments, whose increases accompanied by increments in nitrogen uptake rates greatly accelerated the processes of aerobic nitrification and anoxic denitrification. The highest APA activity displayed in the anaerobic reactor, however, dephosphorization performance was mainly achieved under aerobic condition. Microbial community fingerprints generated by polymerase chain reaction amplification and denaturing gradient gel dectrophoresls （ PCR-DGGE ） revealed that Proteobacterium, Actinobacterium, and Nitrosplra were the predominant classes in the activated sindge and there was no evidence of community variations among each function reactor in the system with biomass recycling.展开更多
A fraction of cobra （Naja naja atra） venom has been discovered to have protective effect on rat cultural cerebellar granule neurons （CGNs） from apoptosis induced by removing serum and reducing the extracellular po...A fraction of cobra （Naja naja atra） venom has been discovered to have protective effect on rat cultural cerebellar granule neurons （CGNs） from apoptosis induced by removing serum and reducing the extracellular potassium concentration from 25 to 5 mM. This component has been purified and identified as secreted phospholipase A2 （cobra sPLA2）. In order to study the relationship between the protection on CGNs and enzymatic activity of phospholipase A2, CGNs stained by Hoechst 33258 were quantified to determine survival rate under the fluorescence microscopy; the protective potencies on apoptosis of cultural CGNs were compared among cobra sPLA2, the cobra sPLA2 modified in carboxylate groups with water soluble carbodiimide and semicarbazide, and the heated cobra sPLA2 at 80℃ for 30 rain. The results showed that the CGN survival rate was unaffected significantly both in modified cobra sPLAz whose enzymatic activity of PLA2 had decreased by 80%, and in cobra sPLA2 adding 7, 7-Dimethyleicosadienoic acid, an inhibitor of sPLA2 at concentration of 10-fold IC50; contrary, the neuronal survival rate fell about 60% in heated cobra sPLA2, although its PLA2 activity only decreased by 10%. The protection on CGNs were also found in some of sPLA2s derived from venoms of bee, Naja naja mossambica, Crotalus atroxalso and Vipera Ammodytes Ammodytes but could not be found in other sPLA2s from bovine pancreas and Streptomyces violaceoruber. Above results suggest that the protection on CGNs of cobra sPLA2 is independent of its enzymatic activity.展开更多
Phospholipase A1(PLA1)is a kind of specific phospholipid hydrolase widely used in food,medical,textile.However,limitations in its expression and enzymatic activity have prompted the investigation of the phospholipase-...Phospholipase A1(PLA1)is a kind of specific phospholipid hydrolase widely used in food,medical,textile.However,limitations in its expression and enzymatic activity have prompted the investigation of the phospholipase-assisting protein PlaS.In this study,we elucidate the role of PlaS in enhancing the expression and activity of PlaA1 through N-terminal truncation.Our research demonstrates that truncating the N-terminal region of PlaS effectively overcomes its inhibitory effect on host cells,resulting in improved cell growth and increased protein solubility of the protein.The yeast two-hybrid assay confirms the interaction between PlaA1 and N-terminal truncated PlaS(ΔN27 PlaS),highlighting their binding capabilities.Furthermore,in vitro studies using Biacore analysis reveal a concentration-dependent and specific binding between PlaA1 andΔN27 PlaS,exhibiting high affinity.Molecular docking analysis provides insights into the hydrogen bond interactions betweenΔN27 PlaS and PlaA1,identifying key amino acid residues crucial for their binding.Finally,the enzyme activity of PLA1 was boost to 8.4 U/mL by orthogonal test.Study significantly contributes to the understanding of the interaction mechanism between PlaS and PlaA1,offering potential strategies for enhancing PlaA1 activity through protein engineering approaches.展开更多
Saponins can be potential candidates for the development of safe biopesticides,due to their widely acknowledged insecticidal,fungicidal and nematicidal activity,but information on their effects on soil biological prop...Saponins can be potential candidates for the development of safe biopesticides,due to their widely acknowledged insecticidal,fungicidal and nematicidal activity,but information on their effects on soil biological properties is still limited.This study aimed to investigate the short-term fate of saponins from Medicago sativa in soil and their dose-effect relationship with microbial biomass and selected enzyme activities in soils with different origin,physical and chemical properties.Microbial degradation of total saponins ranged from 46%to 91%,according to soil characteristics,within 28 days from their incorporation into the soil.Both saponin glycosidic chains and triterpenic aglycones were also microbially degraded,though by dynamics changing among the different soils.In all soils,M.sativa saponins significantly reduced microbial biomass at rates of 10 and 20 mg saponin mixture per g of soil.Microbial enzymatic activities were less affected as indicating an adaptive response of soil microbial communities to the presence of saponins.展开更多
The biochemical response of Coffea arabica var.Borbon to chitosan and chitosan oligosaccharides(COS)was evaluated in one-year-old plants under greenhouse conditions.COS solutions were synthesized through chemical and ...The biochemical response of Coffea arabica var.Borbon to chitosan and chitosan oligosaccharides(COS)was evaluated in one-year-old plants under greenhouse conditions.COS solutions were synthesized through chemical and physical hydrolysis using acetic acid,hydrogen peroxide,and microwave irradiation.The obtained COS had an average molecular weight(Mw)of 3549.90±0.33 Daltons(Da),a deacetylation degree(DD)of 76.64±1.12%,and a polymerization degree(PD)of 18.91±0.0018.Solutions of chitosan and COS were applied to C.arabica var.Borbon at concentrations of 0.25,0.5,and 1 wt%.The experimental design was conducted using a completely randomized design with four replications.The biochemical responses assessed included soluble protein content,phenylalanine ammonia-lyase(PAL),chitinase,β-1,3-glucanase,peroxidase,catalase,and chlorophyll fluorescence.The application of COS demonstrated significant differences(α=0.05)in protein concentration,with the activity ofβ-1,3-glucanase,chitinase,and catalase being 1.5,7.5,and 3.9 times higher,respectively,while showing similar behavior to chitosan in PAL activity,both up to 4.4 times higher than the distilled water control and lower than chitosan in peroxidase activity.Treatments with chitosan yielded a higher photochemical efficiency of Photosystem II(PSII).The application of COS suggests a viable foliar alternative to active plant defense mechanisms without the risk of phytotoxicity.展开更多
Bioremediation has gained significant attention due to its potential to remove azo dyes.However,the challenges microor-ganisms face in surviving when azo dyes are the sole carbon source limit its widespread applicatio...Bioremediation has gained significant attention due to its potential to remove azo dyes.However,the challenges microor-ganisms face in surviving when azo dyes are the sole carbon source limit its widespread application.This study aimed to improve the biodegradation of azo dyes by utilizing Baijiu distiller’s grains leachate(BDGL)as a co-substrate.The experimental results demon-strated that BDGL significantly enhanced Providencia rettgeri’s ability to degrade the model pollutant Acid Black 210(AB210),achieving a decolorization efficiency of 94.5%.This may be attributed to the nutrient-rich composition of BDGL,which includes ethanol and protein,providing a favorable substrate for bacterial growth and activity.The higher biomass and increased activities of azoreductase and quinone oxidoreductase in the BDGL group further supported these findings.Additionally,this method demonstra-ted broad-spectrum degradation of azo dyes(Direct Red 5B,Acid Red 73,and Congo Red)with different structures,highlighting its potential applicability.Metabolite assays combined with transcriptomics analyses revealed that the expression of functional genes re-lated to redox reactions,azo bond cleavage,and hydrolysis increased under the co-metabolic conditions of BDGL,resulting in stronger reducing power that further mineralized the dye into smaller metabolites.Our study offers a practical strategy for the simulta-neous treatment of dye-containing wastewater and Baijiu distiller’s grains,with significant environmental and industrial applications.展开更多
基金the National Nature Science Foundation of China (30660135, 30800861)the National High-Tech R&D Program of China (863 Program, 2006AA10Z345, 2007AA10Z353)the Ear Marked Fund for Modern Agro-Industry Technology Research System, China (NCET-06-0269)
文摘This study aimed to investigate the influence of pH,heat and enzymatic treatments on the activity of antibacterial substance in MRS,skim soy milk and bovine milk media fermented by a potential probiotic Lactobacillus fermentum F6.The antibacterial activity of the culture supernatant of L.fermentum F6 was tested against a wide range of Gram-positive and Gram-negative pathogenic bacteria including Staphylococcus aureus,Escherichia coli,Listeria monocytogenes,Salmonella typhimurium,and Shigella flexneri.Different antibacterial activities were detected in MRS and milk but not in soy milk.We presumed that three kinds of probable components including organic acids,heat-stable and heat-labile proteinaceous compounds were involved in antibacterial activity of fermented MRS and milk.The influence of acids on antibacterial activity was pH-dependent and this effect multiplied with thermal treatments seemed complex.Data analyses showed various significant differences of antibacterial activity among five pathogens were observed in pH and heat treatments.The untreated fermented milk showed higher inhibitory activity to Gram-positive pathogenic bacteria than Gram-negative bacteria （P 〈 0.05）,indicating antibacterial substances of fermented milk are greatly different from fermented MRS.An accurate stimulated gastrointestinal transit showed antibacterial substances would have no influence on intestinal flora.Acidic range of antibacterial substances from pH 2.0 to 6.0 can be potentially used as food biopreservatives and alternative therapeutics.
基金financial support from the National Talents Program, National Natural Science Foundation of China (Nos. 22178233, 22108181)Talents Program of Sichuan Province, Double First-Class University Plan of Sichuan University, State Key Laboratory of Polymer Materials Engineering (No. sklpme 2020-03-01)the Sichuan Province Postdoctoral Special Funding。
文摘Dynamic manipulation of enzymatic activity is a challenging task for applications in chemical and pharmaceutical industries due to the difficult modification and variable conformation of various enzymes.Here, we report a new strategy for reversible dynamic modulation of enzymatic activity by near-infrared light-induced photothermal conversion based on polyphenol-functionalized liquid metal nanodroplets(LM). The metal-phenolic nanocoating not only provides colloidal stability of LM nanodroplets but also generates nanointerfaces for the assembly of various enzymes on the LM nanodroplets. Upon near infrared(NIR) irradiation, the localized microenvironmental heating through photothermal effect of the LM nanodroplets allows tailoring the enzymatic activity without affecting the bulk temperature. A library of functional enzymes, including proteinase K, glucoamylase, glucose oxidase, and Bst DNA polymerase, is integrated to perform a reversible control and enhanced activities even after five times of cycles, demonstrating great potential in bacterial fermentation, bacteriostasis, and target gene amplification.
基金supported by National Natural Science Foundations of China(32472396,31871861 and 31501548)The Apicultural Industry Technology System(NCYTI-43-KXJ17)The Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences(CAAS-ASTIP-2015-IAR)。
文摘The effect of superheated steam(SHS)treatment on the quality characteristics of rape bee pollen were studied,and the efficiency of inactivation and inhibition of lipid oxidation were analyzed to investigate the differences between SHS and cobalt-60 isotope(^(60)Co)radiation treatment.The number of total plate count(TPC)and mold colonies(MC)remained within the limits of the standards after SHS treatment at 140℃for 2 min.Neither TPC nor MC were detected after^(60)Co irradiation.Peroxidase(POD)and polyphenol oxidase(PPO)activities significantly decreased with increasing temperature and duration of SHS,while^(60)Co radiation completely inactivated PPO.Compared to^(60)Co radiation,SHS treatment inhibited the deterioration of rape bee pollen by avoiding hydroperoxide production and lipid oxidation due to lack of oxygen.These results suggested SHS under 140℃for 2 min was the most suitable to inactivate the microorganisms and enzymes in rape bee pollen with minimal lipid oxidation.
基金supported by the Special Fund for Agro-scientific Research in the Public Interest,China (201503125,201503105)the National High Technology Research and Development Program of China (2011AA100504)
文摘Film mulching system is a widely employed agricultural practice worldwide. However, the effects of different planting and mulching patterns on soil nutrient content and enzymatic activity have not been well documented. In this study, we examined the impact of four planting and mulching patterns（including control, flat planting without mulching; M1, flat planting with film mulching; M2, ridge-furrow planting with film mulching on both ridges and furrows; and M3, ridge-furrow planting with film mulching on continuous ridges） on the seed yield of winter oilseed rape, soil moisture, soil temperature, soil organic carbon（SOC） content, soil nutrient content, and soil enzymatic activity over three growing seasons from 2012 to 2015 in a winter oilseed rape field in the semi-arid area of Northwest China. Seed yield of winter oilseed rape, soil moisture, soil temperature, enzymatic activities, and contents of nitrate-nitrogen, available phosphorus, and available potassium were all significantly higher in mulching treatments（M1, M2 and M3） than in control treatment over the three growing seasons, whereas SOC content was significantly lower in mulching treatments than in control treatment during 2013–2014 and 2014–2015. Among the three mulching treatments（M1, M2 and M3）, the M3 treatment showed consistently higher seed yield, SOC content, nutrient contents, and enzymatic activities than the other two treatments. Seed yield of winter oilseed rape was 41.1% and 15.0% higher in M3 than in M1 and M2, respectively. SOC content and soil enzymatic activities in the top 0–20 cm soil layers and nitrate-nitrogen content in the top 0–30 cm soil layers were all significantly higher in M3 than in M1 and M2. Therefore, we advise the ridge-furrow planting with film mulching on continuous ridges（i.e., M3） as an efficient planting and mulching pattern for sustainably improving the seed yield of winter oilseed rape and preserving soil fertility in the semi-arid area of Northwest China.
基金This work was supported by the Fundamental Research Funds for Guangxi Zhuang Autonomous Region Forestry Research Institute(No.Linke 201809)Guangxi Forestry Science and Technology Promotion Demonstration Project(No.26 Guilin Research 2021).
文摘Liquidambar formosana Hance is an attractive landscape tree species because its leaves gradually change from green to red,purple or orange in autumn.In this study,the red variety of L.formosana was used to establish a quantitative model of leaf color.Physiological changes in leaf color,pigment levels,enzyme activity,photosynthetic fluorescence characteristics and chloroplast ultrastructure were monitored.The relationship between leaf color and physiological structure indices was quantitatively analyzed to systematically explore the mechanisms behind leaf color.Our data showed that with a decrease in external temperatures,chloroplast numbers and sizes gradually decreased,thylakoid membranes became distorted,and chlorophyll synthesis was blocked and gradually decreased.As a result,chloroplast membranes could not be biosynthesized normally;net photosynthesis,maximum and actual photochemical efficiency,and rate of electron transfer decreased rapidly.Excess light energy caused leaf photoinhibition.With intensification of photoinhibition,leaves protected themselves using two mechanisms.In the first,anthocyanin synthesis was promoted by increasing chalcone isomerase and flavonoid glycosyltransferase activities and soluble sugar content so as to increase anthocyanin to filter light and eliminate reactive oxygen species to reduce photoinhibition.In the second,excessive light energy was consumed in the form of heat energy by increasing the non-photochemical quenching coefficient.These processes tuned the leaves red.
基金supported by grants from the Scientific Research Foundation for Returned Scholars of the Ministry of Education of ChinaPre-research Foundation of Wuhan University,China(No.301270050)
文摘Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 pmol.mg-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.
文摘The application of enzymatic extracts and conidia of Beauveria bassiana in Metamasius spinolae and Cyclocephala lunulata was evaluated. The variables were mortality and time of death. In M. spinolae, mortality with extracts 29%, conidia 27% and the combination of both 31%, all had a time of death of four days. Although with different symptoms, used enzymatic extracts: contraction and softening of the joints;by conidia: mycelium in the joints;in the combination of conidia and enzymatic extracts: abundant aerial mycelium. In C. lunulata, 100% mortality in all treatments;Time of death: enzymatic extracts and extracts with conidia 1.2 days;conidia 2.8 days. Symptoms were different, enzymatic extracts: melanization and internal tissue lysis;enzymatic extract and conidia: mycelium emerged and melanization;conidia: mycelium emerged. Enzymatic extracts showed insecticidal activity in M. spinolae and C. lunulata. These results suggest the potential of enzymatic extracts as biocontrol agents to improve the use of entomopathogenic fungi.
基金financially supported by the National Key Research and Development Program of China(No.2020YFA0908000)CAMS Innovation fund for Medical Sciences(No.2021-I2M-1–029)。
文摘C-Oligosaccharides are rare in nature and possess diverse bioactivities.However,their chemical synthesis faces many challenges.In this work,enzymatic introduction of C-linked sugar chains to target aglycones was successfully achieved by multi-enzymatic cascade reactions.A C-glycosyltransferase from Aloe barbadensis was employed to introduce the first C-linked glucose and then a cyclomaltodextrin glucanotransferase from Bacillus licheniformis was used to extend the sugar chain.A total of twenty C-oligosaccharides with 2-6 sugars were synthesized from scale-up reactions and exhibited good water solubility and sodium-dependent glucose transporter 2(SGLT2)inhibitory activity.Furthermore,a glucoamylase was used to control the length of the sugar chain and the C-maltosides were efficiently synthesized.These findings not only expanded the structural diversity of C-oligosaccharides,but also provided a strategy for the modification of C-glycoside drugs to improve the druggability.
文摘The H-K-ATPase (HKA), a potassium-dependent proton transporter in the outer medullary collecting duct (OMCD) plays an important role in acid-base homeostasis. The OMCD contains two HKA isoforms;gastric (HKAα1), dominant under normal dietary conditions (ND), and colonic (HKAα2), induced under a K-free diet (KD). The enzymatic activity (EA) of HKA in the OMCD is incompletely understood. The focus of the present study is elucidating the EA of the HKA in HKAα1 and HKAα2 knockout (KO) mice under ND and KD. KO mice were subjected to ND or KD for 10 days. Ten OMCD tubules were extracted, half placed in potassium-free media (Solution 2), half in potassium-containing media (Solution 3). Fluorescence measurements are based on the hydrolysis of ATP to ADP, coupled with the oxidation of NADH. ADP is determined by a decrease in NADH fluorescence. In K presence, NADH fluorescence of HKAα1 KO mice read 13.5 ± 0.7 ppm for ND and 10.3 ± 0.2 ppm for KD, indicating stimulation of the colonic isoform. HKAα2 KO mice averaged 6.8 ± 0.3 ppm for ND and 5.4 ± 0.3 ppm for KD in solution 2 (p p α2 isoform. A significant difference in ATP production in HKAα2 KO mice is likely due to enhanced EA of H-ATPase under potassium depletion.
文摘Enzymatic activities of beta-gincosidase （ β-GLC ）, （ LAP）, and alkaline phosphatase （ APA）, corresponding to nutrient eliminations, and the microbial community structures were analyzed in an anaerobic-anoxic-aerobic reactor system. Results showed that most activity of β-GLC （64.2 μmoi/（ L · h ）） associated with the largest fraction of small- molecular-weight carbohydrates was found in the aerobic reactor, indicating the existence of coupled hydrolysis-uptake mechanism in the aerobic bacteria. Similar activities of LAP presented in the anoxic and aerobic environments, whose increases accompanied by increments in nitrogen uptake rates greatly accelerated the processes of aerobic nitrification and anoxic denitrification. The highest APA activity displayed in the anaerobic reactor, however, dephosphorization performance was mainly achieved under aerobic condition. Microbial community fingerprints generated by polymerase chain reaction amplification and denaturing gradient gel dectrophoresls （ PCR-DGGE ） revealed that Proteobacterium, Actinobacterium, and Nitrosplra were the predominant classes in the activated sindge and there was no evidence of community variations among each function reactor in the system with biomass recycling.
文摘A fraction of cobra （Naja naja atra） venom has been discovered to have protective effect on rat cultural cerebellar granule neurons （CGNs） from apoptosis induced by removing serum and reducing the extracellular potassium concentration from 25 to 5 mM. This component has been purified and identified as secreted phospholipase A2 （cobra sPLA2）. In order to study the relationship between the protection on CGNs and enzymatic activity of phospholipase A2, CGNs stained by Hoechst 33258 were quantified to determine survival rate under the fluorescence microscopy; the protective potencies on apoptosis of cultural CGNs were compared among cobra sPLA2, the cobra sPLA2 modified in carboxylate groups with water soluble carbodiimide and semicarbazide, and the heated cobra sPLA2 at 80℃ for 30 rain. The results showed that the CGN survival rate was unaffected significantly both in modified cobra sPLAz whose enzymatic activity of PLA2 had decreased by 80%, and in cobra sPLA2 adding 7, 7-Dimethyleicosadienoic acid, an inhibitor of sPLA2 at concentration of 10-fold IC50; contrary, the neuronal survival rate fell about 60% in heated cobra sPLA2, although its PLA2 activity only decreased by 10%. The protection on CGNs were also found in some of sPLA2s derived from venoms of bee, Naja naja mossambica, Crotalus atroxalso and Vipera Ammodytes Ammodytes but could not be found in other sPLA2s from bovine pancreas and Streptomyces violaceoruber. Above results suggest that the protection on CGNs of cobra sPLA2 is independent of its enzymatic activity.
基金financially supported by National Natural Science Foundations of China(No.31471615)Scientific Research Start-up Fund for Introduced Talents of Anhui Polytechnic University(2022YOO068)Natural Science Research Project of Colleges and Universities in Anhui Province(Grant 2022AH050971).
文摘Phospholipase A1(PLA1)is a kind of specific phospholipid hydrolase widely used in food,medical,textile.However,limitations in its expression and enzymatic activity have prompted the investigation of the phospholipase-assisting protein PlaS.In this study,we elucidate the role of PlaS in enhancing the expression and activity of PlaA1 through N-terminal truncation.Our research demonstrates that truncating the N-terminal region of PlaS effectively overcomes its inhibitory effect on host cells,resulting in improved cell growth and increased protein solubility of the protein.The yeast two-hybrid assay confirms the interaction between PlaA1 and N-terminal truncated PlaS(ΔN27 PlaS),highlighting their binding capabilities.Furthermore,in vitro studies using Biacore analysis reveal a concentration-dependent and specific binding between PlaA1 andΔN27 PlaS,exhibiting high affinity.Molecular docking analysis provides insights into the hydrogen bond interactions betweenΔN27 PlaS and PlaA1,identifying key amino acid residues crucial for their binding.Finally,the enzyme activity of PLA1 was boost to 8.4 U/mL by orthogonal test.Study significantly contributes to the understanding of the interaction mechanism between PlaS and PlaA1,offering potential strategies for enhancing PlaA1 activity through protein engineering approaches.
基金supported by the ROP ERDF 2014-2020 Lombardy-Innovation and Competitiveness.
文摘Saponins can be potential candidates for the development of safe biopesticides,due to their widely acknowledged insecticidal,fungicidal and nematicidal activity,but information on their effects on soil biological properties is still limited.This study aimed to investigate the short-term fate of saponins from Medicago sativa in soil and their dose-effect relationship with microbial biomass and selected enzyme activities in soils with different origin,physical and chemical properties.Microbial degradation of total saponins ranged from 46%to 91%,according to soil characteristics,within 28 days from their incorporation into the soil.Both saponin glycosidic chains and triterpenic aglycones were also microbially degraded,though by dynamics changing among the different soils.In all soils,M.sativa saponins significantly reduced microbial biomass at rates of 10 and 20 mg saponin mixture per g of soil.Microbial enzymatic activities were less affected as indicating an adaptive response of soil microbial communities to the presence of saponins.
文摘The biochemical response of Coffea arabica var.Borbon to chitosan and chitosan oligosaccharides(COS)was evaluated in one-year-old plants under greenhouse conditions.COS solutions were synthesized through chemical and physical hydrolysis using acetic acid,hydrogen peroxide,and microwave irradiation.The obtained COS had an average molecular weight(Mw)of 3549.90±0.33 Daltons(Da),a deacetylation degree(DD)of 76.64±1.12%,and a polymerization degree(PD)of 18.91±0.0018.Solutions of chitosan and COS were applied to C.arabica var.Borbon at concentrations of 0.25,0.5,and 1 wt%.The experimental design was conducted using a completely randomized design with four replications.The biochemical responses assessed included soluble protein content,phenylalanine ammonia-lyase(PAL),chitinase,β-1,3-glucanase,peroxidase,catalase,and chlorophyll fluorescence.The application of COS demonstrated significant differences(α=0.05)in protein concentration,with the activity ofβ-1,3-glucanase,chitinase,and catalase being 1.5,7.5,and 3.9 times higher,respectively,while showing similar behavior to chitosan in PAL activity,both up to 4.4 times higher than the distilled water control and lower than chitosan in peroxidase activity.Treatments with chitosan yielded a higher photochemical efficiency of Photosystem II(PSII).The application of COS suggests a viable foliar alternative to active plant defense mechanisms without the risk of phytotoxicity.
基金supported by the National Key Re-search and Development Project of China(No.2023YFC3108400).
文摘Bioremediation has gained significant attention due to its potential to remove azo dyes.However,the challenges microor-ganisms face in surviving when azo dyes are the sole carbon source limit its widespread application.This study aimed to improve the biodegradation of azo dyes by utilizing Baijiu distiller’s grains leachate(BDGL)as a co-substrate.The experimental results demon-strated that BDGL significantly enhanced Providencia rettgeri’s ability to degrade the model pollutant Acid Black 210(AB210),achieving a decolorization efficiency of 94.5%.This may be attributed to the nutrient-rich composition of BDGL,which includes ethanol and protein,providing a favorable substrate for bacterial growth and activity.The higher biomass and increased activities of azoreductase and quinone oxidoreductase in the BDGL group further supported these findings.Additionally,this method demonstra-ted broad-spectrum degradation of azo dyes(Direct Red 5B,Acid Red 73,and Congo Red)with different structures,highlighting its potential applicability.Metabolite assays combined with transcriptomics analyses revealed that the expression of functional genes re-lated to redox reactions,azo bond cleavage,and hydrolysis increased under the co-metabolic conditions of BDGL,resulting in stronger reducing power that further mineralized the dye into smaller metabolites.Our study offers a practical strategy for the simulta-neous treatment of dye-containing wastewater and Baijiu distiller’s grains,with significant environmental and industrial applications.
