Kai Xin San is a Chinese herbal formula composed of Radix Ginseng, Poria, Radix Polygalae and Acorus Tatarinowii Rhizome. It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-...Kai Xin San is a Chinese herbal formula composed of Radix Ginseng, Poria, Radix Polygalae and Acorus Tatarinowii Rhizome. It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β (Aβ) induced cognitive dysfunction and is neuroprotective in vivo, but its precise mechanism remains unclear. Expression of insulin-degrading enzyme (IDE), which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 (200 μM, 5 μL) into the hippocampus and subsequently administered Kai Xin San (0.54 or 1.08 g/kg/d) intragastrically for 21 consecutive days. Hematoxylin eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.展开更多
A strain of Bacillus subtilis strain YB 1, isolated and preserved in our lab., showed a high nicosulfuron-degrading activity. Optimization of culture conditions on production of nicosulfuron-degrading enzyme from Baci...A strain of Bacillus subtilis strain YB 1, isolated and preserved in our lab., showed a high nicosulfuron-degrading activity. Optimization of culture conditions on production of nicosulfuron-degrading enzyme from Bacillus subtilis strain YB 1 was carried out through mono-factor experiments. The characterization of degrading enzyme(s) was studied in this paper. The results showed that B. subtilis YB1 can use nicosulfuron as sole carbon source under aerobic condition. The key enzyme(s) involved in the initial biodegradation of nicosulfuron was localized to extracellular proteins and showed to be induced expressed. Enzyme-specific activity was up to 89.34 U mg-1 at pH 8.0 and 30℃, incubation for 96 h, inoculum 4.5x108 CFU mL-1 in Luria-Bertani liquid medium with nicosulfuron of 40 mg L-1. The maximum degradation rate of extracellular crude enzymes on nicosulfuron was 66% at pH 9.0, 35℃ in the enzymatic reaction system with nicosulfuron of 5 mg L-1. This degrading enzyme(s) was sensitive to high temperature, but kept high activity under alkaline conditions.展开更多
This study was designed to analyze the changes of phytohormone concentrations, calcium fraction, and the activities of degrading enzymes during calcium-inhibited and ethyleneglycol-bis-(β-aminoethyl ether)N, N'-te...This study was designed to analyze the changes of phytohormone concentrations, calcium fraction, and the activities of degrading enzymes during calcium-inhibited and ethyleneglycol-bis-(β-aminoethyl ether)N, N'-tetraacetic acid (EGTA)-induced abscission of tomato pedicel explants. Added calcium caused an increase in the total calcium content within the abscission zone and produced a corresponding reduction (20%) in pedicel explant abscission. As expected, EGTA treatment produced the opposite effect and resulted in a decrease in the total calcium content, while accelerating abscission of pedicel explants. Hormone analysis revealed that indole-3-acetic acid (IAA) concentrations in the abscission zone first decreased and then increased before the occurrence of abscission in all treatments, with the greatest effect produced by addition of EGTA. Similarly, abscisic acid (ABA), and gibberellin (GA1+3) concentrations, and ethylene production were elevated in the abscission zone during the first 16 h before abscission when explants imbibed EGTA. With calcium treatment, the concentrations of ABA, ethylene, and GA1+3 also increased in pedicels throughout the first 16 h following exposure, but the increase was slower and less dramatic than with EGTA. Both cellulase and polygalacturonase were induced in the explants during abscission and the activities were also strengthened by treatment with EGTA. Calciumtreated explants produced lower hydrolysing enzyme activities than controls throughout abscission. Calcium played a role of mediating hormone balance and degrading enzymes activities and affected on abscission.展开更多
[ Objective ] This study aimed to confirm the roles of cell wall degrading enzymes (CWDEs) produced by Botryodiplodia theobromae Pat. in the infec- tion of mango fruits. [ Method] Change of activities of five types ...[ Objective ] This study aimed to confirm the roles of cell wall degrading enzymes (CWDEs) produced by Botryodiplodia theobromae Pat. in the infec- tion of mango fruits. [ Method] Change of activities of five types of CWDEs produced by B. theobromae Pat. were studied under both in vitro culture and inocula- tion conditions, along with the pathogenicity and the ability of producing CWDEs of four post-harvest fangal pathogens(B, theobromae Pat. , Colletotrichum gloeos- porioides Penz. , Phomopsis mangiferae Ahmad and Dothiorella dominicana Pet. et Cif. ) which cause stem-end rot of mangoes. [ Result] B. theobromae Pat. was a- ble to produce polygalacturonase(PG), pectinmethylgalacturonase(PMG), polygalacturonic acid trans-eliminase (PGTE), pectin methyltrans-eliminase (PMTE) and cellulase ( Cx. ) under both in vitro culture and inoculation conditions, of which activities of PG, Cx and PMG were significantly higher in than that in either PGTE or PMTE. Among three primary CWDEs, the peak of activities of PG and Cx appeared earlier and that of PMG occured later. The pathogenicity of B. theo- bromae Pat. was significantly higher than that of any other three pathogens; it is the same with the abilities of producing pectinase. [ Conclusion] This paper pro- vides theoretical bases for further exploring the mechanism of host-pathogen interaction and decreasing the post-harvest loss of mango fruits.展开更多
Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrola...Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE). Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderma. Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique. Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities. Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.展开更多
[ Objective ] This study aimed to obtain the recombinant Pichia yeast strain which can efficiently degrade guar gum. The properties of the recombinant enzyme were studied preliminarily. [ Method ] A positive clone tha...[ Objective ] This study aimed to obtain the recombinant Pichia yeast strain which can efficiently degrade guar gum. The properties of the recombinant enzyme were studied preliminarily. [ Method ] A positive clone that could hydrolyze guar gum was obtained through the construction and functional screening of a soil genome library. Sequence analysis indicated that the 1485-bp clone encodes a 494-amino acid protein with a relative molecular mass of 53 949 kD, containing a cellulose-binding domain. The recombinant plasmid pHBM731 was generated by inserting the optimized target gene into a Pichia pastoris expression vector pHBMg05 that was transformed into three Pichia pastoris strains, GS115, KM71 and SMD1168. The biochemical properties of the enzyme were assessed. [ Result] The cloned galactonumnan (GM)-degrading enzyme was expressed and secreted by Pichia pastoris GSll5. High cell density fermentation was induced in recombi- nant Pichia pastoris at 25 and 28 ~C ; a higher enzyme activity was observed at an induction temperature of 28 ~C. The optimal temperature for the recombinant en- zyme is 60 ~C, and the optimal pH is 6.6. The enzyme activity was 38.61 U under optimal conditions. Over 50% of the enzyme activity was maintained under the optimal conditions after 9 h. Under the optimal conditions, the effect of metal ions on enzyme activity was analyzed. Ca2 + , Fe2 + and Li ~ slightly enhanced enzyme activity, while Mn2+ and Co2+ had little effect. Enzyme activity was modestly suppressed by Mg2~ , K~ and Na+ , but considerably suppressed by Ag2~ and Zn2~ , with Cu2 + showing the strongest inhibitory effects. [ Conclusion] A novel GM-degrading enzyme expressed by soil yeast was cloned, which can potentially be used in industrial applications to obtain eommereially useful guar gum-degradation products.展开更多
Peanuts can be affected by the presence of pathogenic microorganisms. The fungus <i>Thecaphora frezii</i> (<i>T. frezii</i>), which belongs to the taxonomic class Ustilaginomycetes, is the caus...Peanuts can be affected by the presence of pathogenic microorganisms. The fungus <i>Thecaphora frezii</i> (<i>T. frezii</i>), which belongs to the taxonomic class Ustilaginomycetes, is the causal agent of the disease known as “peanut smut”. The life cycle of this fungus includes three stages, namely teliospores, basidiospores and hyphae. In the hyphae stage, infection occurs in the peanut plant, which requires the involvement of some enzymes secreted by the fungus. These include the Plant Cell Wall-Degrading Enzymes (PCWDEs), which degrade various polysaccharides. This study aimed to identify the presence of transcript for enzymes belonging to the PCWDEs from three stages of <i>T. frezii</i>. For this, total RNA was extracted from the three ontogenetic stages of <i>T. frezii</i>. These samples were analyzed using an RNA-Seq approach and some transcripts were quantified using Real Time PCR. The analysis of the data provided by the RNA-Seq of the three <i>T. frezii</i> stages, it was possible to identify some transcripts that could encode enzymes compatible with polysaccharides degradation that are part of the plant cell wall. In <i>T. frezii</i> transcriptome, 40 deduced proteins would be enzymes with functions of PCWDEs were identified. They were divided into 27 glycoside hydrolases;two polysaccharide lyases;three carbohydrate esterases and eight enzymes with auxiliary activities. In addition, the fungal SNF1 gene was identified whose activity could be affected by high glucose level, and indirectly influence the levels of some PCWDEs. The analysis of the PCWDEs could help to understand part of the fungal infection process and possibly find substances that can control its development.展开更多
We used a litterbag method to investigate litter decomposition and related soil degradative enzyme activities across four seasons in a broad-leaved forest and a coniferous forest on Zijin Mountain in sub-tropical Chin...We used a litterbag method to investigate litter decomposition and related soil degradative enzyme activities across four seasons in a broad-leaved forest and a coniferous forest on Zijin Mountain in sub-tropical China. Across four seasons, we quantified litter mass losses, soil pH values, and related soil degradative enzyme activities. Litter decomposition rates differed significantly by season. Litter decomposi- tion rates of broadleaf forest leaves were higher than for coniferous for- ests needles across four seasons, and maximal differences in litter de- composition rates between the two litter types were found in spring.展开更多
Heat and acid treatments were reported to be a promising substitute for SO2 fumigation in color protection of postharvest lychee (Litchi chinensis) fruits, but the mechanism was not clear. In the present study, hot ...Heat and acid treatments were reported to be a promising substitute for SO2 fumigation in color protection of postharvest lychee (Litchi chinensis) fruits, but the mechanism was not clear. In the present study, hot water (70℃) dipping followed by immersion in 2% HC1 (heat-acid) substantially protected the red color of the fruit during storage at 25℃ and inhibited anthocyanin degradation while hot water dipping alone (heat) led to rapidly browning and about 90% loss in anthocyanin content. The pH values in the pericarp of the heat-acid treated fruit dropped to 3.2, while the values maintained around 5.0 in the heat-treated and control fruit. No significantly different pH values were detected among the arils of heat-acid, heat treated and control fruit. Heat-acid treatment dramatically reduced the activities of anthocyanin degradation enzyme (ADE), peroxidase (POD) and polyphenol oxidase in the pericarp. A marked reduction in LcPOD gene expression was also detected in heat-acid treated fruit, in contrast, induction was found in heat treated fruit. The pericarp of heat-acid treated fruit exhibited significantly lower respiration rate but faster water loss than that of the untreated or heat treated fruit. Taken together, heat treatment triggered quick browning and anthocyanin loss in lychee fruit, while heat-acid treatment protected the fruit color by a great reduction in the activities/gene expression of anthocyanin degradation enzymes and acidification of lychee pericarp.展开更多
The mechanism of carotenoid degradation and the changes in the activities of related enzymes in flue-cured tobacco at the leaf-drying stage during the bulk-curing process were studied in order to provide theoretical b...The mechanism of carotenoid degradation and the changes in the activities of related enzymes in flue-cured tobacco at the leaf-drying stage during the bulk-curing process were studied in order to provide theoretical basis for optimization of curing technology. The effect of different rising speeds of temperature on the carotenoid degradation and the related enzymes activities at the color-fixing stage during the bulk curing process was studied by using the electric-heated fluecuring barn designed by Henan Agricultural University, China, based on curing technology with yellowing at low temperature and moderate humidity and leaf drying at moderate humidity. The results showed that the carotenoid degradation components (β-carotene, lutein, neoxanthin, and violaxthin) decreased gradually at the color-fixing stage during the bulk curing process. The carotenoid degradation components viz.,β-carotene, lutein, neoxanthin, and violaxthin at the slow heating curing (T1) were relatively higher than the rapid heating curing (T2) accounting for 10, 2, 32 and 32% respectively, but there were no differences among treatments (P〉 0.05). The effect of different conditions of curing on the activities of enzymes related to carotenoids degradation were significant. The lipoxygenase, phenylalanine ammonialyase, peroxidase, and polyphenol oxidase enzymes had a bidirectional effect on the quality of tobacco leaves and it was beneficial to form more premise matter of aroma based on the higher enzyme activities at the early leaf-drying stage. The slow heating could regulate the change in various enzymes' activities reasonably, making cell redox reaction to reach the dynamic balance and make the degradation of carotenoids adequately. Meanwhile, it could avoid the occurrence of browning reaction and provide foundation for improving the quality of tobacco and optimization of technology for bulk curing and further enhancing aroma.展开更多
The degradation behaviors of Thai Bombyx mori called Samrong and Nanglai silk fibroins exposure to protease enzymes;protease XIV, protease XXIII and α-chymotrypsin type II were studied in this work. The degradation b...The degradation behaviors of Thai Bombyx mori called Samrong and Nanglai silk fibroins exposure to protease enzymes;protease XIV, protease XXIII and α-chymotrypsin type II were studied in this work. The degradation behaviors were expressed by their weight loss, morphological and secondary structure changes as well as thermal properties. Samrong showed higher percentage of weight loss than Nanglai. SEM micrographs indicated that silk fibroin were de- stroyed and showed many holes on their fiber surfaces. All of silk samples were increasable destroyed when exposure to the protease enzyme for long incubation period. With thermal analysis, both silk fibroin presented the thermal stability in the same profile. The result suggested that the selected silk fibroin should be composed of similar pattern of amino acids and their ratios. However, the protease susceptibility of each silk fibroin slightly varied in case of morphology observation. This might be affected by their genetic variety.展开更多
Fungi are the key agents in litter decomposition in forest ecosystems. However, the specific roles of the interactions between different fungal species during litter decomposition process are unclear. To evaluate the ...Fungi are the key agents in litter decomposition in forest ecosystems. However, the specific roles of the interactions between different fungal species during litter decomposition process are unclear. To evaluate the interactions, two fungi strains with significantly different morphs were isolated from the soils of Quercus acutissima forest and Pinus massoniana forest, and inoculated in the litter powder of Quercus acutissima leaves and Pinus massoniana needles with grown separately and in coexistence equally through a microcosm experiment. The enzyme activities were determined as a proxy for microbial activities. The results showed that the degradative enzymes involved in litter decomposition showed varying dynamics pattern during the incubation period. The interactions between the two fungi strains are synergism, and benefit to each other according to enzyme activities, suggesting that a fungi strain growth was accelerated by the presence of other fungi strain during litter decomposition process. However, the interactions of the two fungi strains were bilateral antagonism inoculated in the litter powder of Quercus acutissima leaves according to cellobiohydrolase activities. The synergism, despite bilateral antagonism in an exceptional case, may be an important factor controlling the fungal colonization and growth on litter substrate. The results implied that more fungal species may accelerate litter decomposition rates due to their mutual cooperation.展开更多
A fungus(WR-C1) decomposed cellulose was isolated from a hypothermal litter layer using Congo red medium as the preliminary screening culture medium and then using a filter as the secondary screening medium at low t...A fungus(WR-C1) decomposed cellulose was isolated from a hypothermal litter layer using Congo red medium as the preliminary screening culture medium and then using a filter as the secondary screening medium at low temperature. The experiment showed that the weight loss rate of filter paper on the 15 th days could reach 30.69%. A morphologic and ITS gene sequence analysis suggested that CF-C1 was Cladosporium. We mainly studied the effects of culture time, inoculation amount, initial p H and different sources of carbon, nitrogen and inorganic salt on the cellulase production of strain WR-C1. Under optimum cultural condition, the highest value of WR-C1 enzyme production and filter paper enzyme were 3.27 U · m L~(-1) and 0.51 U · m L~(-1).展开更多
With the rapid increase of lubricant consumption, oil contamination becomes more serious. Biotreatment is an important method to remove oil contamination with some advantages. In this study, acclimatized oil- contamin...With the rapid increase of lubricant consumption, oil contamination becomes more serious. Biotreatment is an important method to remove oil contamination with some advantages. In this study, acclimatized oil- contaminated soil and used lubricating oil were sampled to isolate lubricant-degrading strains by several methods. 51 isolates were obtained and 24-well plates were employed to assess bacterial potential in high- throughput screening. The method was noted for the prominence of oil-water two-phase system with saving chemicals, shortening cycles and lessening workloads. In order to decrease inaccuracy, subculture and resting cells were inoculated into mineral salt medium with 200 μ1 oil in well plates for the cultivation at 37 ℃ for 5 and 7 days, and the biodegradation potential was characterized by the changes of oil film and cell density. With appropriate evaluation by shaking flask tests, 5 isolates were retained for their potentials with the maxi- mum biodegradation from 1500 to 2200 mg· L-1 and identified as Acidovorax dtrulli, Pseudomonos balearica, Adnetobacterjohnsonii (two isolates with different biodegradation potentials) and Addovorax avenae using 16S rRNA sequencing analysis. Also, lipase activity was determined using indicator titration and p-nitrophenyl palmitate (p-NPP) methods. The results indicated that only p-NPP was successful to test lipase activity with the range of 1.93-6.29 mg· L-1 Although these five strains could degrade 1000 mg· L-1 lubricating oil in 158-168 h, there existed distinct difference in enzyme activity, which demonstrates that lipase activity could not be used as the criterion to evaluate microbial biodegradation potential for petroleum hydrocarbons.展开更多
Changes of and correlation among root tolerance index (RTI), root Aluminum (Al) content, root/shoot ratio (RSR), root malondialdehyde (MDA) content, and Superoxide dismutase (SOD) and peroxidase (POD) isoforms of maiz...Changes of and correlation among root tolerance index (RTI), root Aluminum (Al) content, root/shoot ratio (RSR), root malondialdehyde (MDA) content, and Superoxide dismutase (SOD) and peroxidase (POD) isoforms of maize YQ 7-96 were investigated under Al stress and removal of the stress (RS). Consequently, Al stress led to significant decreases in RTI, RSR, SOD and POD activities, but resulted in significant increase in root MD A and, Al accumulation in the tissues;Root SOD and POD activities did not correlate with Al and MDA contents in roots;The activities of SOD and POD were much lower in roots than in leaves. It can be concluded that (1) Al stress can lead to lipid peroxidation;(2) there is a larger POD family composed of different POD isoforms, some of which are of tissue-specific expression and play different roles in detoxification of Al in maize;(3) for POD isoforms, POD 2 is root-specific. POD 6 and POD 7 are all leaf-specific, POD 5 is not only root-specific but also RS-responsive;(4) high sensitivity of maize to Al is in part associated with much lower activities of both SOD and POD in roots;and (5) more importantly, both SOD and POD are therefore hinted to be not key players in prevention against Al-induced lipid peroxidation.展开更多
To clone and identify the gene encoding human ubiquitin binding enzym e 2 and study its expression pattern. Methods. According to the sequence of human EST, which is highly homologous to t he mouse ubiquitin binding/c...To clone and identify the gene encoding human ubiquitin binding enzym e 2 and study its expression pattern. Methods. According to the sequence of human EST, which is highly homologous to t he mouse ubiquitin binding/conjugating enzyme (E2), primers were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by bioinformati cs technique and its expression pattern was studied by using multiple tissue No rthern blot. Results. Two cDNA clones encoding human ubiquitin conjugating enzyme have been i solated and identified. Both containing the ubiquitin conjugating domain, the 2 cDNA clones are 88% identical in amino acid sequences and splicing isoforms to each other only with an exon excised to form the short sequence. They belong to a highly conserved and widely expressed E2 enzyme family. Northern blot shows th at they are expressed exclusively in adult human heart, placenta, and pancreas b ut no transcripts can be detected in brain, lung, liver, skeletal muscle or kidn ey. Conclusions. The gene encoding human ubiquitin binding enzyme is expressed under temporal control. As a key enzyme in the degradation of proteins, ubiquitin con jugating enzymes play a central role in the expression regulation on the level o f post translation.展开更多
[Objectives]To isolate and identify characteristics of the cellulose-degrading bacteria.[Methods]In view of the poor effect of crop straw composting,the strains which can degrade cellulose were isolated and purified f...[Objectives]To isolate and identify characteristics of the cellulose-degrading bacteria.[Methods]In view of the poor effect of crop straw composting,the strains which can degrade cellulose were isolated and purified from the soil piled with rice straw,and the strains A2 and A5 with high efficiency of cellulose degradation were finally screened through the transparent circle,cellulase activity and filter paper disintegration experiments.[Results]It was confirmed that the strains A2 and A5 could degrade cellulose well.The results of 16s rDNA showed that A2 was Pseudoxanthomonas mexicana and A5 was Bacillus cibi.[Conclusions]The results of this study are expected to provide high-quality strain resources for the degradation of cellulose in straw,and have important application value for improving the efficiency of straw composting.展开更多
In semi-arid areas of China,gravel and sand mulch is a farming technique with a long history.In this study,a sample survey was conducted on long term gravel sand mulch observational fields in the Northwest Loess Plate...In semi-arid areas of China,gravel and sand mulch is a farming technique with a long history.In this study,a sample survey was conducted on long term gravel sand mulch observational fields in the Northwest Loess Plateau to determine the effects of long term mulch on soil microbial and soil enzyme activities.We found that after long term gravel-sand mulch,compared with bare ground,soil organic matter,alkali nitrogen,conductivity decreased,while pH and soil moisture increased.Urease,saccharase and catalase decreased with increased mulch thickness,while alkaline phosphatase was reversed.The results of Illumina MiSeq sequencing shows that after gravel-sand mulch,the bacterial and fungal community structure was different from bare land,and the diversity was reduced.Compared with bare land,the bacteria Proteobacteria and Acidobacteria abundance increased with increased thickness,and Actinobacteria was opposite.Also,at the fungal genus level,Fusarium abundance was significantly reduced,and Remersonia was significantly increased,compared with bare land.Redundancy analysis(RDA)revealed that soil environmental factors were important drivers of bacterial community changes.Overall,this study revealed some of the reasons for soil degradation after long term gravel-sand mulch.Therefore,it is recommended that the addition of exogenous soil nutrients after long term gravel-sand can help improve soil quality.展开更多
基金supported by the National Natural Science Foundation of China,No.81303248,81603321the Natural Science Foundation of Heilongjiang Province of China,No.H2015028+1 种基金a grant from the Nursing Program for Young Scholars of Heilongjiang Province of China,No.UNPYSCT-2016116the Scientific Research Fund for Doctors of Qiqihar Medical University in China,No.QY2016B-09
文摘Kai Xin San is a Chinese herbal formula composed of Radix Ginseng, Poria, Radix Polygalae and Acorus Tatarinowii Rhizome. It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β (Aβ) induced cognitive dysfunction and is neuroprotective in vivo, but its precise mechanism remains unclear. Expression of insulin-degrading enzyme (IDE), which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 (200 μM, 5 μL) into the hippocampus and subsequently administered Kai Xin San (0.54 or 1.08 g/kg/d) intragastrically for 21 consecutive days. Hematoxylin eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.
基金the National Natural Science Foundation of China (31171877)the Youth Foundation Program for Science and Technology of Hebei Province, China (2010141)
文摘A strain of Bacillus subtilis strain YB 1, isolated and preserved in our lab., showed a high nicosulfuron-degrading activity. Optimization of culture conditions on production of nicosulfuron-degrading enzyme from Bacillus subtilis strain YB 1 was carried out through mono-factor experiments. The characterization of degrading enzyme(s) was studied in this paper. The results showed that B. subtilis YB1 can use nicosulfuron as sole carbon source under aerobic condition. The key enzyme(s) involved in the initial biodegradation of nicosulfuron was localized to extracellular proteins and showed to be induced expressed. Enzyme-specific activity was up to 89.34 U mg-1 at pH 8.0 and 30℃, incubation for 96 h, inoculum 4.5x108 CFU mL-1 in Luria-Bertani liquid medium with nicosulfuron of 40 mg L-1. The maximum degradation rate of extracellular crude enzymes on nicosulfuron was 66% at pH 9.0, 35℃ in the enzymatic reaction system with nicosulfuron of 5 mg L-1. This degrading enzyme(s) was sensitive to high temperature, but kept high activity under alkaline conditions.
基金supported by the National Key Tech-nology R&D Program of China (2008BADA6B05)the National Natural Science Foundation of China(30871714)
文摘This study was designed to analyze the changes of phytohormone concentrations, calcium fraction, and the activities of degrading enzymes during calcium-inhibited and ethyleneglycol-bis-(β-aminoethyl ether)N, N'-tetraacetic acid (EGTA)-induced abscission of tomato pedicel explants. Added calcium caused an increase in the total calcium content within the abscission zone and produced a corresponding reduction (20%) in pedicel explant abscission. As expected, EGTA treatment produced the opposite effect and resulted in a decrease in the total calcium content, while accelerating abscission of pedicel explants. Hormone analysis revealed that indole-3-acetic acid (IAA) concentrations in the abscission zone first decreased and then increased before the occurrence of abscission in all treatments, with the greatest effect produced by addition of EGTA. Similarly, abscisic acid (ABA), and gibberellin (GA1+3) concentrations, and ethylene production were elevated in the abscission zone during the first 16 h before abscission when explants imbibed EGTA. With calcium treatment, the concentrations of ABA, ethylene, and GA1+3 also increased in pedicels throughout the first 16 h following exposure, but the increase was slower and less dramatic than with EGTA. Both cellulase and polygalacturonase were induced in the explants during abscission and the activities were also strengthened by treatment with EGTA. Calciumtreated explants produced lower hydrolysing enzyme activities than controls throughout abscission. Calcium played a role of mediating hormone balance and degrading enzymes activities and affected on abscission.
基金Supported by the Special Fund for Agro-scientific Research(3-44-2)the Special Fund for Basic Research of Central Universities in the Public Interest,China(N0.2009hzs1J011,NO.2011hzs1J027,NO.2011hzs1J004)
文摘[ Objective ] This study aimed to confirm the roles of cell wall degrading enzymes (CWDEs) produced by Botryodiplodia theobromae Pat. in the infec- tion of mango fruits. [ Method] Change of activities of five types of CWDEs produced by B. theobromae Pat. were studied under both in vitro culture and inocula- tion conditions, along with the pathogenicity and the ability of producing CWDEs of four post-harvest fangal pathogens(B, theobromae Pat. , Colletotrichum gloeos- porioides Penz. , Phomopsis mangiferae Ahmad and Dothiorella dominicana Pet. et Cif. ) which cause stem-end rot of mangoes. [ Result] B. theobromae Pat. was a- ble to produce polygalacturonase(PG), pectinmethylgalacturonase(PMG), polygalacturonic acid trans-eliminase (PGTE), pectin methyltrans-eliminase (PMTE) and cellulase ( Cx. ) under both in vitro culture and inoculation conditions, of which activities of PG, Cx and PMG were significantly higher in than that in either PGTE or PMTE. Among three primary CWDEs, the peak of activities of PG and Cx appeared earlier and that of PMG occured later. The pathogenicity of B. theo- bromae Pat. was significantly higher than that of any other three pathogens; it is the same with the abilities of producing pectinase. [ Conclusion] This paper pro- vides theoretical bases for further exploring the mechanism of host-pathogen interaction and decreasing the post-harvest loss of mango fruits.
文摘Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE). Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderma. Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique. Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities. Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.
基金Supported by Yantai Municipal Science and Technology Development Plan(2013ZH097)Scientific and Technological Innovation Fund for Students in Binzhou Medical University(BY2013DKCX122)
文摘[ Objective ] This study aimed to obtain the recombinant Pichia yeast strain which can efficiently degrade guar gum. The properties of the recombinant enzyme were studied preliminarily. [ Method ] A positive clone that could hydrolyze guar gum was obtained through the construction and functional screening of a soil genome library. Sequence analysis indicated that the 1485-bp clone encodes a 494-amino acid protein with a relative molecular mass of 53 949 kD, containing a cellulose-binding domain. The recombinant plasmid pHBM731 was generated by inserting the optimized target gene into a Pichia pastoris expression vector pHBMg05 that was transformed into three Pichia pastoris strains, GS115, KM71 and SMD1168. The biochemical properties of the enzyme were assessed. [ Result] The cloned galactonumnan (GM)-degrading enzyme was expressed and secreted by Pichia pastoris GSll5. High cell density fermentation was induced in recombi- nant Pichia pastoris at 25 and 28 ~C ; a higher enzyme activity was observed at an induction temperature of 28 ~C. The optimal temperature for the recombinant en- zyme is 60 ~C, and the optimal pH is 6.6. The enzyme activity was 38.61 U under optimal conditions. Over 50% of the enzyme activity was maintained under the optimal conditions after 9 h. Under the optimal conditions, the effect of metal ions on enzyme activity was analyzed. Ca2 + , Fe2 + and Li ~ slightly enhanced enzyme activity, while Mn2+ and Co2+ had little effect. Enzyme activity was modestly suppressed by Mg2~ , K~ and Na+ , but considerably suppressed by Ag2~ and Zn2~ , with Cu2 + showing the strongest inhibitory effects. [ Conclusion] A novel GM-degrading enzyme expressed by soil yeast was cloned, which can potentially be used in industrial applications to obtain eommereially useful guar gum-degradation products.
文摘Peanuts can be affected by the presence of pathogenic microorganisms. The fungus <i>Thecaphora frezii</i> (<i>T. frezii</i>), which belongs to the taxonomic class Ustilaginomycetes, is the causal agent of the disease known as “peanut smut”. The life cycle of this fungus includes three stages, namely teliospores, basidiospores and hyphae. In the hyphae stage, infection occurs in the peanut plant, which requires the involvement of some enzymes secreted by the fungus. These include the Plant Cell Wall-Degrading Enzymes (PCWDEs), which degrade various polysaccharides. This study aimed to identify the presence of transcript for enzymes belonging to the PCWDEs from three stages of <i>T. frezii</i>. For this, total RNA was extracted from the three ontogenetic stages of <i>T. frezii</i>. These samples were analyzed using an RNA-Seq approach and some transcripts were quantified using Real Time PCR. The analysis of the data provided by the RNA-Seq of the three <i>T. frezii</i> stages, it was possible to identify some transcripts that could encode enzymes compatible with polysaccharides degradation that are part of the plant cell wall. In <i>T. frezii</i> transcriptome, 40 deduced proteins would be enzymes with functions of PCWDEs were identified. They were divided into 27 glycoside hydrolases;two polysaccharide lyases;three carbohydrate esterases and eight enzymes with auxiliary activities. In addition, the fungal SNF1 gene was identified whose activity could be affected by high glucose level, and indirectly influence the levels of some PCWDEs. The analysis of the PCWDEs could help to understand part of the fungal infection process and possibly find substances that can control its development.
基金supported by the National Natural Science Foundation of China(30870419,40971151)Strategic Priority Research Program Climate Change:Carbon Budget and Related Issues of the Chinese Academy of Sciences(XDA05050204)
文摘We used a litterbag method to investigate litter decomposition and related soil degradative enzyme activities across four seasons in a broad-leaved forest and a coniferous forest on Zijin Mountain in sub-tropical China. Across four seasons, we quantified litter mass losses, soil pH values, and related soil degradative enzyme activities. Litter decomposition rates differed significantly by season. Litter decomposi- tion rates of broadleaf forest leaves were higher than for coniferous for- ests needles across four seasons, and maximal differences in litter de- composition rates between the two litter types were found in spring.
基金supported by the National Key Basic Research Program of China (2013CB127105)the National Natural Science Foundation of China (30671466)+1 种基金China Litchi and Logan Research System (CARS-33-14)Guangdong Fruit Research System,China (2009-356)
文摘Heat and acid treatments were reported to be a promising substitute for SO2 fumigation in color protection of postharvest lychee (Litchi chinensis) fruits, but the mechanism was not clear. In the present study, hot water (70℃) dipping followed by immersion in 2% HC1 (heat-acid) substantially protected the red color of the fruit during storage at 25℃ and inhibited anthocyanin degradation while hot water dipping alone (heat) led to rapidly browning and about 90% loss in anthocyanin content. The pH values in the pericarp of the heat-acid treated fruit dropped to 3.2, while the values maintained around 5.0 in the heat-treated and control fruit. No significantly different pH values were detected among the arils of heat-acid, heat treated and control fruit. Heat-acid treatment dramatically reduced the activities of anthocyanin degradation enzyme (ADE), peroxidase (POD) and polyphenol oxidase in the pericarp. A marked reduction in LcPOD gene expression was also detected in heat-acid treated fruit, in contrast, induction was found in heat treated fruit. The pericarp of heat-acid treated fruit exhibited significantly lower respiration rate but faster water loss than that of the untreated or heat treated fruit. Taken together, heat treatment triggered quick browning and anthocyanin loss in lychee fruit, while heat-acid treatment protected the fruit color by a great reduction in the activities/gene expression of anthocyanin degradation enzymes and acidification of lychee pericarp.
基金supported by the Key Project of State Tobacco Monoply Administration of China(3300806156)
文摘The mechanism of carotenoid degradation and the changes in the activities of related enzymes in flue-cured tobacco at the leaf-drying stage during the bulk-curing process were studied in order to provide theoretical basis for optimization of curing technology. The effect of different rising speeds of temperature on the carotenoid degradation and the related enzymes activities at the color-fixing stage during the bulk curing process was studied by using the electric-heated fluecuring barn designed by Henan Agricultural University, China, based on curing technology with yellowing at low temperature and moderate humidity and leaf drying at moderate humidity. The results showed that the carotenoid degradation components (β-carotene, lutein, neoxanthin, and violaxthin) decreased gradually at the color-fixing stage during the bulk curing process. The carotenoid degradation components viz.,β-carotene, lutein, neoxanthin, and violaxthin at the slow heating curing (T1) were relatively higher than the rapid heating curing (T2) accounting for 10, 2, 32 and 32% respectively, but there were no differences among treatments (P〉 0.05). The effect of different conditions of curing on the activities of enzymes related to carotenoids degradation were significant. The lipoxygenase, phenylalanine ammonialyase, peroxidase, and polyphenol oxidase enzymes had a bidirectional effect on the quality of tobacco leaves and it was beneficial to form more premise matter of aroma based on the higher enzyme activities at the early leaf-drying stage. The slow heating could regulate the change in various enzymes' activities reasonably, making cell redox reaction to reach the dynamic balance and make the degradation of carotenoids adequately. Meanwhile, it could avoid the occurrence of browning reaction and provide foundation for improving the quality of tobacco and optimization of technology for bulk curing and further enhancing aroma.
文摘The degradation behaviors of Thai Bombyx mori called Samrong and Nanglai silk fibroins exposure to protease enzymes;protease XIV, protease XXIII and α-chymotrypsin type II were studied in this work. The degradation behaviors were expressed by their weight loss, morphological and secondary structure changes as well as thermal properties. Samrong showed higher percentage of weight loss than Nanglai. SEM micrographs indicated that silk fibroin were de- stroyed and showed many holes on their fiber surfaces. All of silk samples were increasable destroyed when exposure to the protease enzyme for long incubation period. With thermal analysis, both silk fibroin presented the thermal stability in the same profile. The result suggested that the selected silk fibroin should be composed of similar pattern of amino acids and their ratios. However, the protease susceptibility of each silk fibroin slightly varied in case of morphology observation. This might be affected by their genetic variety.
文摘Fungi are the key agents in litter decomposition in forest ecosystems. However, the specific roles of the interactions between different fungal species during litter decomposition process are unclear. To evaluate the interactions, two fungi strains with significantly different morphs were isolated from the soils of Quercus acutissima forest and Pinus massoniana forest, and inoculated in the litter powder of Quercus acutissima leaves and Pinus massoniana needles with grown separately and in coexistence equally through a microcosm experiment. The enzyme activities were determined as a proxy for microbial activities. The results showed that the degradative enzymes involved in litter decomposition showed varying dynamics pattern during the incubation period. The interactions between the two fungi strains are synergism, and benefit to each other according to enzyme activities, suggesting that a fungi strain growth was accelerated by the presence of other fungi strain during litter decomposition process. However, the interactions of the two fungi strains were bilateral antagonism inoculated in the litter powder of Quercus acutissima leaves according to cellobiohydrolase activities. The synergism, despite bilateral antagonism in an exceptional case, may be an important factor controlling the fungal colonization and growth on litter substrate. The results implied that more fungal species may accelerate litter decomposition rates due to their mutual cooperation.
基金Supported by the Post-doctoral Research Starting Fund of Heilongjiang Province(LBH-Q13020)
文摘A fungus(WR-C1) decomposed cellulose was isolated from a hypothermal litter layer using Congo red medium as the preliminary screening culture medium and then using a filter as the secondary screening medium at low temperature. The experiment showed that the weight loss rate of filter paper on the 15 th days could reach 30.69%. A morphologic and ITS gene sequence analysis suggested that CF-C1 was Cladosporium. We mainly studied the effects of culture time, inoculation amount, initial p H and different sources of carbon, nitrogen and inorganic salt on the cellulase production of strain WR-C1. Under optimum cultural condition, the highest value of WR-C1 enzyme production and filter paper enzyme were 3.27 U · m L~(-1) and 0.51 U · m L~(-1).
基金Supported by the National Natural Science Foundation of China(21376285)Chongqing Natural Science Foundation(CSTC2013jcyj A20014)+3 种基金Open Funding Project of the Key Laboratory of Systems BioengineeringMinistry of Educationand Scientific Platform ProjectMinistry of Education(FYKF201506)
文摘With the rapid increase of lubricant consumption, oil contamination becomes more serious. Biotreatment is an important method to remove oil contamination with some advantages. In this study, acclimatized oil- contaminated soil and used lubricating oil were sampled to isolate lubricant-degrading strains by several methods. 51 isolates were obtained and 24-well plates were employed to assess bacterial potential in high- throughput screening. The method was noted for the prominence of oil-water two-phase system with saving chemicals, shortening cycles and lessening workloads. In order to decrease inaccuracy, subculture and resting cells were inoculated into mineral salt medium with 200 μ1 oil in well plates for the cultivation at 37 ℃ for 5 and 7 days, and the biodegradation potential was characterized by the changes of oil film and cell density. With appropriate evaluation by shaking flask tests, 5 isolates were retained for their potentials with the maxi- mum biodegradation from 1500 to 2200 mg· L-1 and identified as Acidovorax dtrulli, Pseudomonos balearica, Adnetobacterjohnsonii (two isolates with different biodegradation potentials) and Addovorax avenae using 16S rRNA sequencing analysis. Also, lipase activity was determined using indicator titration and p-nitrophenyl palmitate (p-NPP) methods. The results indicated that only p-NPP was successful to test lipase activity with the range of 1.93-6.29 mg· L-1 Although these five strains could degrade 1000 mg· L-1 lubricating oil in 158-168 h, there existed distinct difference in enzyme activity, which demonstrates that lipase activity could not be used as the criterion to evaluate microbial biodegradation potential for petroleum hydrocarbons.
文摘Changes of and correlation among root tolerance index (RTI), root Aluminum (Al) content, root/shoot ratio (RSR), root malondialdehyde (MDA) content, and Superoxide dismutase (SOD) and peroxidase (POD) isoforms of maize YQ 7-96 were investigated under Al stress and removal of the stress (RS). Consequently, Al stress led to significant decreases in RTI, RSR, SOD and POD activities, but resulted in significant increase in root MD A and, Al accumulation in the tissues;Root SOD and POD activities did not correlate with Al and MDA contents in roots;The activities of SOD and POD were much lower in roots than in leaves. It can be concluded that (1) Al stress can lead to lipid peroxidation;(2) there is a larger POD family composed of different POD isoforms, some of which are of tissue-specific expression and play different roles in detoxification of Al in maize;(3) for POD isoforms, POD 2 is root-specific. POD 6 and POD 7 are all leaf-specific, POD 5 is not only root-specific but also RS-responsive;(4) high sensitivity of maize to Al is in part associated with much lower activities of both SOD and POD in roots;and (5) more importantly, both SOD and POD are therefore hinted to be not key players in prevention against Al-induced lipid peroxidation.
文摘To clone and identify the gene encoding human ubiquitin binding enzym e 2 and study its expression pattern. Methods. According to the sequence of human EST, which is highly homologous to t he mouse ubiquitin binding/conjugating enzyme (E2), primers were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by bioinformati cs technique and its expression pattern was studied by using multiple tissue No rthern blot. Results. Two cDNA clones encoding human ubiquitin conjugating enzyme have been i solated and identified. Both containing the ubiquitin conjugating domain, the 2 cDNA clones are 88% identical in amino acid sequences and splicing isoforms to each other only with an exon excised to form the short sequence. They belong to a highly conserved and widely expressed E2 enzyme family. Northern blot shows th at they are expressed exclusively in adult human heart, placenta, and pancreas b ut no transcripts can be detected in brain, lung, liver, skeletal muscle or kidn ey. Conclusions. The gene encoding human ubiquitin binding enzyme is expressed under temporal control. As a key enzyme in the degradation of proteins, ubiquitin con jugating enzymes play a central role in the expression regulation on the level o f post translation.
基金Supported by Innovation and Entrepreneurship Training Program for College Students (202310580018).
文摘[Objectives]To isolate and identify characteristics of the cellulose-degrading bacteria.[Methods]In view of the poor effect of crop straw composting,the strains which can degrade cellulose were isolated and purified from the soil piled with rice straw,and the strains A2 and A5 with high efficiency of cellulose degradation were finally screened through the transparent circle,cellulase activity and filter paper disintegration experiments.[Results]It was confirmed that the strains A2 and A5 could degrade cellulose well.The results of 16s rDNA showed that A2 was Pseudoxanthomonas mexicana and A5 was Bacillus cibi.[Conclusions]The results of this study are expected to provide high-quality strain resources for the degradation of cellulose in straw,and have important application value for improving the efficiency of straw composting.
基金This study was funded by the National Key R&D Program(Grant No.2016YFC0501403-3).
文摘In semi-arid areas of China,gravel and sand mulch is a farming technique with a long history.In this study,a sample survey was conducted on long term gravel sand mulch observational fields in the Northwest Loess Plateau to determine the effects of long term mulch on soil microbial and soil enzyme activities.We found that after long term gravel-sand mulch,compared with bare ground,soil organic matter,alkali nitrogen,conductivity decreased,while pH and soil moisture increased.Urease,saccharase and catalase decreased with increased mulch thickness,while alkaline phosphatase was reversed.The results of Illumina MiSeq sequencing shows that after gravel-sand mulch,the bacterial and fungal community structure was different from bare land,and the diversity was reduced.Compared with bare land,the bacteria Proteobacteria and Acidobacteria abundance increased with increased thickness,and Actinobacteria was opposite.Also,at the fungal genus level,Fusarium abundance was significantly reduced,and Remersonia was significantly increased,compared with bare land.Redundancy analysis(RDA)revealed that soil environmental factors were important drivers of bacterial community changes.Overall,this study revealed some of the reasons for soil degradation after long term gravel-sand mulch.Therefore,it is recommended that the addition of exogenous soil nutrients after long term gravel-sand can help improve soil quality.